Yeast-like ascomycetous fungi of the family Saccharomycetaceae, order SACCHAROMYCETALES isolated from exuded tree sap.
A colorless, flammable liquid used in the manufacture of FORMALDEHYDE and ACETIC ACID, in chemical synthesis, antifreeze, and as a solvent. Ingestion of methanol is toxic and may cause blindness.
Proteins prepared by recombinant DNA technology.
Protein factors released from one species of YEAST that are selectively toxic to another species of yeast.
Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.
The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells.
An order of fungi in the phylum Ascomycota that multiply by budding. They include the telomorphic ascomycetous yeasts which are found in a very wide range of habitats.
A telecommunication system combining the transmission of a document scanned at a transmitter, its reconstruction at a receiving station, and its duplication there by a copier.
The sounds produced by humans by the passage of air through the LARYNX and over the VOCAL CORDS, and then modified by the resonance organs, the NASOPHARYNX, and the MOUTH.
An infectious disease clinically similar to epidemic louse-borne typhus (TYPHUS, EPIDEMIC LOUSE-BORNE), but caused by RICKETTSIA TYPHI, which is transmitted from rat to man by the rat flea, XENOPSYLLA CHEOPIS.
Water in its gaseous state. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed)
An enzyme that plays a role in the PENTOSES and GLUCURONATES interconversion pathway by catalyzing the oxidation of XYLITOL to D-xylulose. This enzyme has been found to be specific for NAD+.
Anaerobic degradation of GLUCOSE or other organic nutrients to gain energy in the form of ATP. End products vary depending on organisms, substrates, and enzymatic pathways. Common fermentation products include ETHANOL and LACTIC ACID.
Inorganic and organic derivatives of sulfuric acid (H2SO4). The salts and esters of sulfuric acid are known as SULFATES and SULFURIC ACID ESTERS respectively.
Reversibly catalyzes the oxidation of a hydroxyl group of sugar alcohols to form a keto sugar, aldehyde or lactone. Any acceptor except molecular oxygen is permitted. Includes EC 1.1.1.; EC 1.1.2. and EC 1.1.99.
An iron-binding beta1-globulin that is synthesized in the LIVER and secreted into the blood. It plays a central role in the transport of IRON throughout the circulation. A variety of transferrin isoforms exist in humans, including some that are considered markers for specific disease states.
Membrane glycoproteins found in high concentrations on iron-utilizing cells. They specifically bind iron-bearing transferrin, are endocytosed with its ligand and then returned to the cell surface where transferrin without its iron is released.
The study of crystal structure using X-RAY DIFFRACTION techniques. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)
A metallic element with atomic symbol Fe, atomic number 26, and atomic weight 55.85. It is an essential constituent of HEMOGLOBINS; CYTOCHROMES; and IRON-BINDING PROTEINS. It plays a role in cellular redox reactions and in the transport of OXYGEN.
A glycoprotein albumin from hen's egg white with strong iron-binding affinity.
Models used experimentally or theoretically to study molecular shape, electronic properties, or interactions; includes analogous molecules, computer-generated graphics, and mechanical structures.
An enzyme of the hydrolase class that catalyzes the reaction of triacylglycerol and water to yield diacylglycerol and a fatty acid anion. It is produced by glands on the tongue and by the pancreas and initiates the digestion of dietary fats. (From Dorland, 27th ed) EC 3.1.1.3.
An antineoplastic agent that is a derivative of progesterone and used to treat advanced breast cancer.
A genus of mitosporic fungi containing about 100 species and eleven different teleomorphs in the family Trichocomaceae.
An enzyme of the hydrolase class that catalyzes the reaction of triacylglycerol and water to yield diacylglycerol and a fatty acid anion. The enzyme hydrolyzes triacylglycerols in chylomicrons, very-low-density lipoproteins, low-density lipoproteins, and diacylglycerols. It occurs on capillary endothelial surfaces, especially in mammary, muscle, and adipose tissue. Genetic deficiency of the enzyme causes familial hyperlipoproteinemia Type I. (Dorland, 27th ed) EC 3.1.1.34.
The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION.
A species of imperfect fungi from which the antibiotic nidulin is obtained. Its teleomorph is Emericella nidulans.
The mallow family of the order Malvales, subclass Dilleniidae, class Magnoliopsida. Members include GOSSYPIUM, okra (ABELMOSCHUS), HIBISCUS, and CACAO. The common names of hollyhock and mallow are used for several genera of Malvaceae.
DNA molecules capable of autonomous replication within a host cell and into which other DNA sequences can be inserted and thus amplified. Many are derived from PLASMIDS; BACTERIOPHAGES; or VIRUSES. They are used for transporting foreign genes into recipient cells. Genetic vectors possess a functional replicator site and contain GENETIC MARKERS to facilitate their selective recognition.
DNA sequences which are recognized (directly or indirectly) and bound by a DNA-dependent RNA polymerase during the initiation of transcription. Highly conserved sequences within the promoter include the Pribnow box in bacteria and the TATA BOX in eukaryotes.

Kodamaea nitidulidarum, Candida restingae and Kodamaea anthophila, three new related yeast species from ephemeral flowers. (1/1453)

Three new yeast species were discovered during studies of yeasts associated with ephemeral flowers in Brazil, Australia and Hawaii. Their physiological and morphological similarity to Kodamaea (Pichia) ohmeri suggested a possible relationship to that species, which was confirmed by rDNA sequencing. Kodamaea nitidulidarum and Candida restingae were found in cactus flowers and associated nitidulid beetles in sand dune ecosystems (restinga) of South-eastern Brazil. Over 350 strains of Kodamaea anthophila were isolated from Hibiscus and morning glory flowers (Ipomoea spp.) in Australia, and from associated nitidulid beetles and Drosophila hibisci. A single isolate came from a beach morning glory in Hawaii. Expansion of the genus Kodamaea to three species modified the existing definition of the genus only slightly. The type and isotype strains are as follows: K. nitidulidarum strains UFMG96-272T (h+; CBS 8491T) and UFMG96-394I (h-; CBS 8492I); Candida restingae UFMG96-276T (CBS 8493T); K. anthophila strains UWO(PS)95-602.1T (h+; CBS 8494T), UWO(PS)91-893.2I (h-; CBS 8495I) and UWO(PS)95-725.1I (h-; CBS 8496I).  (+info)

Glycosylation of asparagine-28 of recombinant staphylokinase with high-mannose-type oligosaccharides results in a protein with highly attenuated plasminogen activator activity. (2/1453)

The properties of recombinant staphylokinase (SakSTAR) expressed in Pichia pastoris cells have been determined. The single consensus N-linked oligosaccharide linkage site in SakSTAR (at Asn28 of the mature protein) was occupied in approximately 50% of the expressed protein with high-mannose-type oligosaccharides. The majority of these glycans ranged in polymerization state from Man8GlcNAc2 to Man14GlcNAc2, with the predominant species being Man10GlcNAc2 and Man11GlcNAc2. Glycosylated SakSTAR (SakSTARg) did not differ from its aglycosyl form in its aggregation state in solution, its thermal denaturation properties, its ability to form a complex with human plasmin (hPm), the amidolytic properties of the respective SakSTAR-hPm complexes, or its ability to liberate the amino-terminal decapeptide required for formation of a functional SakSTAR-hPm plasminogen activator complex. However, this latter complex with SakSTARg showed a greatly reduced ability to activate human plasminogen (hPg) as compared with the same complex with the aglycosyl form of SakSTAR. We conclude that glycosylation at Asn28 does not affect the structural properties of SakSTAR or its ability to participate in the formation of an active enzymatic complex with hPm, but it is detrimental to the ability of the SakSTAR-hPm complex to serve as a hPg activator. This is likely due to restricted access of hPg to the active site of the SakSTARg-hPm complex.  (+info)

Protection of mice against a lethal influenza virus challenge after immunization with yeast-derived secreted influenza virus hemagglutinin. (3/1453)

The A/Victoria/3/75 (H3N2-subtype) hemagglutinin (HA) gene was engineered for expression in Pichia pastoris as a soluble secreted molecule. The HA cDNA lacking the C-terminal transmembrane anchor-coding sequence was fused to the Saccharomyces cerevisiae alpha-mating factor secretion signal and placed under control of the methanol-inducible P. pastoris alcohol oxidase 1 (AOX1) promoter. Growth of transformants on methanol-containing medium resulted in the secretion of recombinant non-cleaved soluble hemagglutinin (HA0s). Remarkably, the pH of the induction medium had an important effect on the expression level, the highest level being obtained at pH 8.0. The gel filtration profile and the reactivity against a panel of different HA-conformation specific monoclonal antibodies indicated that HA0s was monomeric. Analysis of the N-linked glycans revealed a typical P. pastoris type of glycosylation, consisting of glycans with 10-12 glycosyl residues. Mice immunized with purified soluble hemagglutinin (HA0s) showed complete protection against a challenge with 10 LD50 of mouse-adapted homologous virus (X47), whereas all control mice succumbed. Heterologous challenge with X31 virus [A/Aichi/2/68 (H3N2-subtype)], resulted in significantly higher survival rates in the immunized group compared with the control group. These results, together with the safety, reliability and economic potential of P. pastoris, as well as the flexibility and fast adaptation of the expression system may allow development of an effective recombinant influenza vaccine.  (+info)

Cloning, mutagenesis, and structural analysis of human pancreatic alpha-amylase expressed in Pichia pastoris. (4/1453)

Human pancreatic alpha-amylase (HPA) was expressed in the methylotrophic yeast Pichia pastoris and two mutants (D197A and D197N) of a completely conserved active site carboxylic acid were generated. All recombinant proteins were shown by electrospray ionization mass spectrometry (ESI-MS) to be glycosylated and the site of attachment was shown to be Asn461 by peptide mapping in conjunction with ESI-MS. Treatment of these proteins with endoglycosidase F demonstrated that they contained a single N-linked oligosaccharide and yielded a protein product with a single N-acetyl glucosamine (GlcNAc), which could be crystallized. Solution of the crystal structure to a resolution of 2.0 A confirmed the location of the glycosyl group as Asn461 and showed that the recombinant protein had essentially the same conformation as the native enzyme. The kinetic parameters of the glycosylated and deglycosylated wild-type proteins were the same while the k(cat)/Km values for D197A and D197N were 10(6)-10(7) times lower than the wild-type enzyme. The decreased k(cat)/Km values for the mutants confirm that D197 plays a crucial role in the hydrolytic activity of HPA, presumably as the catalytic nucleophile.  (+info)

The N-terminal CUB-epidermal growth factor module pair of human complement protease C1r binds Ca2+ with high affinity and mediates Ca2+-dependent interaction with C1s. (5/1453)

The Ca2+-dependent interaction between complement serine proteases C1r and C1s is mediated by their alpha regions, encompassing the major part of their N-terminal CUB-EGF-CUB (where EGF is epidermal growth factor) module array. In order to define the boundaries of the C1r domain(s) responsible for Ca2+ binding and Ca2+-dependent interaction with C1s and to assess the contribution of individual modules to these functions, the CUB, EGF, and CUB-EGF fragments were expressed in eucaryotic systems or synthesized chemically. Gel filtration studies, as well as measurements of intrinsic Tyr fluorescence, provided evidence that the CUB-EGF pair adopts a more compact conformation in the presence of Ca2+. Ca2+-dependent interaction of intact C1r with C1s was studied using surface plasmon resonance spectroscopy, yielding KD values of 10.9-29.7 nM. The C1r CUB-EGF pair bound immobilized C1s with a higher KD (1.5-1.8 microM), which decreased to 31.4 nM when CUB-EGF was used as the immobilized ligand and C1s was free. Half-maximal binding was obtained at comparable Ca2+ concentrations ranging from 5 microM with intact C1r to 10-16 microM for C1ralpha and CUB-EGF. The isolated CUB and EGF fragments or a CUB + EGF mixture did not bind C1s. These data demonstrate that the C1r CUB-EGF module pair (residues 1-175) is the minimal segment required for high affinity Ca2+ binding and Ca2+-dependent interaction with C1s and indicate that Ca2+ binding induces a more compact folding of the CUB-EGF pair.  (+info)

Cloning, expression, and properties of a nonneuronal secreted acetylcholinesterase from the parasitic nematode Nippostrongylus brasiliensis. (6/1453)

We have isolated a full-length cDNA encoding an acetylcholinesterase secreted by the nematode parasite Nippostrongylus brasiliensis. The predicted protein is truncated in comparison with acetylcholinesterases from other organisms such that the carboxyl terminus aligns closely to the end of the catalytic domain of the vertebrate enzymes. The residues in the catalytic triad are conserved, as are the six cysteines which form the three intramolecular disulfide bonds. Three of the fourteen aromatic residues which line the active site gorge in the Torpedo enzyme are substituted by nonaromatic residues, corresponding to Tyr-70 (Thr), Trp-279 (Asn), and Phe-288 (Met). High level expression was obtained via secretion from Pichia pastoris. The purified enzyme behaved as a monomeric hydrophilic species. Although of invertebrate origin and possessing the above substitutions in the active site gorge residues, the enzyme efficiently hydrolyzed acetylthiocholine and showed minimal activity against butyrylthiocholine. It displayed excess substrate inhibition with acetylthiocholine at concentrations over 2. 5 mM and was highly sensitive to both active site and "peripheral" site inhibitors. Northern blot analysis indicated a progressive increase in mRNA for AChE B in parasites isolated from 6 days postinfection.  (+info)

Positive selection of novel peroxisome biogenesis-defective mutants of the yeast Pichia pastoris. (7/1453)

We have developed two novel schemes for the direct selection of peroxisome-biogenesis-defective (pex) mutants of the methylotrophic yeast Pichia pastoris. Both schemes take advantage of our observation that methanol-induced pex mutants contain little or no alcohol oxidase (AOX) activity. AOX is a peroxisomal matrix enzyme that catalyzes the first step in the methanol-utilization pathway. One scheme utilizes allyl alcohol, a compound that is not toxic to cells but is oxidized by AOX to acrolein, a compound that is toxic. Exposure of mutagenized populations of AOX-induced cells to allyl alcohol selectively kills AOX-containing cells. However, pex mutants without AOX are able to grow. The second scheme utilizes a P. pastoris strain that is defective in formaldehyde dehydrogenase (FLD), a methanol pathway enzyme required to metabolize formaldehyde, the product of AOX. AOX-induced cells of fld1 strains are sensitive to methanol because of the accumulation of formaldehyde. However, fld1 pex mutants, with little active AOX, do not efficiently oxidize methanol to formaldehyde and therefore are not sensitive to methanol. Using these selections, new pex mutant alleles in previously identified PEX genes have been isolated along with mutants in three previously unidentified PEX groups.  (+info)

Physiological effects and adjuvanticity of recombinant brushtail possum TNF-alpha. (8/1453)

The present paper describes the physiological properties of recombinant possum TNF-alpha and an adjuvant effect on antibody responses to the model protein antigen, keyhole limpet haemocyanin (KLH). For these studies recombinant possum TNF-alpha was produced in the yeast Pichia pastoris. The recombinant cytokine was secreted into the culture medium and purified by gel filtration. Possum TNF-alpha produced in this expression system was N-glycosylated and bioactive in two different assays. In a murine fibroblast L929 cytotoxicity assay, the possum TNF-alpha had lower specific activity compared to human TNF-alpha, while in a possum-specific assay, possum TNF-alpha enhanced the proliferation of PHA-stimulated possum thymocytes and was more active than human TNF-alpha. The physiological effect of the recombinant possum TNF-alpha was investigated in groups of possums administered doses of 6, 30 or 150 micrograms of cytokine. For each dose, TNF-alpha caused profound effects on the numbers of circulating leucocytes characterized by a three-to-four-fold increase in neutrophil numbers at 6-24 h after injection and an initial sharp decrease in lymphocyte numbers. The efficacy of TNF-alpha as an immunological adjuvant was determined in possums administered KLH (125 micrograms) in an aqueous or Al(OH)3-based formulation with or without added recombinant TNF-alpha (150 micrograms). Serum antibody responses to KLH were monitored by ELISA. The TNF-alpha stimulated two-fold and four-fold increases in antibody levels in aqueous and Al(OH)3-based vaccine formulations, respectively. The strongest antibody responses were observed in the group of possums that received KLH formulated in Al(OH)3 with addition of TNF-alpha.  (+info)

... By Julia Cino PhD ... Introduction Over th...The production of a functional protein is intimately related to the ce... Pichia pastoris Expression System /...,High,Yield,Protein,Production,from,Pichia,pastoris,Yeast:,A,Protocol,for,Benchtop,Fermentation,biological,advanced biology technology,biology laboratory technology,biology device technology,latest biology technology
TY - JOUR. T1 - Improvement in the secretory expression of recombinant Candida rugosa lipase in Pichia pastoris. AU - Kuo, Ting Chun. AU - Shaw, Jei Fu. AU - Lee, Guan Chiun. PY - 2015/12/1. Y1 - 2015/12/1. N2 - The yeast Candida rugosa can secrete a mixture of lipase isoenzymes (Lips), which have been widely applied in industry. Eight Lip genes (LIP1 to LIP8) have been identified and are expressed in Pichia pastoris. However, the expression level was not sufficient for economical industrial application. In this study, two combined processes of antibiotic selection and low-temperature culture efficiently elicited a high-level secretion of recombinant Lip2 in P. pastoris. The LIP2 gene copy number of the Pichia transformants was increased by sequential selections at gradually increasing Zeocin concentrations. After the first selection at 500 μg/mL of Zeocin, three clones (500-clones) with 2.4-fold to 5.8-fold improvement in Lip2 secretion were identified from 105 survival clones through lipase ...
TY - JOUR. T1 - Expression of recombinant actin 5C from Drosophila in the methylotrophyc yeast Pichia pastoris. AU - Nevzglyadova, O. V.. AU - Artemov, A. V.. AU - Zenin, V. V.. AU - Verkhusha, V. V.. AU - Shavlovsky, M. M.. AU - Povarova, O. I.. AU - Stepanenko, O. V.. AU - Kuznetsova, I. M.. AU - Turoverov, K. K.. PY - 2007/6/1. Y1 - 2007/6/1. N2 - A system of expression for the foreign actin gene in yeast cells Pichia pastoris has been developed. As a target protein, the Drosophila cytoplasmic actin 5C, which has 90% homology to the β-actin of higher eukaryotes, was used. In the present work, in order to develop conditions for biosynthesis of the target protein in yeast cells and a purification procedure for the recombinant protein, a GFP-actin fusion protein containing green fluorescent protein (GFP) as a fusion tag was expressed and purified. The size and survival of P. pastoris cells producing recombinant protein were characterized and shown to depend on the accumulation of recombinant ...
Several months back, a couple of postings about the Invitrogen Pichia pastoris expression system appeared. A look at the archives failed to find any answers to the posts. Does anyone out there have any experience (good or bad) with this expression system? I will summarize and post if necessary. Thanks _____________________________________ Dean R. Appling Dept. of Chemistry & Biochemistry Univ. of Texas at Austin Austin, TX 78712-1167 (512) 471-5842 (voice) (512) 471-5849 (fax) appling at utbc01.cm.utexas.edu ...
Yeast cells represent an established bioreactor to produce recombinant proteins for subunit vaccine development. In addition, delivery of vaccine antigens directly within heat-inactivated yeast cells is attractive due to the adjuvancy provided by the yeast cell. In this study, Pichia pastoris yeast lysates carrying the nucleoprotein (N) from the measles vaccine virus were evaluated as a novel subunit vaccine platform to deliver the circumsporozoite surface antigen (CS) of Plasmodium. When expressed in Pichia pastoris yeast, the N protein auto-assembles into highly multimeric ribonucleoparticles (RNPs). The CS antigen from Plasmodium berghei (PbCS) was expressed in Pichia pastoris yeast in fusion with N, generating recombinant PbCS-carrying RNPs in the cytoplasm of yeast cells. When evaluated in mice after 3-5 weekly subcutaneous injections, yeast lysates containing N-PbCS RNPs elicited strong anti-PbCS humoral responses, which were PbCS-dose dependent and reached a plateau by the pre-challenge time
Yeast cells represent an established bioreactor to produce recombinant proteins for subunit vaccine development. In addition, delivery of vaccine antigens directly within heat-inactivated yeast cells is attractive due to the adjuvancy provided by the yeast cell. In this study, Pichia pastoris yeast lysates carrying the nucleoprotein (N) from the measles vaccine virus were evaluated as a novel subunit vaccine platform to deliver the circumsporozoite surface antigen (CS) of Plasmodium. When expressed in Pichia pastoris yeast, the N protein auto-assembles into highly multimeric ribonucleoparticles (RNPs). The CS antigen from Plasmodium berghei (PbCS) was expressed in Pichia pastoris yeast in fusion with N, generating recombinant PbCS-carrying RNPs in the cytoplasm of yeast cells. When evaluated in mice after 3-5 weekly subcutaneous injections, yeast lysates containing N-PbCS RNPs elicited strong anti-PbCS humoral responses, which were PbCS-dose dependent and reached a plateau by the pre-challenge time
To help the researchers and pharmaceutical partners with discovering the most effective therapeutic glycoprotein, Creative Biolabs provides solutions on glycosylation, the most widely applied posttranslational modification (PTM) approach to change protein function and measure the recombinant biopharmaceutical immunogenicity. Based upon the efficient glycoengineered expression platforms (glyco-engineered mammalian cell expression system, glyco-engineered pichia pastoris expression system, and glyco-engineered plant-based expression system), Creative Biolabs is fully competent to handle the requests of therapeutic glycoprotein development and glycoengineering of antibody/cell line.. Creative Biolabs has updated the generally applied mammalian cell expression with glyco-engineered Chinese hamster ovary (CHO) cells and glyco-engineered human embryonic kidney 293 (HEK293) cells for glycoprotein production, which can guarantee the natural folding and sugar chain composition of glycoprotein, and ...
Glycoengineering of the methylotrophic yeast Hansenula polymorpha for the production of glycoproteins with trimannosyl core N-glycan by blocking core oligosaccharide assembly / Doo-Byoung Oh; J S Park; Moo Woong Kim; Seon Ah Cheon; Eun Jung Kim; Hye Yun Moon; Oh Suk Kwon; Sang Ki Rhee; Hyun Ah Kang , 2008 ...
In Saccharomyces cerevisiae the activity for the lactate-proton symporter is dependent on JEN1 gene expression. Pichia pastoris was transformed with an integrative plasmid containing the JEN1 gene. After 24 h of methanol induction, Northern and Western blotting analyses indicated the expression of JEN1 in the transformants. Lactate permease activity was obtained in P. pastoris cells with a Vmax of 2.1 nmol·s−1·mg of dry weight−1. Reconstitution of the lactate permease activity was achieved by fusing plasma membranes of P. pastoris methanol-induced cells with Escherichia coli liposomes containing cytochrome c oxidase, as proton-motive force. These assays in reconstituted heterologous P. pastoris membrane vesicles demonstrate that S. cerevisiae Jen1p is a functional lactate transporter. Moreover, a S. cerevisiae strain deleted in the JEN1 gene was transformed with a centromeric plasmid containing JEN1 under the control of the glyceraldehyde-3-phosphate dehydrogenase constitutive promotor. ...
Here we describe a new technical solution for optimization of Pichia pastoris shake flask cultures with the example of production of stable human type II collagen. Production of recombinant proteins in P. pastoris is usually performed by controlling gene expression with the strong AOX1 promoter, which is induced by addition of methanol. Optimization of processes using the AOX1 promoter in P. pastoris is generally done in bioreactors by fed-batch fermentation with a controlled continuous addition of methanol for avoiding methanol toxification and carbon/energy starvation. The development of feeding protocols and the study of AOX1-controlled recombinant protein production have been largely made in shake flasks, although shake flasks have very limited possibilities for measurement and control. By applying on-line pO2 monitoring we demonstrate that the widely used pulse feeding of methanol results in long phases of methanol exhaustion and consequently low expression of AOX1 controlled genes. Furthermore, we
Non-glycosylated, recombinant human granulocyte colony-stimulating factor (rhG-CSF), produced by Escherichia coli(filgrastim, leukostim) is widely used to treat a number of serious human diseases...
Methylotrophic yeast species (e.g. Hansenula polymorpha, Pichia pastoris) can grow on methanol as sole source of carbon and energy. These organisms are important cell factories for the production of recombinant proteins, but are also used in fundamental research as model organisms to study peroxisome biology. During exponential growth on glucose, cells of H. polymorpha typically contain a single, small peroxisome that is redundant for growth while on methanol multiple, enlarged peroxisomes are present. These organelles are crucial to support growth on methanol, as they contain key enzymes of methanol metabolism. In this study, changes in the transcriptional profiles during adaptation of H. polymorpha cells from glucose- to methanol-containing media were investigated using DNA-microarray analyses. Two hours after the shift of cells from glucose to methanol nearly 20% (1184 genes) of the approximately 6000 annotated H. polymorpha genes were significantly upregulated with at least a two-fold differential
Optimising yeast as a host for recombinant protein production (review) / Nicklas Bonander and Roslyn M. Bill -- Which yeast species shall I choose? : Saccharomyces cerevisiae versus Pichia pastoris (review) / Richard A.J. Darby [and others] -- Preparation of Pichia pastoris expression plasmids / Christel Logez [and others] -- Preparation of Saccharomyces cerevisiae expression plasmids / David Drew and Hyun Kim -- Codon optimisation for heterologous gene expression in yeast / Kristina Hedfalk -- Yeast transformation to generate high-yielding clones / Mohammed Jamshad and Richard A.J. Darby -- Screening for high-yielding Pichia pastoris clones : the production of G protein-coupled receptors as a case study / Shweta Singh [and others] -- Screening for high-yielding Saccharomyces cerevisiae clones: using a green fluorescent protein fusion strategy in the production of membrane proteins / David Drew and Hyun Kim -- The effect of antifoam addition on protein production yields / Sarah J. Routledge and ...
Inulin is linear oligosaccharides [1]. Although inulin can be hydrolyzed to monosaccharides non-enzymatically by acid treatment, inhibitory by-products produced that complicate downstream biological process. Therefore, it is pivotal to establish a cost-effective inulinase (EC 3.2.1.7) production. In this study, we isolated the DNA sequence encoding the mature peptide sequence of the exo-inulinase gene from Kluyveromyces marxianus CBS 6556 and expressed in Pichia pastoris X-33. The purified recombinant enzyme (reINU) gave a specific activity of 13100 U•mg−1, which was about 100-fold higher that reported for the wild type protein isolated from K. marxianus CBS6556 [2, 3]. Ferguson plot analysis showed that secretion expressed reINU was a 169 kDa dimer. Detailed analytical assays showed that the optimal temperature and pH for reINU were 60 oC and pH 4.0, respectively. It was found that hydrolysis activity was about 20% higher in the presence of Mn2+. reINU was stable for over 3 days at room ...
A lipase gene (atl) was cloned from Aspergillustamarii FS132 for the first time. The gene was found to have an open reading frame of 1024 base pairs (bp), and the coding region of the gene contained two introns (51 bp and 52 bp). Multi-alignment analysis of the deduced amino acid sequence indicated high homology between the enzyme and mono- and diacylglycerol lipases from fungi Aspergillus. The recombinant lipase was expressed in Pichia pastoris GS115 cells. The recombinant lipase was found to have a molecular mass of 36.7 kDa, and it exhibited lipase activity of 20 U/mL in culture supernatant when tributyrin was used as the substrate.
VTU Technology to offer unique combination of technologies for the high-level production of proteins with human-like glycosylation using Pichia GlycoSwitch®. Tucson, Arizona and Grambach, Austria, December 03, 2014 - /EPR BIOTECH NEWS/ - VTU Technology and Research Corporation Technologies (RCT) announced today that they have entered into a development and commercialization agreement for VTU to combine both companies´ Pichia pastoris protein production technologies. Pichia GlycoSwitch® - a new expression system for the production of glycoproteins with human-like glycosylation patterns - and VTU´s yield enhancing Pichia pastoris expression platform are now available in one package from VTU. This unique combination of technologies acting synergistically is of great benefit for customers as it leads to a high performance production platform for recombinant glycoproteins with superior product yields and uniform Man5 - or other human-like glycoforms.. Under the terms of the agreement, RCT grants ...
Jadhav, V., Hackl, M., Druz, A., Shridhar, S., Chung, C-Y., Heffner, K.M., Kreil, D.P., Betenbaugh, M., Joseph Shiloach, J., Niall Barron, N., Grillari, J., Borth, N. (2013) CHO microRNA engineering is growing up: Recent successes and future challenges. Biotechn. Advances, 31:8:1501-1513. Jadhav, V., Hackl, M., Klanert, G., Hernandez Bort, J.A., Kunert, R., Borth, N., Grillari, J. (2014) Stable overexpression of miR-17 enhances recombinant protein production of CHO cells. J Biotechn 175, 38-44. Klug, L., Tarazona, P., Gruber, C., Grillitsch, K., Gasser, B., Trötzmüller, M., Köfeler, H., Leitner, E., Feussner, I., Mattanovich, D., Altmann, F., Daum, G., (2013) The lipidome and proteome of microsomes from the methylotrophic yeast Pichia pastoris, Biochimica et Biophysica Acta, 215-226. Klavins, K., Neubauer, S., Al Chalabi, A., Sonntag, D., Haberhauer-Troyer, C., Russmayer, H., Sauer, M., Mattanovich, D., Hann, S., Koellensperger, G. (2013) Interlaboratory comparison for quantitative primary ...
With a variety of physiological and pharmacological functions, menaquinone is an essential prenylated product that can be endogenously converted from phylloquinone (VK1) or menadione (VK3) via the expression of Homo sapiens UBIAD1 (HsUBIAD1). The methylotrophic yeast, Pichia pastoris, is an attractive expression system that has been successfully applied to the efficient expression of heterologous proteins. However, the menaquinone biosynthetic pathway has not been discovered in P. pastoris. Firstly, we constructed a novel synthetic pathway in P. pastoris for the production of menaquinone-4 (MK-4) via heterologous expression of HsUBIAD1. Then, the glyceraldehyde-3-phosphate dehydrogenase constitutive promoter (PGAP) appeared to be mostsuitable for the expression of HsUBIAD1 for various reasons. By optimizing the expression conditions of HsUBIAD1, its yield increased by 4.37 times after incubation at pH 7.0 and 24 °C for 36 h, when compared with that under the initial conditions. We found HsUBIAD1
Pichia pastoris recombinant protein expression platform. Automated purification by affinity chromatography on Ni-NTA and enzymatic activities assays of His6-tag
The PPIC Statewide Survey delivers objective, advocacy-free information on the perceptions, opinions, and public policy preferences of California residents. PPIC invites input, comments, and suggestions from policy and public opinion experts and from its own advisory committee, but survey methods, questions, and content are determined solely by the PPIC survey team. The PPIC Statewide Survey relies on a rigorous survey methodology and is a charter member of the American Association for Public Opinion Research Transparency Initiative. The survey is conducted regularly throughout the year in the key areas of government, the environment, K-12 education, and higher education ...
Çalık P, Bozkurt B, Zerze GH, İnankur B, Bayraktar E, Boy E, Orman MA, Açık E and Özdamar, TH., Effect of co-substrate sorbitol different feeding strategies on human growth hormone production by recombinant Pichia pastoris. J. Chem. Technol. Biotechnol., 88: 1631-1640. , ...
Production of heterologous proteins in yeast becomes more and more important in the biotechnological sector for research use, therapeutics or sustainable energy sources [1, 2]. This development has led to the constant demand for improvement of hosts and processes for recombinant protein production (RPP). Common approaches for improvement of RPP are mostly based on strain engineering [3], process optimization [4] including adaptation of culture conditions [5-8], feed strategies [9], or media optimization [10, 11]. As production processes become more and more specific due to special needs of production hosts [11] and the recombinant product, media need to be developed for a specific process and are often tailor-made [12]. In general, cultivation media should be chemically defined, however, high level protein synthesis, influencing cell physiology, might make supplementation with e.g. amino acids necessary [13]. For bioreactor cultivations of the yeast production host Pichia pastoris (Komagataella ...
ABSTRACT: BACKGROUND: Inducible high-level expression is favoured for recombinant protein production in Pichia pastoris. Therefore, novel regulated promoters are desired, ideally repressing heterologous gene expression during initial growth and enabl
Expanding the application of technical enzymes, e.g., in industry and agriculture, commands the acceleration and cost-reduction of bioprocess development. Microplates and shake flasks are massively employed during screenings and early phases of bioprocess development, although major drawbacks such as low oxygen transfer rates are well documented. In recent years, miniaturization and parallelization of stirred and shaken bioreactor concepts have led to the development of novel microbioreactor concepts. They combine high cultivation throughput with reproducibility and scalability, and represent promising tools for bioprocess development. Parallelized microplate cultivation of the eukaryotic protein production host Pichia pastoris was applied effectively to support miniaturized phenotyping of clonal libraries in batch as well as fed-batch mode. By tailoring a chemically defined growth medium, we show that growth conditions are scalable from microliter to 0.8 L lab-scale bioreactor batch cultivation ...
HARMSEN, M. C., HEERINGA, P., VAN DER GELD, Y. M., HUITEMA, M. G., KLIMP, A., TIRAN, A. and KALLENBERG, C. G. M. (1997), Recombinant proteinase 3 (Wegeners antigen) expressed in Pichia pastoris is functionally active and is recognized by patient sera. Clinical & Experimental Immunology, 110: 257-264. doi: 10.1111/j.1365-2249.1997.tb08325.x ...
Vyepti was the 4th and most recent of the CGRP mAbs to become available. It is made by Lundbeck and was FDA approved for migraine prevention 2/21/20. The antibody is produced in Pichia pastoris yeast cells by recombinant DNA technology. It targets the CGRP ligand, rather than the CGRP receptor. It binds very strongly to the CGRP ligand, interfering with its ability to bind to the CGRP receptor and activate the migraine. It comes in 100 mg and 300 mg doses and is dosed once quarterly (every 3 months) by a quick 30-minute infusion. The 100 mg dose is the recommended starting dose which can be titrated as needed to the higher dose later.. This is the only intravenous (IV) option available. Since it is administered IV, it is 100% bioavailable compared to the bioavailability of the other subcutaneous injections of 50-82%. It also reaches Cmax (maximum concentration) in about 30 minutes compared to 5-7 days of the other subcutaneous injections. Therefore, not surprisingly Vyepti showed treatment ...
Derived from Pichia Pastoris yeast, the patented fermentation technology produces a non-allergenic, high-performance Vegan registered collagen
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The methylotrophic yeast P. pastoris has, during the last decades, increased in popularity as a eukaryotic host for recombinant protein expression [7]. Numerous reports have described successful overexpression of soluble proteins as well as of membrane proteins in this system, but sufficient protein yields have not always been obtained. Several methods have been described that could be used to improve the outcome of expression trials, among which an optimisation of recombinant gene dosage has proven to be one of the most potent ones [21]. When it comes to the aquaporin family of membrane proteins, optimisation of the nucleotide sequence both regarding codon composition and AT content [38] and controlled growth in bioreactors [36] have been reported to improve the yields of heterologous protein. However, in no reports has a systematic examination of the effect of gene dosage on recombinant aquaporin expression in P. pastoris been done and results obtained for other membrane proteins [12, 15, 33] ...
Advanced tumor growth is angiogenesis dependent (27) and endothelial cell organization into vascular networks is a hallmark of the angiogenic process (28). Such organization involves the mobilization of endothelial cells from a quiescent state to form a quasi-mature vasculature that can supply the growing mass with nutrients and oxygen (28). Inhibition of endothelial cell tube formation by endostatin may be due (at least in part) to blockade of the endothelial cells mobilization stimuli, although it is unclear whether this inhibition requires direct interaction with the endothelial cell or other cells that provide the required stimulatory cytokines. Surprisingly, endostatin obtained from EntreMed or Calbiochem had different efficacy profiles in our endothelial cell tube formation assays. This observation was unexpected given that EntreMed produces human endostatin in a P. pastoris expression system and supplies it to Calbiochem for packaging and sale. Follow-up experiments were conducted to ...
Recombinant protein production lies at the core of the biotechnology revolution. Recombinant proteins such as humanized chimeric antibodies, human growth hormone, human insulin and a variety of industrial enzymes can been expressed in the yeast Pichia pastoris, a premier organism for eucaryotic protein expression. Pichia pastoris grows rapidly on inexpensive substrates to very high cell densities, and can produce biologically active foreign proteins of higher eukaryotic origin since it performs important posttranslational protein modifications, including proteolytic processing, disulfide bridge formation and glycosylation.. Regenerative medicine is aimed at using biological solutions to restore function to damaged or diseased biological systems. Work in this field could potentially yield viable tissues or organs for replacement, as well as therapies for a number of diseases. Stem cell therapies and gene therapy are among the most promising research avenues in regenerative medicine. Stem cells ...
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Fibroblast Growth Factor 21 (FGF21) is a novel target with potential anti diabetic properties that are useful for treatment of hyperglycemia, insulin resistance, hyperlipidemia and metabolic disease. Producing recombinant FGF21 by E. coli without using fusion proteins is time consuming and will produce low quantity products. In this study, to establish and test the efficiency of other expression methods, the complete fgf21 gene was constructed by overlapping PCR. The recombinant fgf21 genes were expressed successfully in E. coli (TB1) and in yeast (Pichia pastoris) under the control of maltose binding promoter and alcohol oxidase I promoter. The degree of success in terms of yield and functionality of the produced recombinant proteins in vivo were compared by using animal models. The result demonstrated that both expression systems can promote more soluble FGF21 levels, with less purification steps while preserving the bioactivity of the protein in vivo. The FGF21 produced in P. pastoris ...
in Journal of industrial microbiology & biotechnology (2016), 43(4), 517-23. High Pichia pastoris biomass density could be obtained using high co-feeding rate of methanol and sorbitol in a fed-batch or continuous culture, while further higher feeding rate finally leads to oxygen ... [more ▼]. High Pichia pastoris biomass density could be obtained using high co-feeding rate of methanol and sorbitol in a fed-batch or continuous culture, while further higher feeding rate finally leads to oxygen limitation in bioreactor. In the literature, there is lack of report about AOX1 promoter regulation with regard to dissolved oxygen level (DO). Therefore, in this work, chemostat cultures were performed to investigate the cell growth, metabolism and regulation of the AOX1 promoter (pAOX1) regarding co-feeding rate of optimized methanol/sorbitol mixture (methanol fraction 0.60 C-mol/C-mol) using a P. pastoris Mut+/pAOX1-lacZ strain. The oxygen transfer rates (OTR) in bioreactor were kept in the range of ...
Saccharomyces cerevisiae and Pichia pastoris are two of the most relevant microbial eukaryotic platforms for the production of recombinant proteins. Their known genome sequences enabled several transcriptomic profiling studies under many different environmental conditions, thus mimicking not only perturbations and adaptations which occur in their natural surroundings, but also in industrial processes. Notably, the majority of such transcriptome analyses were performed using non-engineered strains. In this comparative study, the gene expression profiles of S. cerevisiae and P. pastoris, a Crabtree positive and Crabtree negative yeast, respectively, were analyzed for three different oxygenation conditions (normoxic, oxygen-limited and hypoxic) under recombinant protein producing conditions in chemostat cultivations. The major differences in the transcriptomes of S. cerevisiae and P. pastoris were observed between hypoxic and normoxic conditions, where the availability of oxygen strongly affected
I have a recombinant protein produced in the yeast Pichia pastoris that is partially glycosylated with O-linked sugar (it must be O-linked - there are no N-linked sites). I wish to remove the glycosylated protein from the non-glycosylated. I have tried using ConA-Sepharose (Pharmacia) and Glycine-Max (Sigma). Neither of these has appeared to bind. The ConA comes with instructions that have been followed. No instructions could be found for Gycine-MAx and Sigma were not forthcoming. Has anyone has experience with trying to purify O-linked sugars, with either of the above lectins, or using any other reagents, I would very much like to hear from you. Please reply to me by e-mail as I do not regularly read this newsgroup. Tim -- Tim Dudgeon British Biotech Phone: 0865 748747 FAX: 0865 717598 email: dudgeon at britbio.co.uk ...
Gentaur molecular products has all kinds of products like :search , Ray Biotech \ Recombinant Human Granulocyte Macrophage-Colony Stimulating Factor, Pichia \ 228-10556-2 for more molecular products just contact us
151732DNAPichia pastorisgene(1)..(732) 1atgagtttgg ttgcacttca tcaattcgac tacatctttg cgatcgccat gatgtttgca 60 ttcttggatg cattcaacat cggcgcaaat gatgtagcaa actccttttc ttcatcagtc 120 tcctcgagat gtctgaaata ctggcaagcg atgatcttag ctgccatcat ggaatttttg 180 ggtgccgtct tagctggtgc ccgtgtcact gacactatta gaaagagaat tatcaacgtt 240 cacgctttcg acgacgaccc agccatgctg atgttgacca tggccactgc cttggtcggt 300 tcttctgtct ggttgagtat tgccacttat gtcagagctc ctgtctccac cactcattcc 360 attgttggtg gtgtcattgg tgctggtatc gctgctaaag gtgctggtga gatccactgg 420 ggatggagtg gatttgccaa gattgttgct tcttggttca tcgctccatg tgttgctggt 480 ggattcgctt ccgtcctttt cctcttctgt aagttctcaa ttttggagag aaagcatgat 540 gccagaaatg cccttctgtt tgctccatgt atcgtcttct tgacttttgc tgtcctgaca 600 atgttgatcg tctggaaggg agctccaaac ttgaacttgg acgacctttc tactggtgct 660 accgtcggnt ctatcttcgg tgttgctggt gttgctacaa tcatttacat cacattcttt 720 ttccattctt ga 732 225DNAArtificial Sequenceprimer 2gaatacacac aagcatctat ttgag 25 326DNAArtificial Sequenceprimer 3ccactccatc ...
1B3E: X-ray crystallography and mass spectroscopy reveal that the N-lobe of human transferrin expressed in Pichia pastoris is folded correctly but is glycosylated on serine-32.
The methylotrophic yeast, Pichia pastoris, has been genetically engineered to produce many heterologous proteins for industrial and research purposes. In order to secrete proteins for easier purification from the extracellular medium, the coding sequence of recombinant proteins is initially fused to the Saccharomyces cerevisiae α-mating factor secretion signal leader. Extensive site-directed mutagenesis of the prepro-region of the α-mating factor secretion signal sequence was performed in order to determine the effects of various deletions and substitutions on expression. Though some mutations clearly dampened protein expression, deletion of amino acids 57-70, corresponding to the predicted 3rd alpha helix of α-mating factor secretion signal, increased secretion of reporter proteins horseradish peroxidase and lipase at least 50% in small-scale cultures. These findings raise the possibility that the secretory efficiency of the leader can be further enhanced in the future.
A major bottleneck in structural, biochemical and biophysical studies of proteins is the need for large amounts of pure homogenous material, which is generally obtained by recombinant overexpression. Here we introduce a vector collection, the pCri System, for cytoplasmic and periplasmic/extracellular expression of heterologous proteins that allows the simultaneous assessment of prokaryotic and eukaryotic host cells (Escherichia coli, Bacillus subtilis, and Pichia pastoris). By using a single polymerase chain reaction product, genes of interest can be directionally cloned in all vectors within four different rare restriction sites at the 5′end and multiple cloning sites at the 3′end. In this way, a number of different fusion tags but also signal peptides can be incorporated at the N-and C-terminus of proteins, facilitating their expression, solubility and subsequent detection and purification. Fusion tags can be efficiently removed by treatment with site-specific peptidases, such as tobacco ...
P. pastoris and E. coli are commonly used hosts for recombinant product expression because of their ability to produce large quantities of recombinant protein quickly and economically. P. pastoris can express proteins with correct primary, secondary and tertiary structure and post-translational modifications such as glycolysation, proteolytic processing and disulfide bond formation. E. coli are readily transformed and can express large quantities of proteins quickly with correct primary structure in the soluble or insoluble forms. These attributes ensure that each host will remain central in recombinant protein expression of the future ...
Interleukin-8 Human Recombinant produced in Yeast is a single, glycosylated polypeptide chain containing 77 amino acids and having a molecular mass of 8904 Dalton.
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IBI recombinant proteins are expressed in Pichia pastoris (yeast) system which provides many advantages over E. coli systems. Yeast systems promote proper prote
ABSTRACT The PaEXG2 gene, encoding an exo-beta-1,3-glucanase, was isolated from the biocontrol agent Pichia anomala strain K… Expand ...
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The methylotrophic yeast Pichia pastoris can be used to express recombinant genes at high levels under the control of the methanol-inducible alcohol oxidase 1 (AOX1) promoter. Accurate regulation of the methanol concentration in P. pastoris cultures is necessary to maintain induction, while preventing accumulation of methanol to cytotoxic levels. We developed an inexpensive methanol sensor that uses a gas-permeable silicone rubber tube immersed in the culture medium and an organic solvent vapor detector. The sensor was used to monitor methanol concentration continuously throughout a fed-batch shake-flask culture of a P. pastoris clone producing the N-lobe of human transferrin. The sensor calibration was stable for the duration of the culture and the output signal accurately reflected the methanol concentration determined off-line by HPLC. A closed-loop control system utilizing this sensor was developed and used to maintain a 0.3% (v/v) methanol concentration in the culture. Use of this system ...
Yeast Pichia stipitis CBS 5776 was developed through adaptation fermentation step by step in steam exploded corn stover prehydrolyzate because high concentration of weak acids and other inhibitors present in the prehydrolyzate could degrade the fermentability. However, the adaptability of Pichia stipitis CBS 5776 in the prehydrolyzate was so limited that steam strip-ping and overliming were applied to remove these inhibitors from it. Corn stover was steam exploded; the filtrate of steam exploded corn stover was hydrolyzed with dilute sulfuric acid, and then the acid hydrolyzate was detoxified and fermented by Pichia stipitis CBS 5776. Steam stripping could remove volatile com-pounds from the acid hydrolyzate and the fil-trate. At a steam stripping time of 120min, 81% acetic acid and 59% formic acid were removed from the acid hydrolyzate, 77% acetic acid and 45% formic acid were removed from the filtrate, while furfural was stripped off completely from the acid hydrolyzate and the filtrate. Overliming
TY - JOUR. T1 - Codon optimization of Candida rugosa lip1 gene for improving expression in Pichia pastoris and biochemical characterization of the purified recombinant LIP1 lipase. AU - Chang, Shu Wei. AU - Lee, Guan Chiun. AU - Shaw, Jei Fu. N1 - Copyright: Copyright 2011 Elsevier B.V., All rights reserved.. PY - 2006/2/8. Y1 - 2006/2/8. N2 - An important industrial enzyme, Candida rugosa lipase (CRL) possesses several different isoforms encoded by the lip gene family (lip1-lip7), in which the recombinant LIP1 is the major form of the CRL multigene family. Previously, 19 of the nonuniversal serine codons (CTG) of the lip1 gene hav been successfully converted into universal serine codons (TCT) by overlap extension PCR-based multiple-site-directed mutagenesis to express an active recombinant LIP1 in the yeast Pichia pastoris. To improve the expression efficiency of recombinant LIP1 in P. pastoris, a regional synthetic gene fragment of lip1 near the 5′ end of a transcript has been constructed to ...
The gene encoding lipase B from Candida antarctica (CalB) was expressed in Pichia pastoris after it was synthesized by the recursive PCR and cloned into the Pichia expression plasmid, pPICZαA. The CalB was successfully secreted in the recombinant P. pastoris strain X-33 with an apparent molecular weight of 34 kDa. For 140 h flask culture, the dry cell weight and the extracellular lipase activity reached at 5.4 g/l and 57.9 U/l toward p-nitrophenyl palmitate, respectively. When we performed the fed-batch fermentation using a methanol feeding strategy for 110 h, the dry cell weight and the extracellular lipase activity were increased to 135.7 g/l and 11,900 U/l; the CalB protein concentration was 1.18 g/l of culture supernatant. The characteristics of CalB recovered from the P. pastoris culture were compared with the commercial form of CalB produced in Aspergillus oryzae. The kinetic constants and specific activity, the effects of activity and stability on temperature and pH, the glycosylation extent,
Medium-chain volatile flavour esters are important molecules since they have extensive applications in food, fragrance, cosmetic, paint and coating industries, which determine different characteristics of aroma or taste in commercial products. Biosynthesis of these compounds by alcoholysis is catalyzed by acyl-CoA:ethanol O-acyltransferases Eht1 or Eeb1 in Saccharomyces cerevisiae. In this study, these two yeast enzymes were selected to explore their preparations as the form of whole cell biocatalysts for the production of volatile flavour esters. Here, the novel whole cell biocatalysts Pichia pastoris yeasts with functional extracellular expression of Eht1 or Eeb1 were constructed. Flavour production was established through an integrated process with coupled enzyme formation and ester biosynthesis in the recombinant yeasts in one pot, leading to the formation of volatile C6-C14 methyl and ethyl esters from wort medium. Interestingly, there is no significant difference between P. pastoris-EHT1 ...
The endo-β-1,4-glucanase gene celE from the anaerobic fungus Orpinomyces PC-2 was placed under the control of an alcohol oxidase promoter (AOX1) in the plasmid pPIC9K, and integrated into the genome of a methylotrophic yeast P. pastoris GS115 by electroporation. The strain with highest endo-β-1,4-glucanase activity was selected and designed as P. pastoris egE, and cultivated in shaking flasks. The culture supernatant was assayed by SDS-polyacrylamide gel electrophoresis and showed a single band at about 52 kDa. Furthermore, the recombinant P. pastoris egE was proved to possess the ability to utilize sodium carboxymethyl cellulose as a carbon source. The recombinant endoglucanase produced by P. pastoris showed maximum activity at pH 6.0 and temperature 45 °C, indicating it was a mesophilic neutral endo-β-1,4-glucanase, suitable for denim biofinishing/washing. Further research was carried out in suitable fermentation medium in shaking flasks. The most favorable methanol addition concentration was
The intracellular metabolic fluxes through the central carbon pathways in the bioprocess for recombinant human erythropoietin (rHuEPO) production by Pichia pastoris (Mut(+)) were calculated. to investigate the metabolic effects of dual carbon sources (methanol/sorbitol) and the methanol feed rate, and to obtain a deeper understanding the regulatory circuitry of P. pastoris, using the established stoichiometry-based model containing 102 metabolites and 141 reaction fluxes. Four fed-batch operations with (MS-) and without (M-) sorbitol were performed three different constant specific growth rates (h(-1)), and denoted as M-0.03, MS-0.02, MS-0.03, and MS-0.04. Considering the methanol consumption pathway, the M-0.03 and MS-0.02 conditions produced similar effects and had >85% of formaldehyde flux towards the assimilatory pathway. In contrast, the use of the dual carbon Source condition generated la shift in metabolism towards the dissimilatory pathway that corre- sponded to the shift in dilution ...
Recently, we engineered Pichia pastoris Muts strains to produce several beta-propeller phytases, one from Bacillus subtilis and the others designed by a structure-guided consensus approach. Furthermore, we demonstrated the ability of P. pastoris to produce and secrete these phytases in an active form in shake-flask cultures. In the present work, we used a design of experiments strategy (Simplex optimization method) to optimize five environmental factors that define the culture conditions in the induction step to increase beta-propeller phytase production in P. pastoris bioreactor cultures. With the optimization process, up to 347,682 U (82,814 U/L or 6.4 g/L culture medium) of phytase at 68 h of induction was achieved. In addition, the impact of the optimization process on the physiological response of the host was evaluated. The results indicate that the increase in extracellular phytase production through the optimization process was correlated with an increase in metabolic activity of P. ...
Deschampsia antarctica shows tolerance to extreme environmental factors such as low temperature, high light intensity and an increasing UV radiation as result of the Antarctic ozone layer thinning. It is very likely that the survival of this species is due to the expression of genes that enable it to tolerate high levels of oxidative stress. On that account, we planned to clone the D. antarctica Cu/ZnSOD gene into Pichia pastoris and to characterize the heterologous protein. The Copper/Zinc superoxide dismutase (Cu/ZnSOD) gene, SOD gene, was isolated from a D. antarctica by cDNA library screening. This SOD gene was cloned in the expression vector pGAPZαA and successfully integrated into the genome of the yeast P. pastoris SMD1168H. A constitutive expression system for the expression of the recombinant SOD protein was used. The recombinant protein was secreted into the YPD culture medium as a glycosylated protein with a 32 mg/l expression yield. The purified recombinant protein possesses a specific
Evaluation of fermentation kinetics of xylose to ethanol fermentation in the presence of acetic acid by Pichia stipitis: Modeling and experimental data comparison
Different cultivation strategies have been compared for the production of Rhizopus oryzae lipase (ROL) from Pichia pastoris. Several drawbacks have been found using a methanol non-limited fed-batch. On the one hand, oxygen limitation appeared at early cell dry weights and, on the other hand, high cell death was observed. A temperature limited fed-batch has been proposed to solve both problems. However, in our case study a methanol non-limited fed-batch results in better productivities. Finally, a lower salt medium were used to overcome cell death problems and a temperature limited fed-batch was applied thereafter to solve oxygen transfer limitations. This combined strategy has resulted in lower productivities when compared to a methanol non-limited fed-batch. However the culture could be longer prolonged and a 1.3-fold purer final product was obtained mainly due to cell death reduction.. ...
TY - CHAP. T1 - Heterologous expression of isotypically labelled Trichoderma reesei tyrosinase 2 in Pichia pastoris. AU - Westerholm-Parvinen, Ann. AU - Mattinen, Maija. AU - Selinheimo, Emilia. AU - Markku, Saloheimo. PY - 2007. Y1 - 2007. N2 - Tyrosinase (EC 1.14.18.1) is a copper-containing oxidase that is widely distributed in mammals, invertebrates, plants and microorganisms. In mammals the enzyme is essential for the formation of melanin pigments, whereas tyrosinases in fruit and vegetables are related to the browning reaction that occurs upon bruising and long term storage. Tyrosinase is of great interest for many applications in the field of medicine, biotechnology and food engineering. It is a promising target enzyme for prodrug activations in melanomas and in biotechnological applications including crosslinking of protein matrices. It is of great importance to find ligands and inhibitors for tyrosinase. Structural studies and screening for ligands and inhibitors can be carried out ...
Search and download thousands of Swedish university dissertations (essays). Full text. Free. Dissertation: Bench-Scale Production of Heterologous Proteins from Extremophiles- Escherichia coli and Pichia pastoris based expression systems.
Article Biochemical Conversion of Acid-Pretreated Water Hyacinth (Eichhornia Crassipes) to Alcohol Using Pichia Stipitis NCIM3497. Cleaning and removal of water hyacinth from lakes and various historical places government spends lacks of rupees per y...
The dimorphic yeast Candida rugosa has an unusual codon usage that hampers the functional expression of genes derived from this yeast in a conventional heterologous host. Commercial samples of C. rugosa lipase (CRL) are widely used in industry, but contain several different isoforms encoded by the lip gene family, among which the isoform encoded by the gene lip1 is the most prominent. In a first laborious attempt, the lip1 gene was systematically modified by site-directed mutagenesis to gain functional expression in Saccharomyces cerevisiae. As alternative approach, the gene (1647 bp) was completely synthesized with an optimized nucleotide sequence in terms of heterologous expression in yeast and simplified genetic manipulation. The synthetic gene was functionally expressed in both hosts S. cerevisiae and Pichia pastoris, and the effect of heterologous leader sequences on expression and secretion was investigated. In particular, using P. pastoris cells, the synthetic gene was functionally ...
Scheffersomyces stipitis ATCC ® 58785™ Designation: CBS 6054 [CCRC 21777, IFO 10063, NRRL Y-11545] Application: Bioethanol production
Scheffersomyces stipitis ATCC ® 58785D-2™ Designation: genomic DNA from CBS 6054 (ATCC 58785) Application: Bioethanol production
1311003DNAPichia pastoris 1tgccaggatc tgaccactac aagaagcgta attaagtatt tattatgtag cttgaggaat 60tgatttctaa aggccaaaga tgatgtagag caatagtaga gtgattagca agtttctgaa 120catgttttgt ggtcgggtat taggcgccgt acctctgtat tttttttttt tggcagatgg 180gatgaccact gtgcttacat aattttcgat gggccctttt ggaaacgacc actccttccg 240atcacgtaat actaggcgag tatcactacg cgtcggtatc cgcattttag ccgctttaac 300gacaaggtca gggtaaatac tgtgttgcat gggcgatgcc gtaggatctg gccaacttgg 360aacagagcag cccaccgcgt aggcagcgct tgggaccagt ttgttagcca atcagttccc 420ccgtctgagt ccctacacgc tcaggcttgc atggatatgc atgaaaaatg tcaactcagt 480cgcacacagg gatgctgttg tcgtggtgag aaaaagctaa aatagcaaac ccatagcaat 540cgagagactg tgattggaag caaaagaacc aggcattgac taggaaagac tgacaagaaa 600attagacggt gccagtggta tttcattgaa ttgcactgaa cacgcctccc atttccttcc 660ctcaatccat tcaaagggct aattaaacct tatctcggcc tacattagtt gtcatggaaa 720cgtctcactt tctctctgtg tcccacagta gcgagtaggc ggagagacca aaaatggtct 780ctcggcccta atcaaaacat cctttgtcgc gttgttttca gatcgattcg caatctgaga 840gaaaaacaat ...
Did you know that Louis Pasteur discovered microorganisms and developed his technique for killing them, pasteurization, while studying beer? He was also a master brewer and wrote a book on the subject. Beer and life science have a long history, indeed even enzymes were discovered by studying yeast fermentation.. In San Diego, we are lucky to be a hub for both biotech and beer, and weve found a very interesting intersection of the two in White Labs, a local company creating high quality yeast strains for breweries. The company was started by Chris White, who got a Ph.D. in Biochemistry at UCSD studying the yeast Pichia pastoris. Chris was also an avid home beer brewer and began to use his scientific expertise on brewers yeast. We will hear from him about how he used this experience to found White Labs, a creative entrepreneurial move that has led to biotech helping to develop San Diegos thriving beer industry.. Of course, weve been holding SDBN events at Green Flash Brewery for almost a year, ...
سال انتشار: ۱۳۸۶ محل انتشار: پنجمین کنگره بین المللی مهندسی شیمی تعداد صفحات: ۸ نویسنده(ها): Bahrami - Biotechnology Group, Chemical Engineering Department,
Background: Over the past century, the areas of genomics, proteomics and lipids have captured the attention of investigators worldwide. Carbohydrates, have recently received increased attention through the expanding field of glycobiology; probably because they are very complex and not encoded in the genome. Objectives: The purpose of this study was to express and purify recombinant human galectin 3via the Pichiapastoris expression system. Materials and Methods:cDNA of human galectin 3 gene was amplified with specific primers and cloned into a pcDNA3.1 vector with His-tag for easier purification using Ni2andchromatography. Furthermore, galectin 3was purified to homogeneity and confirmed using SDS-PAGE and western blotting. Results:The protein band corresponding to 29 kDa was excised from the gels, digested with trypsin and processed for mass spectrometric analysis by Matrix Assisted Laser Desorption/Ionization- Time of Flight Mass Spectroscopy (MALDI-TOF MS), using a Reflex III instrument.
Many filamentous fungi and yeast species have been used for heterologous protein. This chapter covers the most commonly used fungal expression systems (Saccharomyces cerevisiae, Pichia pastoris, and Aspergillus species), with a focus on vector systems, promoter and leader sequences, posttranslational modifications, and fermentation scalability. Similar to other heterologous protein expression systems, proteolytic hydrolysis often causes instability of foreign proteins in Pichia. In comparison with S. cerevisiae and P. pastoris, filamentous fungi are less studied for heterologous protein production with respect to their transcription and translation control, mRNA stability, glycosylation, and regulation of protein secretion. There are several web-accessible organizations such as the American Type Culture Collection (ATCC), the (Fungal Genetics Stock Center (FGSC), the Agricultural Research Service Culture Collection (NRRL), and the Belgium Culture Collection (BCCP) from which the necessary tools for
ID PISTI1_1_PE967 STANDARD; PRT; 362 AA. AC PISTI1_1_PE967; A3GHJ2; DT 00-JAN-0000 (Rel. 1, Created) DT 00-JAN-0000 (Rel. 2, Last sequence update) DT 00-JAN-0000 (Rel. 3, Last annotation update) DE Flags: Fragments; DE SubName: Full=Predicted protein;Flags: Fragment; (PISTI1_1.PE967). GN ORFNames=PICST_53517; OS SCHEFFERSOMYCES STIPITIS CBS 6054. OC Eukaryota; Fungi; Dikarya; Ascomycota; Saccharomycotina; Saccharomycetes; OC Saccharomycetales; Debaryomycetaceae; Scheffersomyces. OX NCBI_TaxID=322104; RN [0] RP -.; RG -.; RL -.; CC -!- SEQ. DATA ORIGIN: Translated from the HOGENOM CDS PISTI1_1.PE967. CC Pichia stipitis (strain CBS 6054 / NRRL Y-11545 / IFO 10063 / ATCC 5878 CC chromosome 1, complete sequence. CC -!- ANNOTATIONS ORIGIN:A3GHJ2_PICST CC -!- SIMILARITY: Contains 1 protein kinase domain. CC -!- GENE_FAMILY: HOG000233016 [ FAMILY / ALN / TREE ] DR UniProtKB/Swiss-Prot; A3GHJ2; -. DR EMBL; AAVQ01000002; EAZ63064.2; -; Genomic_DNA. DR RefSeq; XP_001387087.2; XM_001387050.1. DR ...
Among more than 20 yeast strains isolated from the traditional starter murcha in Nepal, we characterized a yeast that might be involved in saccharification. This strain, identified as ,I,Pichia burtonii,/I,, produced an extracellular amylolytic enzyme when cultured in the presence of starch in the medium. Since no amylase secreted by ,I,P. burtonii,/I, has yet been reported, we purified the enzyme and determined its N-terminal amino acid sequence. Together with the results of a hydrolyzing activity assay toward various substrates, it was found to be an α-amylase. The purified enzyme, named ,I,Pichia burtonii,/I, α-amylase (PBA), was a glycoprotein with an apparent molecular mass of 51 kDa. Enzyme activity was optimal at pH 5.0 at 40 °C. The enzyme retained 80% of its original activity after incubation under the optimal pH condition at 50 °C for 30 min. The activity was inhibited by metal ions such as Cd,SUP,2+,/SUP,, Cu,SUP,2+,/SUP,, Hg,SUP,2+,/SUP,, Al,SUP,3+,/SUP,, and ...
Blog on HBsAg recombinant protein product: The HBsAg n/a (Catalog #MBS434117) is a Recombinant Protein produced from Pichia Pastoris and is intended...
We describe the introduction of the yeasts Saccharomyces cerevisiae and Pichia pastoris as eukaryotic hosts for the routine production of recombinant proteins for a structural genomics initiative. We
Instructions will be provided on presentation format options for your talk or poster once selected for the program. We will try to be as flexible as possible. More than one submission per person is welcome. There is no limit to the number of posters; most any scientific presentation will be accepted. ...
Endostatin Human Recombinant produced in Pichia Pastoris is a single, glycosylated, polypeptide having a total molecular mass of 20,000 Dalton C-terminal.
Abstract: Pancreatic phospholipase A2 (phospholipase A2 group 1B, G1B) belongs to the superfamily of secreted phospholipase A2 (PLA2) enzymes. G1B has been proposed to be a potential target for diseases such as hypertension, obesity, and diabetes. Human pancreatic prophospholipase A2 (pro-hG1B) is activated by cleavage of the first seven-residue propeptide (phospholipase A2 propeptide, PROP). However, questions still remain on the mode of action for pro-hG1B. In this work, we expressed pro-hG1B in Pichia pastoris and determined the crystal structure at 1.55-Å resolution. The x-ray structure demonstrates that pro-hG1B forms a trimer. In addition, PROP occupies the catalytic cavity and can be self-cleaved at 37 °C. A new membrane-bound surface and activation mechanism are proposed based on the trimeric model of pro-hG1B. We also propose a new autoproteolytic mechanism for pro-hG1B by the reaction triad Asp49-Arg0-Ser(-2) that is similar to the serine protease catalytic triad.. ...
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Click Generate. We now have two ranges, both with 3 models (no longer 4) available from 3. A Desktop search for windows server 2008 r2 with Plesk Panel control winvows brings flexibility and control to a new level of cost effective powerful website automation. CISSA is a non-profit, non-governmental organization of scientists and citizens combining rigorous scientific analysis, innovative policy development and effective citizen advocacy to build a cleaner, safer and healthier world. I may install two more software to handle email accounts if I feel dekstop in future. Do you wear glasses. We offer various types of Operating System, from Windows to Linux. In addition, the yeast PLD, Spo14p, is required to assemble sporulation-related prospore membranes under starvation conditions 62 and is involved in AP formation-dependent unconventional secretion of Pichia pastoris Acb1 63 Collectively, these results support the hypothesis that Asp connection string to sql server 2005 express is involved in ...
Alcohol Oxidase, 10 mg. Alcohol Oxidase (AOX) is a homooctameric flavoprotein consisting of eight identical subunits of ~74 kD, each containing a flavin adenine dinucleotide molecule (FAD) as a prosthetic group (van der Klei et al.
Meg Whitman leads Jerry Brown 44% to 39% among likely voters in the race for governor and Brown is favored over Poizner, 46% to 31 ...
Since this Asana is good for leading a life of chastity and continence, persons who are desirous of leading such a life may have suggest to themselve
Steudel A., Nitzsche C.: The nad-dependent alcohol dehydrogenase of methylotrophic yeasts - an electrophoretic study (i). Acta Biotechnol. 1991, 11, 57. ,https://doi.org/10.1002/abio.370110117 ...
Xylaria polymorpha, Dead Mans Fingers fungus: identification pictures (images), habitat, edible or poisonous; taxonomy, etymology, synonyms, similar species
Basel, Switzerland and Melbourne, Australia, 18 Dec 2014 - Lonza and AdAlta announced today that they have entered into a strain development agreement for AdAltas alternative scaffold proteins, called i-bodies. Lonza will conduct strain development studies with the XS™ Pichia systems for several i-body molecules that AdAlta successfully expressed at research levels in their Melbourne-based laboratory. Strain development and manufacturing will be conducted by Lonza at its microbial facility in Visp, Switzerland to establish a commercially viable manufacturing process.. The XS™ Research Evaluation Agreement (REA) provided AdAlta scientists with access to Lonzas proprietary microbial expression systems, protocols and technical expertise. The licensable XS™ Technology Platform consists of multiple hosts and promoter systems that have been optimized for soluble expression driving simpler recovery and downstream processing, which saves development time and costs. After evaluating several ...
Anti-aging / Anti-wrinkle -Biovesicle encapsulated bFGF -INCI Name: Phosphate Buffered Saline (and) Pichia r-sh-Polypeptide-1 Ferment Extract Filtrate -Increasing
Su, Lin-Hui; Ou, Jonathan T.; Leu, Hsieh-Shong; Chiang, Ping-Cherng; Chiu, Yueh-Pi; Chia, Ju-Hsin; Kuo, An-Jing; Chiu, Cheng- ...
Some specialized yeasts, e.g., Candida tropicale, Pichia sp., Rhodotorula sp., can use alkanes as a source of carbon or energy ...
A yeast commonly used for protein production is Pichia pastoris. Examples of yeast expression vector in Pichia are the pPIC ... "Pichia pastoris Expression System" (PDF). Invitrogen. "K. lactis Protein Expression Kit" (PDF). New England BioLabs Inc. Fields ... Cregg JM, Cereghino JL, Shi J, Higgins DR (2000). "Recombinant protein expression in Pichia pastoris". Molecular Biotechnology ...
Expression systems using either S. cerevisiae or Pichia pastoris allow stable and lasting production of proteins that are ... Cregg JM, Cereghino JL, Shi J, Higgins DR (September 2000). "Recombinant protein expression in Pichia pastoris". Molecular ...
Menotti Del Pichia Juliana Carneiro da Cunha...foreign dancer (allusion to Isadora Duncan) Dina Sfat...Branca Clara (allusion ...
In the yeast strain Pichia pastoris, lysyl oxidase constitutes a homodimeric structure. Each monomer consists of an active site ... "The crystal structure of Pichia pastoris lysyl oxidase". Biochemistry. 42 (51): 15148-57. doi:10.1021/bi035338v. PMID 14690425 ...
He leads a team of scientists engaged in the studies of Pichia pastoris, a methylotrophic yeast species, with regard to its ... "Transcriptional interference in the methylotrophic yeast, Pichia pastoris". IISER, Thiruvananthapuram. 2012. "BHD-2011" (PDF). ... Pichia pastoris at Indian Institute of Science Education and Research, Thiruvananthapuram in October 2012. He was the co- ... and pyruvate carboxylase-deficient conditions in Pichia pastoris". Microbiology. 157 (12): 3361-3369. doi:10.1099/mic.0.053488- ...
"GRAS Notification for Soybean Leghemoglobin Protein Derived from Pichia pastoris" (PDF). Archived (PDF) from the original on ...
Murasugi A, Kido I, Kumai H, Asami Y (2003). "Efficient production of recombinant human pleio-trophin in yeast, Pichia pastoris ... Pichia pastoris". Protein Expr. Purif. 27 (2): 244-52. doi:10.1016/S1046-5928(02)00587-9. PMID 12597883. ...
Murasugi A, Kido I, Kumai H, Asami Y (2003). "Efficient production of recombinant human pleio-trophin in yeast, Pichia pastoris ...
Pichia pastoris does have stacked Golgi, while Saccharomyces cerevisiae does not. In plants, the individual stacks of the Golgi ...
Murphy KP, Gagne P, Pazmany C, Moody MD (March 1998). "Expression of human interleukin-17 in Pichia pastoris: purification and ...
"Tomographic Evidence for Continuous Turnover of Golgi Cisternae in Pichia pastoris". Molecular Biology of the Cell. 14 (6): ... "Tomographic evidence for continuous turnover of Golgi cisternae in Pichia pastoris". Molecular Biology of the Cell. 14 (6): ...
Murasugi A, Asami Y, Mera-Kikuchi Y (2001). "Production of recombinant human bile-salt-stimulated lipase in Pichia pastoris". ...
Feng Y, Cui LB, Liu CX, Ma QJ (2001). "[Inhibition effect in vitro of purified endostatin expressed in Pichia pastoris]". Sheng ...
Pichia cactophila (1978) from decaying cacti and Drosophila flies feeding on them. Pichia heedii (1978) from the soft rot of ... Pichia opuntiae (1979) from the cladodes of Opuntia inermis in Australia and from the decaying parts of Cereoid cacti in North ... Starmer, W. T.; Phaff, H. J.; Miranda, M.; Miller, M. W. (1978). "Pichia cactophila, a New Species of Yeast Found in Decaying ... Phaff, H. J.; Starmer, W. T.; Miranda, M.; Miller, M. W. (1978). "Pichia heedii, a New Species of Yeast Indigenous to Necrotic ...
2002). "Functional expression of human liver cytosolic beta-glucosidase in Pichia pastoris. Insights into its role in the ...
... (formerly Pichia stipitis) is a species of yeast, belonging to the "CUG Clade" of ascomycetous yeasts ... March 2007). "Genome sequence of the lignocellulose-bioconverting and xylose-fermenting yeast Pichia stipitis". Nature ...
Pichia pastoris is a methylotrophic yeast (see Candida boidinii and Hansenula polymorpha). It provides an efficient platform ... Ogataea polymorpha (synonyms Hansenula polymorpha or Pichia angusta) is another methylotrophic yeast (see Candida boidinii). It ... Like other methylotrophic species such as Hansenula polymorpha and Pichia pastoris, it is used as a platform for the production ...
Barnett JA (1975). "The entry of D-ribose into some yeasts of the genus Pichia". Journal of General Microbiology. 90 (1): 1-12 ...
March 2007). "Genome sequence of the lignocellulose-bioconverting and xylose-fermenting yeast Pichia stipitis". Nature ... Pichia stipitis) CBS 6054, lignin/xylose degrader (2007) Spathaspora passalidarum NRRL Y-27907, model xylose fermenter (2010) ...
"A multi-level study of recombinant Pichia pastoris in different oxygen conditions". BMC Systems Biology. 4 (1): 141. doi: ...
... synthetase from Pichia ciferrii". Bioprocess Biosyst Eng. 35 (1-2): 173-81. doi:10.1007/s00449-011-0640-x. PMID 21989639. S2CID ...
"Induction and stability of alcohol oxidase in the methylotrophic yeast Pichia pastoris". Applied Microbiology and Biotechnology ...
Synthetic gene for ovine IFN-τ was produced using Pichia pastoris yeast system. The recombinant IFN-τ had the same antiviral, ...
"Production and characterization of recombinant human proteinase inhibitor 6 expressed in Pichia pastoris". Biochim. Biophys. ...
This method of expressing it in Pichia pastoris turned out to be more effective, as the produced enzyme showed cleaving ... Therefore, synthesis using Pichia pastoris seems promising for producing high quality recombinant batroxobin. As described ... In 2004, a research group from Korea produced batroxobin by expressing it in the yeast species Pichia pastoris. This ... expressed from Pichia pastoris". FEBS Letters. 571 (1-3): 67-73. doi:10.1016/j.febslet.2004.06.060. PMID 15280019. S2CID ...
Pichia is used to produce the amino acid tryptophan and the vitamin pyridoxine. Rhodotorula is used to produce the amino acid ...
Cloning, functional expression in Pichia pastoris, and substrate specificity of the isolated recombinant enzymes". The Journal ...
Yuan W, Stromhaug PE, Dunn WA (May 1999). "Glucose-induced autophagy of peroxisomes in Pichia pastoris requires a unique E1- ... Yuan W, Stromhaug PE, Dunn WA (May 1999). "Glucose-induced autophagy of peroxisomes in Pichia pastoris requires a unique E1- ... ATG7 was identified based on homology to yeast cells Pichia pastoris GSA7 and Saccharomyces cerevisiae APG7. The protein ...
Pichia Species Archived 2014-07-02 at the Wayback Machine, Doctor Fungus, url accessed 2014-02-27 Pichia at Dr Fungus Pichia in ... Some Pichia representatives can be found in raw milk and cheese, such as P. anomala (formerly named Hansenula anomala). P. ... Some Pichia species (e.g. P. ohmeri) have recently been clinically proven to be pathogens, better known as so-called ... Pichia (Hansenula and Hyphopichia are obsolete synonyms) is a genus of yeasts in the family Saccharomycetaceae with spherical, ...
Pichia heedii is a species of yeast in the family Saccharomycetaceae. Described in 1978, it was found growing on a dead senita ... Phaff HJ, Starmer WT, Miranda M, Miller MW (1978). "Pichia heedii, a new species of yeast indigenous to necrotic cacti in the ... Pichia heedii in MycoBank. v t e. ...
... Dean R. Appling appling at UTBC01.CM.UTEXAS.EDU Tue Nov 30 11:58:16 EST 1993 *Previous message: ... Several months back, a couple of postings about the Invitrogen Pichia pastoris expression system appeared. A look at the ...
Background Pichia pastoris has emerged as an important alternative host for producing recombinant biopharmaceuticals, owing to ... Despite its demonstrated utility, relatively little strain engineering has been performed to improve Pichia, due in part to the ... Comparative genomics and transcriptomics of Pichia pastoris. dc.contributor.author. Priest, Margaret. ... "Comparative Genomics and Transcriptomics of Pichia Pastoris." BMC Genomics 17.1 (2016): n. pag.. en_US. ...
Pichia guilliermondii (Candida guilliermondii anamorph; Candida melibiosi, Endomyces guilliermondii) Classification: Ascomycete ... Genus/species (aliases): Pichia guilliermondii (Candida guilliermondii anamorph; Candida melibiosi, Endomyces guilliermondii) ...
amine oxidase (copper-containing) (EC 1.4.3.6) - yeast (Pichia angusta) (fragmen... amine oxidase (copper-containing) (EC 1.4. ... amine oxidase (copper-containing) (EC 1.4.3.6) - yeast (Pichia angusta) (fragment). PIR: A38081 ...
Pichia kudriavzevii is the teleomorph of the Candida krusei. A teleomorph is the sexual reproduction stage of an organisms, ... Outbreak of Pichia kudriavzevii fungaemia in a Neonatal Intensive Care Unit. Journal of Medical Microbiology. 66: 1759-1764. ... Pichia kudriavzeii can remain metabolically active at temperatures as high as 45°C and in pHs as low as 2 (3 and 4). It ... P. kudriavzevii can be found in soil and on the outside of fruits and vegetables, often in the presence of other Pichia species ...
Kit of control elements and fluorescent reports for protein expression and secretion in Pichia pastoris from Volker Siebers ... MoClo Pichia toolkit - #1000000108. Resistance Color Key. Each circle corresponds to a specific antibiotic resistance in the ... A Modular Toolkit for Generating Pichia pastoris Secretion Libraries Obst U, Lu TK, Sieber V. ACS Synthetic Biology. 2017 May 2 ... A Modular Toolkit for Generating Pichia pastoris Secretion Libraries Obst U, Lu TK, Sieber V. ACS Synthetic Biology. 2017 May 2 ...
Production of protein-based polymers in Pichia pastoris. Promovendus. mr. ing. MWT (Marc) Werten. ... In this thesis, we explored the use of an alternative platform, namely the yeast Pichia pastoris (Komagataella phafii). We ...
Pichia pastorishas been used extensively and successfully to express recombinant proteins. In this review, we summarize the ... Pichia pastoris Protein expression Methanol induction Dissolved oxygen Gene integration Alcohol oxidase promoter AOX1 ... Pichia pastoris has been used extensively and successfully to express recombinant proteins. In this review, we summarize the ...
Neutral protease I from Aspergillus oryzae 3.042 was expressed in Pichia pastoris and its N-glycosylation properties were ... Aspergillus oryzae N-glycosylation Nano LC-MS/MS Pichia pastoris Protease Recombinant neutral protease I ... Neutral protease I from Aspergillus oryzae 3.042 was expressed in Pichia pastoris and its N-glycosylation properties were ... Glycosylation analysis of recombinant neutral protease I from Aspergillus oryzae expressed in Pichia pastoris ...
Pichia,br> Species: Membranifaciens[[#References ,[1]]] ,br> Pichia Membranifaciens originates from the genus Pichia, which ... Some of these killer yeasts include Pichia jadinii, Kluyveromyces lactis, and Pichia anomala, all of which are ... Microbiology Chromosomal differentiation of the sibling species Pichia membranifaciens and Pichia manshurica. Microbiology 78: ... Pichia genus, is capable of secreting toxins which kill other yeasts species that are sensitive to these toxins. Pichia ...
Granulocyte Macrophage Colony Stimulating Factor Human Recombinant produced in Yeast is a single, glycosylated, polypeptide chain containing 127 amino acids and having a molecular mass of 26-32 kDa. rhGMCSF differs from the natural human GM-CSF by a substitution of leucine at position 23 (R to L), and the carbohydrate moiety may be different from the native protein.
Epidermal Growth Factor Human Recombinant produced in Pichia Pastoris is a single, glycosylated, polypeptide chain containing ... Epidermal Growth Factor Human Recombinant produced in Pichia Pastoris is a single, glycosylated, polypeptide chain containing ...
Transformation efficiencies for Pichia pastoris are usually several orders of magnitude below those for other yeast. We report ... High efficiency transformation by electroporation of Pichia pastoris pretreated with lithium acetate and dithiothreitol.. Wu S1 ...
Pichia sorbitophila journal, December 2000 * de Montigny, Jacky; Spehner, Catherine; Souciet, Jean-Luc ... Heterologous protein expression in the methylotrophic yeast Pichia pastoris journal, January 2000 * Cereghino, Joan Lin; Cregg ... Optimization of humanized IgGs in glycoengineered Pichia pastoris journal, January 2006 * Li, Huijuan; Sethuraman, Natarajan; ... A strong nitrogen source-regulated promoter for controlled expression of foreign genes in the yeast Pichia pastoris journal, ...
Background: Pichia pastoris has emerged as an important alternative host for producing recombinant biopharmaceuticals, owing to ... Despite its demonstrated utility, relatively little strain engineering has been performed to improve Pichia, due in part to the ... the two strains co-branded as Pichia, has generated confusion about host performance for these genetically distinct species. ... comparative analysis of the genomic features of the three most commonly used strains comprising the tradename Pichia: K. ...
... Author(s). Chan, Joyce, M. Eng. ...
Staff publications is the digital repository of Wageningen University & Research. Staff publications contains references to publications authored by Wageningen University staff from 1976 onward.. Publications authored by the staff of the Research Institutes are available from 1995 onwards.. Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.. We have a manual that explains all the features ...
Figure 4: Production of recombinant oFGH and nFGH in Pichia pastoris. The yeast was cultured for 72 hrs with induction each 12 ... Effect of Codon Optimisation on the Production of Recombinant Fish Growth Hormone in Pichia pastoris. Hussin A. Rothan,1 Teh ... M. Romanos, "Advances in the use of Pichia pastoris for high-level gene expression," Current Opinion in Biotechnology, vol. 6, ... In this study, a significant improvement in yield and function of recombinant FGH was achieved in Pichia pastoris using a ...
However, the adaptability of Pichia stipitis CBS 5776 in the prehydrolyzate was so limited that steam strip-ping and overliming ... and then the acid hydrolyzate was detoxified and fermented by Pichia stipitis CBS 5776. Steam stripping could remove volatile ... Yeast Pichia stipitis CBS 5776 was developed through adaptation fermentation step by step in steam exploded corn stover ... "Adaptation fermentation of Pichia stipitis and combination detoxification on steam exploded lignocellulosic prehydrolyzate" ...
X-ray crystallography and mass spectroscopy reveal that the N-lobe of human transferrin expressed in Pichia pastoris is folded ... HUMAN SERUM TRANSFERRIN, N-TERMINAL LOBE, EXPRESSED IN PICHIA PASTORIS. *DOI: 10.2210/pdb1b3e/pdb ...
The recombinant lipase was expressed in Pichia pastoris GS115 cells. The recombinant lipase was found to have a molecular mass ... Cloning and Expression of Aspergillus tamarii FS132 Lipase Gene in Pichia pastoris Bihong Shi 1,2,* , Liqing Zeng 1. ... Keywords: Aspergillus tamarii; lipase; gene cloning; expression; Pichia pastoris Aspergillus tamarii; lipase; gene cloning; ... Shi B, Zeng L, Song H, Shi Q, Wu S. Cloning and Expression of Aspergillus tamarii FS132 Lipase Gene in Pichia pastoris . ...
In Scheffersomyces (Pichia) stipitis and related fungal species the genes for L-rhamnose catabolism RHA1, LRA2, LRA3 and LRA4 ... Pichia stipitis genomics, transcriptomics, and gene clusters. *Whole-transcriptome response to water stress in a California ... Characterisation of the gene cluster for L-rhamnose catabolism in the yeast Scheffersomyces (Pichia) stipitis ... Characterisation of the gene cluster for L-rhamnose catabolism in the yeast Scheffersomyces (Pichia) stipitis. Gene. 492: 177- ...
The methylotrophic yeast Pichia pastoris is widely used in the manufacture of industrial enzymes and pharmaceuticals. Like most ... nov. and the transfer of Pichia pseudopastoris to the methylotrophic yeast genus Komagataella. Int. J. Syst. Evol. Microbiol. ... Supplementary Figure 1 Strategy for engineering chemoorganoautotrophy in Pichia pastoris.. (a) Wild type P. pastoris is able to ... Pichia pastoris: protein production host and model organism for biomedical research. Future Microbiol. 8, 191-208 (2013). ...
Use the Pichia secreted expression system - White ppt formed in the supernatant after stored at -80 C fridge (Mar/12/2006 ). ... I´ve used the same expression method and Pichia also secrete a lot of native proteins. I didn´t saw white ppt, but I did my ... I´ve used the same expression method and Pichia also secrete a lot of native proteins. I didn´t saw white ppt, but I did my ... I m working on the Pichia expression system. I cloned my target gene into the pPICZ-alpha-A vector(the recombinant protein will ...
... expressed in Pichia pastoris; find Roche-04716728001 MSDS, related peer-reviewed papers, technical documents, similar products ... from bovine pancreas, expressed in Pichia pastoris * Enzyme Commission (EC) Number 3.1.21.1 ( BRENDA. , IUBMB. ) ...
We report on an efficient ultrasound based technique for lysing Escherichia coli and Pichia pastoris with oscillating ... Sonolysis of Escherichia coli and Pichia pastoris in microfluidics Tandiono Tandiono,*a Dave Siak-Wei Ow,b Leonie Driessen,c ... Sonolysis of Escherichia coli and Pichia pastoris in microfluidics T. Tandiono, D. Siak-Wei Ow, L. Driessen, C. Sze-Hui Chin, E ... We report on an efficient ultrasound based technique for lysing Escherichia coli and Pichia pastoris with oscillating ...
Inducible high-level expression is favoured for recombinant protein production in Pichia pastoris. Therefore, novel regulated ... Ahn J,Hong J,Park M,Lee H,Lee E,Kim C,Lee J,Choi E,Jung J,Phosphate-responsive promoter of a Pichia pastoris sodium phosphate ... Keywords: Pichia pastoris, Heterologous protein production, Glucose-limited fed batch cultivation, Inducible promoter, High- ... Ahn J,Hong J,Lee H,Park M,Lee E,Kim C,Choi E,Jung J,Translation elongation factor 1-alpha gene from Pichia pastoris: molecular ...
  • Grinna LS, Tschopp JF (1989) Size distribution and general structural features of N-linked oligosaccharides from the methylotrophic yeast, Pichia pastoris . (springer.com)
  • Montesino R, Garcia R, Quintero O et al (1998) Variation in N-linked oligosaccharide structures on heterologous proteins secreted by the methylotrophic yeast Pichia pastoris . (springer.com)
  • Expression of human collagen in the methylotrophic yeast Pichia pastoris. (wur.nl)
  • The methylotrophic yeast Pichia pastoris is widely used in the manufacture of industrial enzymes and pharmaceuticals. (nature.com)
  • To improve ARA production with engineered methylotrophic yeast Pichia pastoris , a genetically modified ara gene from Aspergillus niger ND-1 was investigated. (frontiersin.org)
  • The methylotrophic yeast Pichia pastoris is an attractive expression host due to its ease of manipulation and its capacity to perform post-translational protein modifications, such as N-glycosylation [Daly and Hearn (2005) J Mol Recognit 18: 119-138]. (diva-portal.org)
  • TY - CHAP UR - http://lib.ugent.be/catalog/pug01:3129907 ID - pug01:3129907 LA - eng TI - Production of antibody derivatives in the methylotrophic yeast Pichia pastoris PY - 2012 SN - 9781617799747 SN - 9781617799730 SN - 1064-3745 PB - Clifton AU - Schoonooghe, Steve UGent 001994270082 AU - Leoen, Jannick AU - Haustraete, Jurgen AU - Chames, Patrick editor AB - New antibody derivatives are continuously being generated to interact with a range of therapeutic targets. (ugent.be)
  • The cDNA encoding a xyloglucan endotransglycosylase, PttXET16A, from hybrid aspen ( Populus tremula × tremuloides ) has been isolated from an expressed sequence tag library and expressed in the methylotrophic yeast Pichia pastoris . (portlandpress.com)
  • EC 1.6.6.1) was produced in the methylotrophic yeast Pichia pastoris and purified to near-electrophoretic homogeneity. (plantphysiol.org)
  • The methylotrophic, Crabtree-negative yeast Pichia pastoris is widely used as a heterologous protein production host. (biomedcentral.com)
  • Blanchard V, Gadkari RA, Gerwig GJ, Leeflang BR, Dighe RR, Kamerling JP (2007) Characterization of the N-linked oligosaccharides from human chorionic gonadotropin expressed in the methylotrophic yeast Pichia pastoris . (springer.com)
  • Sambuk, E. 2013-04-21 00:00:00 Pichia pastoris is a methylotrophic yeast that is widely used for the expression of heterologous proteins. (deepdyve.com)
  • The methylotrophic yeast Pichia pastoris can be used to express recombinant genes at high levels under the control of the methanol-inducible alcohol oxidase 1 (AOX1) promoter. (ubc.ca)
  • Expression and transport characterisation of the wheat low-affinity cation transporter (LCT1) in the methylotrophic yeast Pichia pastoris. (lancs.ac.uk)
  • The low-affinity cation transporter (LCT1) from wheat (Triticum aestivum) was expressed in the methylotrophic yeast Pichia pastoris and its transport characteristics studied employing Ca45 and Cd109. (lancs.ac.uk)
  • A 2526 bp gene encoding Aspergillus niger Beta-glucosidase was chemically synthesized for its heterologous expression in methylotrophic yeast Pichia pastoris , using methanol as inducer. (ccsenet.org)
  • The methylotrophic yeast Pichia pastoris is firmly established as a host for the production of recombinant proteins, frequently outperforming other heterologous hosts. (uni-bielefeld.de)
  • Genome-scale metabolic model of methylotrophic yeast Pichia pa. (mysciencework.com)
  • Genome-scale metabolic model of methylotrophic yeast Pichia pastoris and its use for in silico analysis of heterologous protein production. (mysciencework.com)
  • The methylotrophic yeast Pichia pastoris has gained much attention during the last decade as a platform for producing heterologous recombinant proteins of pharmaceutical importance, due to its ability to reproduce post-translational modification similar to higher eukaryotes. (mysciencework.com)
  • The aim of the present study was to express TPL in the methylotrophic yeast Pichia pastoris in order to get a large amount of this enzyme exhibiting interesting biochemical properties, to purify and characterize the recombinant enzyme. (biomedcentral.com)
  • Methylotrophic yeast Pichia pastoris is an object of modern biotechnology. (scirp.org)
  • In Scheffersomyces ( Pichia ) stipitis and related fungal species the genes for L -rhamnose catabolism RHA1, LRA2, LRA3 and LRA4 but not LADH are clustered. (usda.gov)
  • Water-hyacinth's (Eichhornia crassipes) hemicellulose acid hydrolysate has been utilized as a substrate for alcohol production using Pichia stipitis NCIM3497. (environmental-expert.com)
  • Cloning novel sugar transporters from Scheffersomyces (Pichia) stipitis allowing D-xylose fermentation by recombinant Saccharomyces cerevisiae. (sigmaaldrich.com)
  • The groundnut shell enzymatic hydrolysate (45.6 g/L reducing sugars) was fermented for ethanol production with free and sorghum stalks immobilized cells of Pichia stipitis NCIM 3498 under submerged cultivation conditions. (degruyter.com)
  • In Pichia stipitis , fermentative and pyruvate decarboxylase (PDC) activities increase with diminished oxygen rather than in response to fermentable sugars. (asm.org)
  • Pichia stipitis is one of the best-known xylose-fermenting yeasts ( 5 ). (asm.org)
  • Xylose reductase (XR) and xylitol dehydrogenase (XDH) from Pichia stipitis are the two enzymes most commonly used in recombinant Saccharomyces cerevisiae strains engineered for xylose utilization. (biomedcentral.com)
  • However, in a recent study, a strain carrying the Pichia stipitis XR-XDH pathway showed significantly higher xylose consumption rate and higher specific ethanol productivity compared with an isogenic strain carrying the Pyromyces XI pathway [ 12 ]. (biomedcentral.com)
  • Fish growth hormone has been expressed in the yeast Saccharomyces cerevisiae [ 8 - 10 ] and the yeast Pichia pastoris [ 11 ]. (hindawi.com)
  • Molecular phylogenetic analyses showed that the various species of Pichia were widely scattered within the Saccharomycotina, a fungal clade that includes a large number of yeast species (including such familiar taxa as the brewer's or baker's yeast Saccharomyces cerevisiae ). (fieldofscience.com)
  • Biocontrol of Mold Growth in High-Moisture Wheat Stored under Airtight Conditions by Pichia anomala, Pichia guilliermondii, and Saccharomyces cerevisiae. (diva-portal.org)
  • Die swak sekresie en komplekse proteïenstruktuur van lignienperoksidase het tot dusver die produksie daarvan in natuurlike gashere sowel as in heteroloë uitdrukkingstelsels soos Escherichia coli en Saccharomyces cerevisiae beperk. (sun.ac.za)
  • Induction without methanol: novel regulated promoters enable high-level expression in Pichia pastoris. (biomedsearch.com)
  • Day 2 : Methanol-free expression of SARS-CoV2-RBD variants in Pichia pastoris. (eventinterface.com)
  • Two methanol feeding methods, namely constant methanol feed and on-line monitoring feed control by methanol sensor were investigated to improve the production of recombinant human growth hormone (rhGH) in high cell density cultivation of Pichia pastoris KM71 in 2 L bioreactor. (ajol.info)
  • Pichia pistoris is a type of yeast that feed on methanol and does not normally produce α-amylase enzyme. (avroarrow.org)
  • Therefore, the main aim of this work was to optimize the temperature, dimethylsulfoxide (DMSO) concentration and the methanol flow-rate for the biosynthesis of recombinant MBCOMT by Pichia pastoris bioreactor methanol-induced cultures using artificial neural networks (ANN). (biomedcentral.com)
  • Scaling-up of traditional Pichia fermentation processes can be challenging due to use of methanol and complex feed regimes. (technologynetworks.com)
  • This collection of control elements and fluorescent reporters can be used in combination with the MoClo-YTK toolkit for protein expression and secretion in Pichia pastoris . (addgene.org)
  • The Sieber Lab developed the MoClo Pichia Toolkit as a set of genetic parts to control transcription, translation, and protein secretion in P. pastoris . (addgene.org)
  • Pichia pastoris protein secretion platform. (addgene.org)
  • Targeting this protein for secretion in Pichia pastoris results in its retention within the cell. (cornell.edu)
  • Enhancement of Protein Secretion in Pichia pastoris by Overexpression " by Mehmet Inan, Dinesh Aryasomayajula et al. (unl.edu)
  • Since the protein was glycosylated during secretion from Pichia pastoris , therefore, the role of carbohydrate moieties on its stability was analyzed via in vivo blocking of N-glycosylation using tunicamycin where an increased degradation of streptokinase was observed. (springer.com)
  • We discovered, surprisingly, that the secretion, processing, and function of an AcbA-derived peptide, SDF-2, are conserved between the yeast Pichia pastoris and D. discoideum . (rupress.org)
  • In this study, we define the novel pathways and proteins necessary for AcbA-like protein (Acb1 in Pichia pastoris ) secretion in yeast. (rupress.org)
  • Day 3 : Silk secretion in Pichia. (eventinterface.com)
  • ABSTRACT: BACKGROUND: Inducible high-level expression is favoured for recombinant protein production in Pichia pastoris. (biomedsearch.com)
  • An abstract of the paper "Cloning and Overexpression of a Yeast Phytase Gene in Pichia Pastoris," by Virginie Neugnot-Roux, Guy Moulin, and colleagues, is presented. (ebscohost.com)
  • Cloning and overexpression of a yeast phytase gene in Pichia pastoris. (ebscohost.com)
  • MEY-1: gene cloning and overexpression in Pichia pastoris. (ebscohost.com)
  • Overexpression of the S. cerevisiae GSHI and GSHII genes in S. cerevisiae ( 5 , 20 ) and Pichia pastoris ( 6 ) has also been reported. (asm.org)
  • After identification of the sequence encoding active Hac1p we evaluated the effect of its overexpression in Pichia. (harvard.edu)
  • RSM optimization of HSA/IL1Ra in Pichia pastoris overexpression strain and study of its in vivo activity in reducing hyperglycemia of GK rats. (nextbio.com)
  • Shi B, Zeng L, Song H, Shi Q, Wu S. Cloning and Expression of Aspergillus tamarii FS132 Lipase Gene in Pichia pastoris . (mdpi.com)
  • Both gene products were expressed in Pichia pastoris. (nih.gov)
  • 2. It is absolutely mandatory to verify the correct integration of your (linarized) gene into the Pichia genome! (protocol-online.org)
  • When U linearize in the AOX1-promoter site, the gene is recombinated into the Pichia AOX promoter site. (protocol-online.org)
  • Results: The mannan endo-1,4-β-mannosidase gene ( MAN) from Aspergillus niger CBS 513.88 was optimized based on the codon usage bias in Pichia pastoris and. (ebscohost.com)
  • The β-mannanase gene (1,029 nucleotide) from Bacillus subtilis MAFIC-S11, encoding a polypeptide of 342 amino acids, was cloned and expressed in Pichia pastoris. (ebscohost.com)
  • De novo synthesis, constitutive expression of Aspergillus sulphureus β-xylanase gene in Pichia pastoris and partial enzymic characterization. (ebscohost.com)
  • The endo-β-1, 4-xylanase gene xynA from Aspergillus sulphureus, encoded a lack-of-signal peptide protein of 184 amino acids, was de novo synthesized by splicing overlap extension polymerase chain reaction according to Pichia pastoris protein’s codon bias. (ebscohost.com)
  • The sequence of an endo-chitosanase gene ( CSN) from Aspergillus fumigatus was optimized based on the preferred codons of Pichia pastoris and synthesized in vitro through overlapping PCR ( CSN- P). The gene was cloned into a yeast expression vector, pHBM905A, and secretorily expressed in Pichia. (ebscohost.com)
  • In this report, we describe a modified concatemerization strategy to construct a gene with enhanced sequence diversity that encodes a highly repetitive elastin-like protein polymer for expression in Pichia pastoris . (pubmedcentralcanada.ca)
  • Abad S, Kitz K, Hormann A, Schreiner U, Hartner FS, Glieder A (2010) Real-time PCR-based determination of gene copy numbers in Pichia pastoris . (springer.com)
  • Pichia pastoris was transformed with an integrative plasmid containing the JEN1 gene. (biochemj.org)
  • Day 4 : Improvement of bioprocess efficiency in Pichia pastoris recombinant protein production combining gene dosage and oxygen limitation. (eventinterface.com)
  • Oxford Gene Technology (OGT) and Integrated Genomics (IG) have announced the signing of a collaboration to develop and market a Pichia pastoris Gene Expression (GE) oligonucleotide microarray. (technologynetworks.com)
  • To further such studies in the biotechnology favored yeast Pichia pastoris, we cloned and characterized the P. pastoris HAC1 gene and the splice event. (harvard.edu)
  • Expression of α-amylase gene in Escherichia coli and Pichia pastoris. (avroarrow.org)
  • Hence by isolating and expressing the specific gene that responsible for the production of α-amylase enzyme into another microorganism that easier to grow such as E. coli and Pichia pastoris will not only reduce the cost but also ease the enzyme purification (Nimkar et al. (avroarrow.org)
  • The gene was fused to the pPICZαA plasmid and overexpressed in Pichia pastoris SMD1168. (dtu.dk)
  • To compare the DAAO expression level in different Pichia pastoris host strains, the gene encoding DAAO from Trigonopsis variabilis was cloned into plasmid pPIC3.5k and then transformed into P. pastoris GS115 and KM71 respectively. (semanticscholar.org)
  • Guo M, Hang H, Zhu T et al (2008) Effect of glycosylation on biochemical characterization of recombinant phytase expressed in Pichia pastoris . (springer.com)
  • High-level expression and characterization of a thermophilic β-mannanase from Aspergillus niger in Pichia pastoris. (ebscohost.com)
  • Cloning, Expression, and Characterization of β-mannanase from Bacillus subtilis MAFIC-S11 in Pichia pastoris. (ebscohost.com)
  • High-level expression and characterization of a highly thermostable chitosanase from Aspergillus fumigatus in Pichia pastoris. (ebscohost.com)
  • Day 1 : Expression and characterization of biologically active glycoprotein hormones using the Pichia GlycoSwitch® technology. (eventinterface.com)
  • Here we describe the production, purification and characterization of Bacillus licheniformis chitinase A in the Pichia pastoris expression system. (iupui.edu)
  • Furthermore, the co-mingling of genomic, transcriptomic and fermentation data collected about Komagataella pastoris and Komagataella phaffii, the two strains co-branded as Pichia, has generated confusion about host performance for these genetically distinct species. (mit.edu)
  • In this thesis, we explored the use of an alternative platform, namely the yeast Pichia pastoris (Komagataella phafii). (wur.nl)
  • Somewhat unfortunately, one of the species to be expelled from Pichia is perhaps the best-studied: the yeast formerly known as Pichia pastoris (now supposed to be referred to as Komagataella pastoris though a quick Google Scholar search suggests that a great many authors are pretending that hasn't happened). (fieldofscience.com)
  • The study describes the identification of sphingolipid biosynthesis genes in the non-conventional yeast Pichia ciferrii, the development of tools for its genetic modification as well as their application for metabolic engineering of P. ciferrii with the goal to generate strains capable of producing the rare sphingoid bases sphinganine and sphingosine. (nih.gov)
  • Mechanism of transformation in Pichia methanolica yeast: transforming and nontransforming genes]. (semanticscholar.org)
  • Two types of genes were found in the study of transformation in yeast Pichia methanolica: transforming (Trg) and nontransforming (Ntg) genes. (semanticscholar.org)
  • Neutral protease I from Aspergillus oryzae 3.042 was expressed in Pichia pastoris and its N -glycosylation properties were analyzed. (springer.com)
  • Ke Y, Huang WQ, Li JZ et al (2012) Enzymatic characteristics of a recombinant neutral protease I (rNpI) from Aspergillus oryzae expressed in Pichia pastoris . (springer.com)
  • In this study we report the heterologous expression of Aspergillus oryzae β-xylosidase (XylA) in Pichia pastoris under the control of the glyceraldehyde-3-phosphate dehydrogenase promoter. (nottingham.ac.uk)
  • The budding yeast Pichia pastoris responds to methanolic media by synthesizing high levels of cytosolic enzymes (e.g. formate dehydrogenase) and peroxisomal enzymes (e.g. alcohol oxidase), which are necessary to assimilate this carbon source. (biologists.org)
  • The invention relates to novel cytotoxic proteins produced by the yeast Pichia inositovora. (google.com)
  • Here, we describe the construction of the expression vectors needed for heterologous expression of a Fab fragment in the yeast Pichia pastoris. (ugent.be)
  • Trimeresurus stejnegeri lectin (TSL) was expressed in the yeast Pichia pastoris under the control of alcohol oxidase (AOX1) promoter. (eurekaselect.com)
  • The data gathered were modelled via PARAFAC-PLS chemometric methodologies, resulting in important qualitative and quantitative information about the behaviours of different biogenic fluorophors in batch cultures of the yeast Pichia pastoris. (ebscohost.com)
  • We previously described the isolation of mutants of the yeast Pichia pastoris that are deficient in peroxisome assembly (pas mutants). (rupress.org)
  • Recombinant SFP2 was expressed in and secreted from the yeast Pichia pastoris with a final yield of 78 mg/L (136.2 U/mL caseinolytic activity) after 25 h of induction. (eurekamag.com)
  • The yeast Pichia pastoris is a good alternative host for heterologous protein expression and has been used to produce many enzymes for industrial applications due to its high recombinant protein titres, inexpensive cultivation methods, Generally Regarded As Safe (GRAS) status and clear-cut downstream processes. (sun.ac.za)
  • The yeast Pichia pastoris (syn. (biomedcentral.com)
  • Genetic Control of Purine Biosynthesis in Yeast Pichia methanolica. (semanticscholar.org)
  • Despite its demonstrated utility, relatively little strain engineering has been performed to improve Pichia, due in part to the limited number and inconsistent frameworks of reported genomes and transcriptomes. (mit.edu)
  • The protein is produced by culture of Pichia inositovora strain NRRL Y-18709, and may be subsequently recovered from the culture medium and purified. (google.com)
  • A unique resistant strain was selected and identified as Pichia anomala (Wickerhamomyces anomalus), deposited as CBS 132101. (metabolomicscentre.nl)
  • State-of-the-art strain engineering techniques for the host Pichia pastoris (syn. (biomedcentral.com)
  • Background Pichia pastoris has emerged as an important alternative host for producing recombinant biopharmaceuticals, owing to its high cultivation density, low host cell protein burden, and the development of strains with humanized glycosylation. (mit.edu)
  • Results Here, we present a comprehensive and standardized comparative analysis of the genomic features of the three most commonly used strains comprising the tradename Pichia: K. pastoris wild-type, K. phaffii wild-type, and K. phaffii GS115. (mit.edu)
  • The behavior of Pichia guilliermondii strains producing high levels of 4-ethylphenol in synthetic media was studied in wines and grape juices. (ajevonline.org)
  • Compared D-amino acid oxidase expression in different Pichia pastoris host strains]. (semanticscholar.org)
  • We report on an efficient ultrasound based technique for lysing Escherichia coli and Pichia pastoris with oscillating cavitation bubbles in an integrated microfluidic system. (rsc.org)
  • Higher levels of the target proteins were achieved by intracellular and extracellular heterologous production using an Escherichia coli and Pichia pastoris based expression system respectively. (dissertations.se)
  • Several months back, a couple of postings about the Invitrogen Pichia pastoris expression system appeared. (bio.net)
  • Dear All, I m working on the Pichia expression system. (protocol-online.org)
  • Day 1 : Introduction to the Pichia pastoris Expression System. (eventinterface.com)
  • The combination of OGT's oligonucleotide array expertise combined with Integrated Genomics sequence and detailed Pichia genome annotations will be a powerful molecular research tool for the Pichia community and add strength to our growing portfolio. (technologynetworks.com)
  • Pichia cells had earlier been transformed with the constructs and the transgene integrated into Pichia genome by homologous recombination. (slu.se)
  • Pichia (Hansenula and Hyphopichia are obsolete synonyms) is a genus of yeasts in the family Saccharomycetaceae with spherical, elliptical, or oblong acuminate cells. (wikipedia.org)
  • Pichia Membranifaciens'' originates from the genus Pichia, which previously was a polyphyletic group based largely on ascospore morphology. (kenyon.edu)
  • One of the harder-hit taxa was the genus Pichia , previously recognised as a large genus of close to 100 species. (fieldofscience.com)
  • As a result, the genus has been progressively pared down to a smaller array of species concentrated around the type, Pichia membranifaciens . (fieldofscience.com)
  • The performed experiments concerned expression of myrosinases in Pichia pastoris aiming to test their functionality based on recombinant enzyme activity and to compare different expression and cell breakage systems. (slu.se)
  • The anamorphs of some Pichia species are Candida species. (wikipedia.org)
  • The invention provides orthogonal translation systems for the production of polypeptides comprising unnatural amino acids in methyltrophic yeast such as Pichia pastoris. (osti.gov)
  • Researchers at New Brunswick Scientific (Edison, NJ) found that by switching from a shaker to a fermentor, protein production in Pichia could be increased by over 140% (3). (bio-medicine.org)
  • Developments in process techniques for production and recovery of heterologous proteins with Pichia pastoris are presented. (diva-portal.org)
  • Parallelized microplate cultivation of the eukaryotic protein production host Pichia pastoris was applied effectively to support miniaturized phenotyping of clonal libraries in batch as well as fed-batch mode. (altmetric.com)
  • Thus, the effect of osmolarity on the cellular physiology of Pichia pastoris , a prominent host for recombinant protein production, was studied in carbon limited chemostat cultures at different osmolarities. (biomedcentral.com)
  • Adivitiya, Dagar VK, Devi N, Khasa YP (2016) High level production of active streptokinase in Pichia pastoris fed-batch culture. (springer.com)
  • Ayed A, Rabhi I, Dellagi K, Kallel H (2008) High level production and purification of human interferon α2b in high cell density culture of Pichia pastoris . (springer.com)
  • Day 1 : Bioprocess development through AI Models to control cell physiology in Pichia pastoris recombinant protein production. (eventinterface.com)
  • Day 3 : Establishing Pichia pastoris as a promising candidate for bulk chemicals production: the 3-hydroxypropionic acid case. (eventinterface.com)
  • Day 4 : Bioprocessing with Pichia pastoris Production of recombinant pharmaceutical proteins and peptides. (eventinterface.com)
  • Compared to the highest previously reported value, this approach resulted in an ~2 fold enhancement of IP production using Pichia based expression systems, thus significantly increasing the efficiency of insulin manufacture. (pubmedcentralcanada.ca)
  • Pichia pastoris was designed as a valuable alternative to E. coli and CHO cells for production of novel protein formats such as multi-specific antibody mimetics. (technologynetworks.com)
  • Lonza has established a new XS™ Pichia 2.0 system designed to provide high product titers (up to 6 g/L) for these novel compounds along with a fast, robust and scalable manufacturing process suitable for commercial production. (technologynetworks.com)
  • Some Pichia representatives can be found in raw milk and cheese, such as P. anomala (formerly named Hansenula anomala). (wikipedia.org)
  • A total of 17 Pichia ( Wickerhamomyces ) isolates obtained from enological ecosystems in the Utiel-Requena Spanish region were characterized by physiological (using API 20C AUX strips and ID Yeast Plus System miniaturized identification systems) and molecular (PCR-RFLP and sequencing) techniques as belonging to the species P. fermentans, P. membranifaciens , and W. anomalus . (ajevonline.org)
  • 2010), which used Pichia pastoris as host, they first digested the plasmid Blunt II-Topo that contain the fragment DNA from Bacillus subtilis as explained earlier with PmlI and KpnI before purified it. (avroarrow.org)
  • In winemaking, some species of Pichia can create potential faults in wines. (wikipedia.org)
  • Some Pichia species (e.g. (wikipedia.org)
  • Pichia heedii is a species of yeast in the family Saccharomycetaceae. (wikipedia.org)
  • P. kudriavzevii can be found in soil and on the outside of fruits and vegetables, often in the presence of other Pichia species. (kenyon.edu)
  • Epidermal Growth Factor Human Recombinant produced in Pichia Pastoris is a single, glycosylated, polypeptide chain containing 51 amino acids and having a molecular mass of 6KDa. (prospecbio.com)
  • Azoun SB, Belhaj AE, Kallel H (2016) Rabies virus glycoprotein enhanced expression in Pichia pastoris using the constitutive GAP promoter. (springer.com)
  • High efficiency transformation by electroporation of Pichia pastoris pretreated with lithium acetate and dithiothreitol. (nih.gov)
  • Transformation efficiencies for Pichia pastoris are usually several orders of magnitude below those for other yeast. (nih.gov)
  • But my transformation of pichia failed for many times. (protocol-online.org)
  • The mutagenesis products were cloned into the Pichia pastoris expression vector pPICZB for transformation of P. pastoris SMD1168. (essays.se)
  • Fermentation of Xylose: Xylose is inputted into the pentose phosphate pathway and converted into fructose (this is unique to Pichia kudriavzeii ). (kenyon.edu)
  • Day 4 : High cell density cultivations of Pichia pastoris in microbioreactor system. (eventinterface.com)
  • I had some trouble transforming Pichia, and finally managed some good transformants by increasing amount of linealized DNA used in electroporation, Pichia is tricky to transform, you should be using 1-5 ug DNA aprox. (protocol-online.org)
  • Charoenrat T, Khumruaengsri N, Promdonkoy P, Rattanaphan N, Eurwilaichitr L, Tanapongpipat S, Roongsawang N (2013) Improvement of recombinant endoglucanase produced in Pichia pastoris KM71 through the use of synthetic medium for inoculum and pH control of proteolysis. (springer.com)
  • Expression and purification of human TAT-p53 fusion protein in Pichia pastoris and its influence on HepG2 cell apoptosis. (biomedsearch.com)
  • Therefore, the P. pastoris was used because it is a yeast that has by nature a pattern of post- translational modifications of proteins similar in their patterns of mammals when comparing this genre (Pichia) with other genera of yeasts now used in heterologous expression. (ibict.br)
  • Dr Theresa Walunas, Vice President of Integrated Genomics added: "OGT's IJISS technology provides an important new capability to the Pichia research community. (technologynetworks.com)
  • Pichia is a teleomorph, and forms hat-shaped, hemispherical, or round ascospores during sexual reproduction. (wikipedia.org)
  • A teleomorph is the sexual reproduction stage of an organisms, meaning Pichia kudriavzevii reproduces sexually through the fusion of haploid cells (15). (kenyon.edu)
  • Compared to other eukary otic expression systems, Pichia offers many advantages, because it does not have the endotoxin problem associated with bacteria nor the viral contamination problem of proteins produced in animal cell culture. (bio-medicine.org)
  • Boraston AB, McLean BW, Guarna MM, Amandaron-Akow E, Kilburn DG (2001a) A family 2a carbohydrate-binding module suitable as an affinity tag for proteins produced in Pichia pastoris . (springer.com)
  • The recombinant lipase was expressed in Pichia pastoris GS115 cells. (mdpi.com)
  • Els cultius en discontinu en bioreactor van revelar l'efecte sinèrgic del metanol i la metilamina com a substrats inductors sobre el promotor FLD1. (tesisenred.net)