Quantification of Endoglucanase Activity based on Carboxymethyl Cellulose in Four Fungi Isolated from an Aerated Lagoon in a Pulp and Paper Mill. Luis A. Ordaz-Díaz,a Juan A. Rojas-Contreras,b Felipe Flores-Vichi,δc Mónica Y. Flores-Villegas,a Carlos Álvarez-Álvarez,a Pryscila Velasco-Vázquez,a and Ana M. Bailón-Salas b,*. The aim of this study was to identify cellulolytic fungal strains capable of degrading cellulose from an aerated lagoon in a pulp and paper mill. Four fungal strains that were found to be highly active were isolated on carboxymethyl cellulose (CMC) and suggested to be CMCase/endoglucanase. The identified strains were Aspergillus niger, Penicillium sp.,Aspergillus fumigatus, and Mucor sp. All the strains were studied in terms of cultural morphological characteristics and microscopic examinations. The endoglucanase with the highest isolate production was Penicillium sp., which also showed the highest qualitative endoglucanase activity (1.3 cm), in addition to the main ...
Abstract: Background and Objective: Cellulase as a fibrolytic enzyme is a highly effective tool for agricultural waste treatments. Production of cellulase enzyme on medium of agricultural wastes by Fusarium graminearum to be used in ruminant feeding was the main objective of this study. Materials and Methods: Impact of initial pH of growth medium, different nitrogen sources and variety of agriculture by products as a carbon sources on cellulase production have been studied. Electron microscope was used for investigate the impact of the resultant cellulase on corn stover degradation, while batch culture technique was used for investigate impact of different levels of the produced and commercial cellulases on total mixed ration digestibility by rumen microorganisms (in vitro). Results: Cellulase maximum production by F. graminearum was obtained at 20% corn stover, initial pH of growth medium 5.0 and peptone as a nitrogen source. All addition levels of the produced cellulase increased dry matter ...
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Direct utilization of untreated oil palm trunk (OPT) for cellulases and xylanase production by Aspergillus fumigatus SK1 was conducted under solid-state fermentation (SSF). The highest activities of extracellular cellulases and xylanases were produced at 80% moisture level, initial pH 5.0, 1 × 108 spore/g (inoculum) with 125 μm of OPT as sole carbon source. The cellulases and xylanase activities obtained were 54.27, 3.36, 4.54 and 418.70 U/g substrates for endoglucanase (CMCase), exoglucanase (FPase), β-glucosidase and xylanase respectively. The crude cellulases and xylanase required acidic condition to retain their optimum activities (pH 4.0). Crude cellulases and xylanase were more stable at 40°C compared to their optimum activities conditions (60°C for FPase and 70°C for CMCase, β-glucosidase and xylanase). SDS-PAGE and zymogram analysis showed that Aspergillus fumigatus SK1 could secrete cellulases (endoglucanase, exoglucanase and β-glucosidase), xylanase and protease. Enzymatic ...
Results: It was observed that cellulase activity of T. reesei strains varies from 0.73 to 3.11 IU and T. reesei 5A shown maximum cellulase activity after 96 h of growth perod at 37 °C. Total genomic DNA was isolated from T. reesei 5A and then it was partially digested with Sau 3A and ligated to vector YEpFLAG-1, linearized with BamH1. The construct was used to transform Escherichia coli and recombinant clone(s) were screened on Reese medium supplemented with carboxymethyl cellulose and ampicillin. The E. coli recombinant clones were further confirmed by gene specific amplification using PCR. S. cerevisiae was transformed with the recombinant plasmids YEpFLAG-1-cel-exo and YEpFLAG-1-cel-endo isolated from E. coli transformants. Carboxymethyl cellulase (CMCase) activity was observed in three yeast transformants Tr-2, Tr-4 and Tr-6 as 0.50, 0.70 and 0.80 IU, respectively. no FPase (exoglucanse) activity was observed in any of the yeast transformants ...
Allcosmeticsource.com Cellulase 100,000u/g,1kg/bag,free shipping [EP170508014]- Cellulase 100,000u/g,1kg/bag,free shipping What is Cellulase 100,000u/g Cellulase refers to the total name of multiple enzymes which can catalyze and hydrolyze cellulose. Generally, cellulase which can hydrolyze natural cellulose contains three activity constituents: endoglucanase, exoglucanase and glucuroide. Function of Cellulase 100,000u/g (1) The effect of snowflake, stereoscopic and color brightness can be further improved (2)
Disclosed are improved methods for treating cotton-containing fabrics as well as the fabrics produced from these methods. In particular, the disclosed methods are directed to contacting cotton-containing fabrics with a cellulase solution containing a fungal cellulase composition which is substantially,free of all CBH I type cellulase components. Cotton-containing fabrics so treated possess decreased strength loss as compared to fabrics treated with a cellulase solution containing a complete cellulase composition.
Deletion of Cel48S from C. thermocellum led to a decrease in the enzymatic hydrolysis rate, a decrease in microbial hydrolysis rate, and a decrease in biomass formation during growth on Avicel.. The similarity of enzyme saturation curves for the WT and parent strains suggests that the ΔpyrF mutation in the parent strain has no effect on cellulosome function, as expected. The S mutant strain, however, exhibited a reduction in both specific activity and saturation rate. A reduction in specific activity is indicative of impaired function and consistent with decreased synergy among components of the cellulosome in the absence of Cel48S (3).. The role of GH families in cellulose solubilization is a topic of much debate. Family 48 cellulases are a prominent component of many bacterial cellulase systems and, due to their ubiquity, are thought to play an important role in cellulose solubilization (21). On one hand, disruption of the single family 9 GH in C. phytofermentans eliminated its ability to ...
Cellulases have numerous applications in several industries, including biofuel production, food and feed industry, brewing, pulp and paper, textile, laundry, and agriculture.Cellulose-degrading bacteria are widely spread in nature, being isolated from quite different environments. Cellulose degradation is the result of a synergic process between an endoglucanase, an exoglucanase and a,β-glucosidase. Bacterial endoglucanases degrade ß-1,4-glucan linkages of cellulose amorphous zones, meanwhile exoglucanases cleave the remaining oligosaccharide chains, originating cellobiose, which is hydrolyzed by ß-glucanases. Bacterial cellulases (EC 3.2.1.4) are comprised in fourteen Glycosil Hydrolase families. Several advantages, such as higher growth rates and genetic versatility, emphasize the suitability and advantages of bacterial cellulases over other sources for this group of enzymes. This review summarizes the main known cellulolytic bacteria and the best strategies to optimize their cellulase
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A novel hemicellulase-producing fungal strain was isolated from a local soil sample. The organism is identified as Aspergillus fumigatus based on ribosomal RNA analyses. The Aspergillus strain, designated as 2NB, produces both enzymes acting on xylan backbone (xylanase and β-xylosidase), and those acting on side chains (or accessory enzymes) notably α-arabinofuranosidase and acetyl-xylan esterase. The Asperigillus hemicellulases are characterized as having relatively low xylanase and β-xylosidase activities but high side chain removal activities. The activity ratio of side-chain acting enzymes to xylanase is higher than that of the Multifect enzyme, a commercial hemicellulase product. The potential of the novel hemicellulases in lignocelluloses bioprocessing was demonstrated with alkaline-pretreated switchgrass as lignocellulose substrate with hemicellulase supplemented with a ratio of xylanase activity to filter paper unit of 2:1. Supplement of Aspergillus hemicellulases to commercial ...
Lignocellulosic residues have been receiving growing interest as a promising source of polysaccharides, which can be converted into a variety of compounds, ranging from biofuels to bioplastics. Most of these can replace equivalent products traditionally originated from petroleum, hence representing an important environmental advantage. Lignocellulosic materials are theoretically unlimited, cheaper and may not compete with food crops. However, the conversion of these materials to simpler sugars usually requires cellulolytic enzymes. Being still associated with a high cost of production, cellulases are commonly considered as one of the main obstacles in the economic valorization of lignocellulosics. This work provides a brief overview of some of the most studied strategies that can allow an important reduction of cellulases consumption, hence improving the economy of lignocellulosics conversion. Cellulases recycling is initially discussed regarding the main processes to recover active enzymes and ...
Utilization of cellulases as substitute of chemical process gained huge momentum in the field of biotechnology Now there is dire need to find out un explore reveres of fungi possessing greater potential for efficient cellulase production. This boosted isolation of novel thermo tolerant fungal strains capable of producing the targeted product. In this investigation 70 thermophilic cellulolytic fungal strains were isolated. All the strains were screened via submerged fermentation. The strain showing highest CMCase activity was identified by conventional method i.e. based on morphology and microscopic features and confirmed by 18S rDNA gene sequencing, using specific ITS primers. The modified CTAB method was used for rapid extraction of DNA from thermo tolerant strain. The selected strain subsequently subjected to sequencing and phylogenetic analysis. The result indicates the selected strain was found to be T. dupontii. For strain improvement the T. dupontii was subjected to random mutagenesis by ...
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Several alkalophilic Bacillus spp. strains were selected for their capacity to produce alkaline cellulases. Culture supernatants of these strains showed optimal cellulase activities between pH 8 and 9 and they were stable from pH 6 to pH 12. A cellul
Summary of Facts and Submissions. I. The appeal lies from a decision of the Opposition Division revoking European patent 0 699 248, granted on European application No. 95 913 647.4.. II. The application as filed comprised 8 claims, reading, respectively, as follows:. 1. In a method for enhancing the feel and/or appearance and/or for providing color enhancement to a non-cotton containing cellulosic fabrics during manufacture of the fabric by treatment of the fabric with a composition comprising a naturally complete fungal cellulase composition which comprises exo-cellobiohydrolase type component(s) and endoglucanase type component(s) wherein the improvement comprises modifying the naturally complete fungal cellulase composition to comprise at least 10 weight percent of endoglucanase type components based on the total weight of protein in the fungal cellulase composition and be free of all CBH I type cellulase components.. 2. The method according to Claim 1 wherein said fungal cellulase ...
Non-cellulosomal processive endoglucanase 9I (Cel9I) from Clostridium thermocellum is a modular protein, consisting of a family-9 glycoside hydrolase (GH9) catalytic module and two family-3 carbohydrate-binding modules (CBM3c and CBM3b), separated by linker regions. GH9 does not show cellulase activity when expressed without CBM3c and CBM3b and the presence of the CBM3c was previously shown to be essential for endoglucanase activity. Physical reassociation of independently expressed GH9 and CBM3c modules (containing linker sequences) restored 60-70% of the intact Cel9I endocellulase activity. However, the mechanism responsible for recovery of activity remained unclear. In this work we independently expressed recombinant GH9 and CBM3c with and without their interconnecting linker in Escherichia coli. We crystallized and determined the molecular structure of the GH9/linker-CBM3c heterodimer at a resolution of 1.68 Å to understand the functional and structural importance of the mutual spatial orientation
Hyper-Productivity, Characterization, and Exploitation of a Cellulase Complex from a Novel Isolate of Aspergillus tubingenesis S2 using Lignocellulose-based Material
Effect of pH on the activity of (■) free cellulase, (●) immobilized cellulase, and (▲) immobilized cellulase + ionic liquid. Morphologies of immobilized c
Cellulase produced byTrichoderma viride acted on carboxymethyl cellulose with a Km of 4.9 g substrate per litre, showing a pH optimum at 4.5 and a temperature optimum at 55 °C. Ag+, Hg2+, Zn2+, Cu2+ and N3- were inhibitory.
The expression and distribution of digestive cellulases along the midgut of Cerambyx cerdo larvae were analyzed for the first time and are presented in this article. Four groups of larvae were examined: larvae developed in the wild; larvae taken from the wild and successively reared on an artificial diet based on polenta; and larvae hatched in the laboratory and reared on two different artificial diets. Seven endocellulase and seven β-D-glucosidase isoforms were detected in all midgut extracts of C. cerdo with a zymogram after native PAGE. We observed that C. cerdo larvae are capable of producing cellulase isoforms with different PAGE mobilities depending on the nutrient substrate. From our findings it can be assumed that, depending on the distribution of endocellulase and β-D-glucosidase, cellulose molecules are first fragmented in the anterior and middle midgut by endo-β-1,4-glucanase; subsequently, the obtained fragments are broken down by β-D-glucosidase mostly in middle midgut ...
glucosidases. A gene encoding endoglucanase, designated as cel12, was cloned from total RNA prepared from F. palustris grown at the expense of Avicel. The gene encoding Cel12 has an open reading frame of 732 bp, encoding a putative protein of 244 amino acid residues with a putative signal peptide residing at the first 18 amino acid residues of the N-terminus of the protein. Sequence analysis of Cel12 identified three consensus regions, which are highly conserved among fungal cellulases belonging to GH family 12. However, a cellulose-binding domain was not found in Cel12, like other GH family 12 fungal cellulases. Northern blot analysis showed a dramatic increase of cel12 mRNA levels in F. palustris cells cultivated on Avicel from the early to late stages of growth and the maintenance of a high level of expression in the late stage, suggesting that Cel12 takes a significant part in endoglucanase activity throughout the growth of F. palustris. Adventitious expression of cel12 in the yeast Pichia ...
Tobacco plants were used to produce a fungal cellulase, TrCel5A, via a transient expression system. The expression could be monitored...
The microbial degradation of cellulose and xylans requires several types of enzyme such as endoglucanases (EC 3.2.1.4), cellobiohydrolases (EC 3.2.1.91) (exoglucanases), or xylanases (EC 3.2.1.8) [(PUBMED:1886523)]. Structurally, cellulases and xylanases generally consist of a catalytic domain joined to a cellulose-binding domain (CBD) by a short linker sequence rich in proline and/or hydroxy-amino acids.. The CBD domain is found either at the N-terminal or at the C-terminal extremity of these enzymes. As it is shown in the following schematic representation, there are two conserved cysteines in this CBD domain - one at each extremity of the domain - which have been shown [(PUBMED:1761039)] to be involved in a disulphide bond. There are also four conserved tryptophan, two are involved in cellulose binding. The CBD of a number of bacterial cellulases has been shown to consist of about 105 amino acid residues [(PUBMED:1812490), (PUBMED:10973978)].. ...
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Members of family GH9 are mainly cellulases (EC 3.2.1.4), including primarily endo-glucanases and a few processive endo-glucanases. Indeed, as one of the first glycoside hydrolase families classified by hydrophobic cluster analysis, GH9 was previously known as Cellulase Family E [1, 2]. More recently, certain GH9 members from Clostridia [3] and Bacteroides [4, 5] have been shown to be endo-xyloglucanases (EC 3.2.1.151) or mixed-linkage endo-glucanases (EC 3.2.1.73). Exo-beta-glucosaminidases (EC 3.2.1.165) are also found in this family [6, 7]. All of the processive endoglucanases contain a family 3c CBM rigidly attached to the C-terminus of the GH9 catalytic domain (cd) [8]. This domain is part of the active site and is essential for processivity [8]. CBM3c domains bind weakly to cellulose as they lack several of the conserved aromatic residues that are important for cellulose binding in family 3a and family 3b members [9]. All known plant cellulases belong to GH9, and most of the other ...
Hemicellulase is used to clarify the juice of mandarin oranges. Although pectinase is used to clarify the juices of apples and grapes, the juice of mandarin oranges are clarified by the simultaneous use of pectinase and hemicellulase. In food processing, it is rare that the desired result can be obtained with the use of hemicellulase alone. Hemicellulases work in conjunction with cellulases and pectinases ...
Paper sludge is the largest solid waste stream produced by pulp and paper industry, and is also an attractive feedstock for emergent technologies based on processing of cellulosic biomass featuring enzymatic hydrolysis. This study focuses on investigating conversion of paper sludge to ethanol under industrially relevant conditions. A solids-fed simultaneous saccharification and fermentation laboratory reactor system capable of aseptic, semi-continuous metered delivery of paper sludge was developed to carry out experiments with hydrolysis mediated by commercial cellulase preparations and fermentation of glucose to ethanol mediated by Saccharomyces cerevisiae. Economically recoverable concentrations of ethanol were produced, and good material balance closure was achieved. Decreasing feeding frequency (feed additions per residence time) was found to allow the cellulase loading to be decreased at least two-fold with no decrease in cellulose conversion. Although decreasing feeding frequency results ...
Paper sludge is the largest solid waste stream produced by pulp and paper industry, and is also an attractive feedstock for emergent technologies based on processing of cellulosic biomass featuring enzymatic hydrolysis. This study focuses on investigating conversion of paper sludge to ethanol under industrially relevant conditions. A solids-fed simultaneous saccharification and fermentation laboratory reactor system capable of aseptic, semi-continuous metered delivery of paper sludge was developed to carry out experiments with hydrolysis mediated by commercial cellulase preparations and fermentation of glucose to ethanol mediated by Saccharomyces cerevisiae. Economically recoverable concentrations of ethanol were produced, and good material balance closure was achieved. Decreasing feeding frequency (feed additions per residence time) was found to allow the cellulase loading to be decreased at least two-fold with no decrease in cellulose conversion. Although decreasing feeding frequency results ...
The microbial degradation of cellulose and xylans requires several types of enzymes such as endoglucanases (EC 3.2.1.4), cellobiohydrolases (EC 3.2.1.91) (exoglucanases), or xylanases (EC 3.2.1.8) [1,2]. Fungi and bacteria produces a spectrum of cellulolytic enzymes (cellulases) and xylanases which, on the basis of sequence similarities, can be classified into families. One of these families is known as the cellulase family D [3] or as the glycosyl hydrolases family 8 [4,E1]. The enzymes which are currently known to belong to this family are listed below. ...
Books and Book Chapters. 1. Lactase production by Aspergillis Oryzae , Lambert publications, Germany, 2012. 2. Cellulase production by Aspergillus niger , Lambert publications , Germany, 2012. 3. Biological synthesis, Characterization and antimicrobial activity of silver nanoparticles from beetle leaves, In Nanoscience and drugdelivery. Apple science publication USA. Projects:. 1. Major Research project on Cellulase production by mutant fungal strain Aspergillus niger Sanctioned by University Grants Commision 2009. New Delhi, India. Abstracts. Indian Science Abstracts:. ...
The present invention provides a novel cellulase composition obtainable from Bacillus sp. CBS 669.93. A preferred cellulase has a calculated molecular weight of approximately 63 kD, a calculated isoelectric point of about 5 and a pH optimum on CMC of about 6 at 40 C. and 60 C.
Cellualse is one of the most important enzymes used in textile, detergent, paper, food and feed industries. Therefore, a study was undertaken to isolate Bacillus bacteria having the potential to produce cellulase from soil samples. 24 soil samples were analyzed and 54 presumptive Bacillus isolates were isolated after heating the soil samples at 80°C for 10 min. Among them 45 isolates showed enzyme activity ranging from 0.003 to 0.17 U/ml in test tubes containing 5 ml medium composed of (g/L) glucose 0.5 gm, peptone 0.75 gm, FeSO4 0.01 gm, KH2PO4 0.5 gm, and MgSO4 0.5 gm at 120 rpm, 37° C and pH 7. Among them 1RW, 2WS, 3YR, 4WT, 6 RR, and 9SS showed 0.17, 0.15, 0.14, 0.15, 0.147 and 0.14U/ml enzyme activities, respectively. Production of cellulase by these isolates was further scaled up to shake culture containing 50 ml medium similar to that used in test tube culture. Among the isolates 1 RW showed the maximum activity. This 1 RW was identified by API kit and showed that 59 % belongs to ...
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Omega Pharma Cellulase Anticellulite Mousse 150ml. Cellulase Crackling Mousse is a cellulite treatment that improves skin vasodilation by stimulating the microcirculation, improves penetration of active ingredients and provides a pleasant feeling of...
In support of this hypothesis , we observed a relatively high abundance of the larger antigen in the fruit after one day of propylene treatment, at which time synthesis of cellulase protein is in its early stages (Fig. 3, day 1). As ripening progresses, the lower molecular weight cellulase antigen became the predominant form (Fig . 3 , days 3 and 4), as would be expected if it were the final product of a post-translational processing pathway. The temporal expression of cellulase mRNA during propylene- induced ripening was investigated by northern analysis using two different probes. McGarvey Frank W. Percival , and Kristin R. A INTRODUCTION Softening of avocado fruit during autodigestion of the cell wall. a-D-galacturonide) ripening is mediated glycanohydrolase. C. C. 4], closely and cellulase [endo- are thought to be the principal enzymes involved in cell wall breakdown. , 1979; Awad and Young, 1979). , 1985; Christoffersen, 1987). Unripe fruit have a very low level of cellulase message which ...
Cellulase is often a plant enzyme that aids during the digestion of fibrous substances. Cellulase is utilized as a digestive aid and for the management of flatulence.Cellulase Cellulase is surely an enzyme which breaks down cellulose to beta-glucose. Human beings will not make cellulase of their bodies, and so are therefore struggling to use a lot … Read More. ...
Nucleic acid sequences encoding chimeric polypeptides that exhibit enhanced cellulase activities are disclosed herein. These nucleic acids may be expressed in hosts such as fungi, which in turn may be cultured to produce chimeric polypeptides. Also disclosed are chimeric polypeptides and their use in the degradation of cellulosic materials.
endo-β-1,4-glucanase / cellulase (EC 3.2.1.4); endo-β-1,4-xylanase (EC 3.2.1.8); β-glucosidase (EC 3.2.1.21); β-mannosidase (EC 3.2.1.25); β-glucosylceramidase (EC 3.2.1.45); glucan β-1,3-glucosidase (EC 3.2.1.58); licheninase (EC 3.2.1.73); exo-β-1,4-glucanase / cellodextrinase (EC 3.2.1.74); glucan endo-1,6-β-glucosidase (EC 3.2.1.75); mannan endo-β-1,4-mannosidase (EC 3.2.1.78); cellulose β-1,4-cellobiosidase (EC 3.2.1.91); steryl β-glucosidase (EC 3.2.1.104); endoglycoceramidase (EC 3.2.1.123); chitosanase (EC 3.2.1.132); β-primeverosidase (EC 3.2.1.149); xyloglucan-specific endo-β-1,4-glucanase (EC 3.2.1.151); endo-β-1,6-galactanase (EC 3.2.1.164); hesperidin 6-O-α-L-rhamnosyl-β-glucosidase (EC 3.2.1.168); β-1,3-mannanase (EC 3.2.1.-); arabinoxylan-specific endo-β-1,4-xylanase (EC 3.2.1.-); mannan transglycosylase (EC 2.4.1.- ...
endo-β-1,4-glucanase / cellulase (EC 3.2.1.4); endo-β-1,4-xylanase (EC 3.2.1.8); β-glucosidase (EC 3.2.1.21); β-mannosidase (EC 3.2.1.25); β-glucosylceramidase (EC 3.2.1.45); glucan β-1,3-glucosidase (EC 3.2.1.58); licheninase (EC 3.2.1.73); exo-β-1,4-glucanase / cellodextrinase (EC 3.2.1.74); glucan endo-1,6-β-glucosidase (EC 3.2.1.75); mannan endo-β-1,4-mannosidase (EC 3.2.1.78); cellulose β-1,4-cellobiosidase (EC 3.2.1.91); steryl β-glucosidase (EC 3.2.1.104); endoglycoceramidase (EC 3.2.1.123); chitosanase (EC 3.2.1.132); β-primeverosidase (EC 3.2.1.149); xyloglucan-specific endo-β-1,4-glucanase (EC 3.2.1.151); endo-β-1,6-galactanase (EC 3.2.1.164); hesperidin 6-O-α-L-rhamnosyl-β-glucosidase (EC 3.2.1.168); β-1,3-mannanase (EC 3.2.1.-); arabinoxylan-specific endo-β-1,4-xylanase (EC 3.2.1.-); mannan transglycosylase (EC 2.4.1.- ...
Cellulose is recalcitrant to deconstruction to glucose for use in fermentation strategies for biofuels and chemicals derived from lignocellulose. In Neurospora crassa, the transcriptional regulator, CLR-2, is required for cellulolytic gene expression
The filamentous fungus Trichoderma reesei (anamorph of Hypocrea jecorina) displays increased cellulase expression while growing on inducers such as lactose or cellulose. However, the mechanism of cellulase induction in T. reesei is not yet completely characterized. Here, a protein annotated as β-glucosidase (BGL3I) was found to be involved in cellulase induction in T. reesei. The effects of BGL3I on cellulase production have not yet been fully understood. Deletion of the bgl3i gene had no influence on the growth of T. reesei, but significantly increased its cellulase activities. Deletion of bgl3i also resulted in decreased extracellular galactosidase activity, but significantly increased transcription of lactose permeases, which might be involved in lactose transport. Furthermore, deletion of bgl3i enhanced the transcription levels of intracellular β-glucosidases cel1a, cel1b and the regulator xyr1, which are all essential for lactose induction in T. reesei. BGL3I was found to have a relatively high
The aim of this study was to enable the polymerase chain reaction (PCR) amplification of DNA fragments within endoglucanase gene(s) of Torula thermophila, by using degenerate primers so that the amplified fragment(s) could be used as homologous probe(s) for cloning of full-length endoglucanase gene(s). The design of the degenerate PCR primers was mainly based on the endoglucanase sequences of other fungi. The endoglucanase gene sequence of Humicola insolens was the only sequence from a thermophilic fungus publicly available in the literature. Therefore, the endoglucanase sequences of the two Trichoderma species, Trichoderma reesei and Trichoderma longibrachiatum, were used to generalize the primers. PCR amplification of T. thermophila genomic DNA with these primers resulted in a specific amplification. The specificity of the amplified fragment was shown by Southern hybridization analysis using egl3 gene of T. reesei as probe. This result suggested that the degenerate primers used in this study ...
An analysis of the recently published genome sequence of Cytophagahutchinsonii revealed an unusual collection of genes for an organism that can attackcrystalline cellulose. Consequently, questions were being raised by cellulase scientists, as towhat mechanism this organism uses to degrade its insoluble substrates. Cellulose, being ahighly polymeric compound and insoluble in water, cannot enter the cell walls ofmicroorganisms. Cellulose-degrading enzymes have therefore to be located on the surface ofthe cell wall or released extracellularly. The location of most cellulase enzymes has beenstudied. However, basic information on C. hutchinsonii cellulases is almost non-existent. Inthe present study, the location, formation and biosynthetic regulation of cellulases in C.hutchinsonii were demonstrated on different substrates. Various fractions isolated from C.hutchinsonii after cell rupture were assayed for carboxymethyl-cellulase activity (CMC).The cellulases were found to be predominantly cell-free during
Trichoderma reesei is a key cellulase source for economically saccharifying cellulosic biomass for the production of biofuels. Lignocellulose hydrolysis at temperatures above the optimum temperature of T. reesei cellulases (~50°C) could provide many significant advantages, including reduced viscosity at high-solids loadings, lower risk of microbial contamination during saccharification, greater compatibility with high-temperature biomass pretreatment, and faster rates of hydrolysis. These potential advantages motivate efforts to engineer T. reesei cellulases that can hydrolyze lignocellulose at temperatures ranging from 60-70°C. A B-factor guided approach for improving thermostability was used to engineer variants of endoglucanase I (Cel7B) from T. reesei (TrEGI) that are able to hydrolyze cellulosic substrates more rapidly than the recombinant wild-type TrEGI at temperatures ranging from 50-70°C. When expressed in T. reesei, TrEGI variant G230A/D113S/D115T (G230A/D113S/D115T Tr
Cellulosic ethanol produced by microbial fermentation from feedstocks has been proposed to replace fossil fuels in transportation. A key step in cellulosic ethanol production is to break down cellulose into glucose and hemicellulose into xylose, which can subsequently be converted into ethanol by fermentative microbes. Therefore, finding efficient cellulases is important to bioethanol production, as well as for hydrolyzing feedstocks into sugars in general. Neocallimastix species is one of the major anaerobic fungi in the rumen of water buffalo capable of efficiently digesting cellulosic biomass [1-4]. Such anaerobic fungi are potential sources for highly active cellulolytic enzymes that are useful for cellulose hydrolysis [5-7]. Plant cell wall degrading enzymes from rumen fungi such as Neocallimastix patriciarum may be used for the production of industrial materials from plant biomass. These enzymes may also improve the fiber properties of cotton for manufacturing or clothes. The simple sugars ...
Cellulase (endo-1,4-β-D-glucanase) (Bacillus amyloliquefaciens) [ME-CELBA] - High purity recombinant Cellulase (endo-1,4-β-D-glucanase) (Bacillus amyloliquefaciens) for use in research, biochemical enzyme assays and in vitro diagnostic analysis. EC 3.2.1.4 CAZy Family: GH5 Recombinant. From Bacillus amyloliquefaciens. In 3.2 M ammonium sulphate. Specific activity: ~ 80 U/mg (40oC, pH 6.0 on CM-cellulose 4M); ~ 160 U/mg (60oC, pH 6.0 on CM-cellulose 4M). Stability: | 2 years at 4oC.
Cellulolytic enzymes capable of efficiently degrading crystalline cellulose are a complex mixture of endo- (endoglucanases) and exo-acting (cellobiohydrolases) enzymes. One approach to separating these enzymes is affinity chromatography. A new ligand, p-aminophenyl l-thio-β-D-cellobioside (APTC), is introduced for this purpose. The property of APTC in affinity chromatography is demonstrated using Trichoderma reesei cellulases. The behavior of these enzymes on APTC-affinity column was essentially equivalent to that reported for the same enzymes on p-aminobenzyl 1-thio-β- D-cellobioside (ABTC)-columns; ABTC being the traditional ligand for affinity chromatography of exocellulases. The primary advantage of the APTC ligand is its ease of preparation. The affinity between CBHs and APTC may be considerably affected by nonspecific interactions. In this study, the significance of nonspecific protein/matrix interactions in affinity chromatography of cellulolytic enzymes is evaluated. The role of pH, ...
Effect of Natural and Pretreated Soybean Hulls on Enzyme Production by Trichoderma reesei. (A. M. Coffman, Q. Li, L.-K. Ju) Journal of the American Oil Chemists Society 91 (8), 1331-1338 (2014). View Article. Promoting Pellet Growth of Trichoderma reesei Rut C30 by Surfactants for Easy Separation and Enhanced Cellulase Production. (N.V. Callow and L.-K. Ju) Enzyme and Microbial Technology 50(6-7), 311-317 (2012). View Article. Cellulase production by continuous culture of Trichoderma reesei Rut C-30 using acid hydrolysate prepared to retain more oligosaccharides for induction. (C.-M. Lo, Q. Zhang, N. V. Callow and L.-K. Ju) Bioresource Technology. 101(2), 717-23 (2010). View Article. Cell immobilization with polyurethane foam for retaining Trichoderma reesei cells during foam fractionation for cellulase collection. (Q. Zhang, C.-M. Lo, and L.-K. Ju) Applied Biochemistry and Biotechnology 156, 12-23 (2009). View Article. Cellulase production by cocultures of Hypocrea jecorina Rut C30 and Candida ...
phdthesis{a42f4c58-3e50-41b9-87ac-a3c37465f8bc, abstract = {The enzymatic degradation of wood polysaccharides such as cellulose and hemicellulose is an important process in nature. In addition, cellulases and hemicellulases can be used in industrial applications. Fuel ethanol can potentially be produced from wood by enzymatic hydrolysis of cellulose followed by yeast fermentation of the formed sugars.,br/,,br, ,br/,,br, In this thesis, fungal glycoside hydrolases, cellulases and hemicellulases were studied with the aim of increasing our knowledge of the mechanisms involved in the enzymatic hydrolysis of cellulose and lignocellulose. The focus was mainly on cellulases from the filamentous fungus Trichoderma reesei. However, lignocellulose also contains hemicellulose and studies of hemicellulases are included,br/,,br, ,br/,,br, In Paper I-IV the mechanisms involved in cellulose degradation were investigated. Features of enzymatic cellulose hydrolysis such as synergism, decreasing hydrolysis rate ...
The interaction between cellulase enzymes and their substrates is of central importance to several technological and scientific challenges. Here we report that the binding of cellulose binding modules (CBM) from Trichoderma reesei cellulases Cel6A and Cel7A show a major difference in how they interact with substrates originating from wood compared to bacterial cellulose. We found that the CBM from TrCel7A recognizes the two substrates differently and as a consequence shows an unexpected way of binding. We show that the substrate has a large impact on the exchange rate of the studied CBM, and moreover, CBM-TrCel7A seems to have an additional mode of binding on wood derived cellulose but not on cellulose originating from bacterial source. This mode is not seen in double CBM (DCBM) constructs comprising both CBM-TrCel7A and CBM-TrCel6A. The linker length of DCBMs affects the binding properties, and slows down the exchange rates of the proteins and thus, can be used to analyze the differences ...
TY - JOUR. T1 - Adsorption of monocomponent enzymes in enzyme mixture analyzed quantitatively during hydrolysis of lignocellulose substrates. AU - Varnai, Anikó. AU - Viikari, Liisa. AU - Marjamaa, Kaisa. AU - Siika-aho, Matti. PY - 2010. Y1 - 2010. N2 - The adsorption of purified Trichoderma reesei cellulases (TrCel7A, TrCel6A and TrCel5A) and xylanase TrXyn11 and Aspergillus niger β-glucosidase AnCel3A was studied in enzyme mixture during hydrolysis of two pretreated lignocellulosic materials, steam pretreated and catalytically delignified spruce, along with microcrystalline cellulose (Avicel). The enzyme mixture was compiled to resemble the composition of commercial cellulase preparations. The hydrolysis was carried out at 35 °C to mimic the temperature of the simultaneous saccharification and fermentation (SSF). Enzyme adsorption was followed by analyzing the activity and the protein amount of the individual free enzymes in the hydrolysis supernatant. Most enzymes adsorbed quickly at ...
TY - JOUR. T1 - Stability analysis of Bacillus stearothermopilus L1 lipase fused with a cellulose-binding domain. AU - Hwang, Sangpill. AU - Ahn, Ik Sung. N1 - Funding Information: ^ÅâåçïäÉÇÖÉãÉåí This work was made possible with funding provided by the Korea Science & Engineering Foundation to Advanced Environmental Biotechnology Research Center at POSTECH.. PY - 2005. Y1 - 2005. N2 - This study was designed to investigate the stability of a lipase fused with a cellulose-binding domain (CBD) to cellulase. The fusion protein was derived from a gene cluster of a CBD fragment of a cellulase gene in Trichoderma hazianum and a lipase gene in Bacillus stearothermophilus L1. Due to the CBD, this lipase can be immobilized to a cellulose material. Factors affecting the lipase stability were divided into the reaction-independent factors (RIF), and the reaction-dependent factors (RDF). RIF includes the reaction conditions such as pH and temperature, whereas substrate limitation and product ...
TY - JOUR. T1 - Characterization of a multidomain cellulase from an extremely thermophilic anaerobe strain NA10. AU - Miyake, Katsuhide. AU - Machida, Yuichi. AU - Hattori, Kouji. AU - Iijima, Shinji. N1 - Copyright: Copyright 2017 Elsevier B.V., All rights reserved.. PY - 1998. Y1 - 1998. N2 - The nucleotide sequence of a β-glucanase gene from an extremely thermophilic anaerobe NA10 was determined. The open reading frame extended over 3000 bp and encoded a polypeptide with a molecular mass of 113 kDa. The deduced amino acid sequence of this protein exhibited high homology to a bifunctional cellulase CelB of Caldocellum saccharolyticum. Based on the homology to GelB, the NA10 β-glucanase appears to comprise three domains: N- terminal, central, and C-terminal domains. Among these, N- and C-terminal domains apper to be catalytic domains, and the central domain to be a cellulose binding domain. These domains were joined with each other by proline and threonine rich segments (PT box). The ...
Various enzymatic cocktails were produced from two Trichoderma reesei strains, a cellulase hyperproducer strain and a strain with β-glucosidase activity overexpression. By using various carbon sources (lactose, glucose, xylose, hemicellulosic hydrolysate) for strains growth, contrasted enzymatic activities were obtained. The enzymatic cocktails presented various levels of efficiency for the hydrolysis of cellulose Avicel into glucose, in presence of xylans, or not. These latter were also hydrolyzed with different extents according to cocktails. The most efficient cocktails (TR1 and TR3) on Avicel were richer in filter paper activity (FPU) and presented a low ratio FPU/β-glucosidase activity. Cocktails TR2 and TR5 which were produced on the higher amount of hemicellulosic hydrolysate, possess both high xylanase and β-xylosidase activities, and were the most efficient for xylans hydrolysis. When hydrolysis of Avicel was conducted in presence of xylans, a decrease of glucose release occurred for all
The commonly used laboratory bacterium Escherichia coli normally does not produce and secrete cellulases due to its complex bilayer membrane structure and poor secretory apparatus. In our previous study, the cellulolytic E. coli strain ZH-4 with extracellular cellulase activity was found in the bovine rumen. In this study, we demonstrate that the secretion of cellulase is a common feature of E. coli isolates from the rumen of animals such as sheep and cattle. Physiological phenotype characterization of these E. coli isolates, together with genome, transcriptome, and comparative genomics analysis, suggests their adaption to the rumen niche. The higher growth rate of the isolated strains under aerobic conditions meets the competitive requirements of the strains in rumen microecosystem, while anaerobic accumulation of reduced H2 and succinate is hypothesized to be the results of adaptation to the rumen environment. Cellulase secretion increased significantly when the molecular chaperone genes ibpA ...
The enzyme diversity from the cellulolytic system produced by grown on crystalline cellulose like a sole carbon and energy source was explored by two-dimensional electrophoresis. proteins outlined in the NCBI database. Using Trap-Dock PCR and DNA walking, seven genes encoding new dockerin-containing proteins were cloned and sequenced. Some of these genes are clustered. Enzymes encoded by these genes belong to glycoside hydrolase family members GH2, GH9, GH10, GH26, GH27, and GH59. Except for members of family GH9, which consists of only cellulases, the new modular glycoside hydrolases found out in this work could be involved in the degradation of different hemicellulosic substrates, such as xylan or galactomannan. Cellulose, a long polymer of -1,4-glucose, is the major component of the herb cell wall (39). Cellulolytic bacteria and fungi secrete many different types of cellulases to catalyze efficient degradation of this recalcitrant substrate. Many cellulolytic, anaerobic microorganisms secrete ...
Plasma-assisted pretreated wheat straw was investigated for cellulase and xylanase production by Trichoderma reesei fermentation. Fermentations were conducted with media containing washed and unwashed plasma-assisted pretreated wheat straw as carbon source which was sterilized by autoclavation. To account for any effects of autoclavation, a comparison was made with unsterilized media containing antibiotics. It was found that unsterilized washed plasma-assisted pretreated wheat straw (which contained antibiotics) was best suited for the production of xylanases (110 IU ml(-1)) and cellulases (0.5 filter paper units (FPU) ml(-1)). Addition of Avicel boosted enzyme titers with the highest cellulase titers (1.5 FPU ml(-1)) found with addition of 50 % w/w Avicel and with the highest xylanase production (350 IU ml(-1)) reached in the presence of 10 % w/w Avicel. Comparison with enzyme titers from other nonrefined feedstocks suggests that plasma pretreated wheat straw is a promising and suitable ...
Cellulase is any of several enzymes produced chiefly by fungi, bacteria, and protozoans that catalyze cellulolysis, the decomposition of cellulose and of some related polysaccharides. The name is also used for any naturally occurring mixture or complex of various such enzymes, that act serially or synergistically to decompose cellulosic material. Cellulases break down the cellulose molecule into monosaccharides (simple sugars) such as beta-glucose, or shorter polysaccharides and oligosaccharides. Cellulose breakdown is of considerable economic importance, because it makes a major constituent of plants available for consumption and use in chemical reactions. The specific reaction involved is the hydrolysis of the 1,4-beta-D-glycosidic linkages in cellulose, hemicellulose, lichenin, and cereal beta-D-glucans. Because cellulose molecules bind strongly to each other, cellulolysis is relatively difficult compared to the breakdown of other polysaccharides such as starch.[2] Most mammals have only ...
Method of preparation and some properties of amorphous cellulose nanoparticles (ANP) have been described in this paper. It was shown that ANP have spherical shape and are characterized by high degree of pantamorphia, low DP and increased content of sulfonic groups. The amorphous nanoparticles of cellulose are completely hydrolyzed by cellulolytic enzymes with forming of glucose. Concentrated paste of ANP has expressed thickening properties and therefore its additive can prevent phase separation of water dispersions of various substances. Low-acidic and soft nanoparticles can be used in cosmetic formulation for gentle skin peeling. Moreover, due to increased content of acidic functional groups, ANP can immobilize various therapeutically-active substances (TAS) containing basic functional groups. The ANP-TAS complexes can be used in remedies aimed for effective care and cure of the skin.
Cellulase is any of several enzymes produced chiefly by fungi, bacteria, and protozoans that catalyze cellulolysis, the decomposition of cellulose and of some related pol
Background The in-depth understanding of the enzymatic hydrolysis of cellulose with heterogeneous morphology (that is, crystalline versus amorphous) may help develop better cellulase cocktail mixtures and biomass pretreatment, wherein cost-effective release of soluble sugars from solid cellulosic materials remains the largest obstacle to the economic viability of second generation biorefineries. Results In addition to the previously developed non-hydrolytic fusion protein, GC3, containing a green fluorescent protein (GFP) and a family 3 carbohydrate-binding module (CBM3) that can bind both surfaces of amorphous and crystalline celluloses, we developed a new protein probe, CC17, which contained a mono-cherry fluorescent protein (CFP) and a family 17 carbohydrate-binding module (CBM17) that can bind only amorphous cellulose surfaces. Via these two probes, the surface accessibilities of amorphous and crystalline celluloses were determined quantitatively. Our results for the enzymatic hydrolysis of ...
Hydrolysis of cellulose to glucose is the most critical step in bioconversion of lignocellulosic biomass to fuels and chemicals. Cellulases and homogenous acids are widely used for cellulose hydrolysis. However, cellulases can only be used at moderate conditions and need longer time to achieve satisfactory cellulose hydrolysis. Homogenous acids can tolerate higher temperature, but they have limitations and issues such as equipment corrosion, recycling and wastewater treatment. To address these issues, heterogeneous solid acids have recently drawn a lot of attention for hydrolyzing cellulose. Traditional solid acids such as sulfonated carbon and resins, however, are not as effective as homogenous acids and cellulases in hydrolyzing cellulose because they have poor access/affinity to cellulose. In this study, a series of porous polymeric solid acids were synthesized for cellulose hydrolysis. These cellulase-mimetic solid acids have hydroxyl, halide, or boronic acid as cellulose-binding group in ...
Disclosed are variants of Humicola grisea CeI7A (CBH1.1), H. jecorina CBH1 variant or S. thermophilium CBH1, nucleic acids encoding the same and methods for producing the same. The variant cellulases have the amino acid sequence of a glycosyl hydrolase of family 7A wherein one or more amino acid residues are substituted.
Abdulrahman AO, Huisingh D (2018). The role of biomass as a cleaner energy source in Egypts energy mix. Journal of Cleaner Production 172:3918-3930. Ahmed S, Bashir A, Saleem H, Mubshara S, Jamil A (2009). Production and purification of cellulose - degrading enzymes from a filamentous fungus. Pakistan Journal of Botany 41(3):1411-1419. Bakker RRC, Elbersen HW, Poppens RP, Lesschen JP (2013). Rice straw and wheat straw. Potential feedstocks for the biobased economy. NL Agency. Report No. 448025. Retrieved from https://library.wur.nl/WebQuery/wurpubs/448025. Bayer EA, Morag E, Lamed R (1991). The cellulosome - a treasure- trove for biotechnology. Journal of Biological Chemistry 266(253):9241-9250. Béguin P, Aubert J-P (1994). The biological degradation of cellulose. Federation of European Microbiological Societies Microbiology Reviews 13(1):25-58. Benoit I, Culleton H, Zhou M, Difalco M, Osorio GA, Battaglia E, … Vries R P (2015). Closely related fungi employ diverse enzymatic strategies to ...
Cellobiohydrolases (CBHs), belonging to glycoside hydrolase families 6 and 7 (GH6 and GH7), are the major components of cellulase systems of filamentous fungi involved in biodegradation of cellulose in nature. Previous studies demonstrated that N-linked glycans in the catalytic domains of GH7 CBHs significantly affect the enzyme activity against cellulosic substrates. The influence of N-linked glycans on the activity and processivity of recombinant GH6 CBH II from Penicillium verruculosum (PvCel6A) was studied using site-directed mutagenesis of the respective Asn residues. Depending on the position of N-glycans on the surface of a protein globule, they affected the enzyme activity against cellulose either negatively or positively. The decrease or increase in the degree of processivity of recombinant forms of PvCel6A generally correlated with activity changes against Avicel. The mechanism of the N-glycan influence seems to be universal for GH6 and GH7 CBHs. The observed effects for CBHs from both ...
Trichoderma reesei is a key fungus for industrial production of lignocellulolytic enzymes. The genome sequences of the T. reesei hyper-cellulolytic strain RUT-C30 and its parental strain QM6a were compared at the nucleotide level. Approximately 97% of the 87 genomic-sequence scaffolds of T. reesei QM6a (33Mb) were found to have the corresponding nucleotide in the 182 genome-sequence scaffolds of RUT-C30 (32Mb). There are 455 loci within the QM6 sequence not detected in the RUT-C30 sequence. Regions at the termini of QM6a scaffolds as well as 14 small scaffolds do not have corresponding regions in RUT-C30 genomic scaffolds. Seventy-eight protein-encoding genes are included within these regions. Mutated nucleotide(s) in 2,371 positions, including short insertion/deletions (indels), were detected in the aligned regions. The predicted protein-coding regions of 97 gene models contain mutations, 34 of which were not previously described. Twenty-seven out of 34 newly discovered genes were found to have ...
The industrial applications of cellulases are mostly limited by the costs associated with their production. Optimized production pathways are therefore desirable. Based on their enzyme inducing capacity, celluloses are commonly used in fermentation media. However, the influence of their physiochemical characteristics on the production process is not well understood. In this study, we examined how physical, structural and chemical properties of celluloses influence cellulase and hemicellulase production in an industrially-optimized and a non-engineered filamentous fungus: Trichoderma reesei RUT-C30 and Neurospora crassa. The performance was evaluated by quantifying gene induction, protein secretion and enzymatic activities. Among the three investigated substrates, the powdered cellulose was found to be the most impure, and the residual hemicellulosic content was efficiently perceived by the fungi. It was furthermore found to be the least crystalline substrate and consequently was the most readily
The 2018 Gordon Research Seminar on Cellulosomes, Cellulases and Other Carbohydrate Modifying Enzymes (GRS) will be held in Andover, NH. Apply today to reserve your spot.
Meruliporia incrassata ATCC ® 11236™ Designation: Madison 563 Application: fungus resistance testing produces endoglucanase Cel 25 produces endoglucanase Cel 49 produces endoglucanase Cel 57 testing wood preservatives
One.Product presentation Cellulase is extracted from the fermentation solution which is produced by fermenting Trichoderma koningii which is a fruitful strain. This enzyme is being used in textile,feed,alcohol,fuel alcohol,beer fermentation,extract medicine,etc. Two.Product specification and quality index Item International Trade Leads - Importers And Exporters. INTERNATIONAL TRADE DIRECTORY for b2b trade. Suppliers and business opportunities in USA, EUROPE, ASIA and around the world..
p>The checksum is a form of redundancy check that is calculated from the sequence. It is useful for tracking sequence updates.,/p> ,p>It should be noted that while, in theory, two different sequences could have the same checksum value, the likelihood that this would happen is extremely low.,/p> ,p>However UniProtKB may contain entries with identical sequences in case of multiple genes (paralogs).,/p> ,p>The checksum is computed as the sequence 64-bit Cyclic Redundancy Check value (CRC64) using the generator polynomial: x,sup>64,/sup> + x,sup>4,/sup> + x,sup>3,/sup> + x + 1. The algorithm is described in the ISO 3309 standard. ,/p> ,p class=publication>Press W.H., Flannery B.P., Teukolsky S.A. and Vetterling W.T.,br /> ,strong>Cyclic redundancy and other checksums,/strong>,br /> ,a href=http://www.nrbook.com/b/bookcpdf.php>Numerical recipes in C 2nd ed., pp896-902, Cambridge University Press (1993),/a>),/p> Checksum:i ...
1G9J: Structures of mutants of cellulase Cel48F of Clostridium cellulolyticum in complex with long hemithiocellooligosaccharides give rise to a new view of the substrate pathway during processive action
TY - GEN. T1 - Trichoderma cellulases. T2 - 1st European Conference on Fungal Genetics. AU - Penttilä, Merja. AU - Ilmen, Marja. AU - Keränen, Sirkka. AU - Nakari, Tiina. AU - Nyyssönen, Eini. AU - Onnela, Maija-Leena. AU - Saloheimo, Anu. N1 - Project code: BIO8004. PY - 1992. Y1 - 1992. M3 - Conference article in proceedings. BT - 1st European Conference on Fungal Genetics. PB - University of Nottingham. CY - Nottingham. Y2 - 20 August 1992 through 23 August 1992. ER - ...
TY - JOUR. T1 - The antimicrobial reagent role on the degradation of model cellulose film. AU - Jausovec, D.. AU - Angelescu, Daniel. AU - Voncina, B.. AU - Nylander, Tommy. AU - Lindman, Björn. PY - 2008. Y1 - 2008. N2 - The effect of the antimicrobial agent TMPAC (3-(trimethoxysilyl)-propyldimethyloctadecyl ammonium chloride) on the cellulase activity oil model cellulose substrate was investigated by in situ-null ellipsometry. The cellulases used were extracted from Trichoderma virlde and Aspergillus niger, and the model cellulose film was prepared by spin-coating silicon oxide wafers with cellulose solubilized in N-methylmorpholine-N-oxide/dimethyl sulfoxide solution. Upon enzyme addition to the previously equilibrated cellulose film, the initial enzyme adsorption oil the substrate was followed by an overall decrease in film mass owing to enzymatic digestion of the cellulose. The loss of cellulose film mass was associated with a non-monotonously behavior of the cellulose film thickness. The ...
Hypocrea jecorina (anamorph Trichoderma reesei) is a filamentous ascomycete of industrial importance due to its hydrolases (e.g., xylanases and cellulases). The regulation of gene expression can influence the composition of the hydrolase cocktail, and thus, transcription factors are a major target of current research. Here, we design an approach for identifying a repressor of a xylanase-encoding gene. We used streptavidin affinity chromatography to isolate the Xylanase promoter-binding protein 1 (Xpp1). The optimal conditions and templates for the chromatography step were chosen according to the results of an electrophoretic mobility shift assay performed under repressing conditions, which yielded a DNA-protein complex specific to the AGAA-box (the previously identified, tetranucleotide cis-acting element). After isolating AGAA-box binding proteins, the eluted proteins were identified with Nano-HPLC/tandem MS-coupled detection. We compared the identified peptides to sequences in the H. jecorina genome
This project is aimed at using large-scale, high-performance computing to gain insights into the primary routes that nature uses to degrade plant cell walls. The overall goal is to enable rational design of superior biological catalysts for conversion of biomass to sugars for renewable liquid fuels. Project researchers will focus on the prevalent biological route to cellulose conversion: enzymatic hydrolysis by cellulases.
Some exocellulases, most of which belong to the glycoside hydrolase family 48 (GH48, formerly known as cellulase family L), act at the reducing ends of cellulose and similar substrates. The CelS enzyme from Clostridium thermocellum is the most abundant subunit of the cellulosome formed by the organism. It liberates cellobiose units from the reducing end by hydrolysis of the glycosidic bond, employing an inverting reaction mechanism [2]. Different from EC 3.2.1.91, which attacks cellulose from the non-reducing end ...
I was thinking of trying something about candida again and searched vitacost for anti-fungals and cleanses. Among the usual combos with fiber, probiotics and anti-fungals like garlic and herbs, there were some that contained enzymes as well or instead.http://www.vitacost.com/NSI-Candida-Yeast-Man...
The crystal structure of Cel48F, a cellulosome component of C. cellulolyticum, revealed the active center at the junction of the cleft and tunnel regions, where Glu55 is the proposed proton donor in the cleavage reaction, and the corresponding base was initially proposed to be either Glu44 or Asp230 [8]. The structure of the catalytic module of Cel48S of C. thermocellum showed a similar tunnel-shaped substrate-binding region formed by the alpha helices in the protein. The hydrolysis of the cellulose chain in Cel48S appeared to involve Glu87 (the equivalent of Glu55 in C. cellulolyticum Cel48F) as an acid to protonate the glycosidic oxygen atom and Tyr351 as a base to extract a proton from the nucleophilic water molecule that attacks the anomeric carbon atom. More recent studies of Cel48F failed to unambiguously identity the catalytic base in the cleavage reaction [7]. A recent experimental study in Thermobifida fusca Cel48A confirmed that aspartic acid (Asp225) is the catalytic base in family 48 ...
15112DNAMyceliophthora thermophila 1ctttccagca ca 1228DNAMyceliophthora thermophila 2gaaaggtc 831188DNAMyceliophthora thermophila 3cgacttgaaa cgccccaaat gaagtcctcc atcctcgcca gcgtcttcgc cacgggcgcc 60gtggctcaaa gtggtccgtg gcagcaatgt ggtggcatcg gatggcaagg atcgaccgac 120tgtgtgtcgg gctaccactg cgtctaccag aacgattggt acagccagtg cgtgcctggc 180gcggcgtcga caacgctgca gacatcgacc acgtccaggc ccaccgccac cagcaccgcc 240cctccgtcgt ccaccacctc gcctagcaag ggcaagctga agtggctcgg cagcaacgag 300tcgggcgccg agttcgggga gggcaattac cccggcctct ggggcaagca cttcatcttc 360ccgtcgactt cggcgattca gacgctcatc aatgatggat acaacatctt ccggatcgac 420ttctcgatgg agcgtctggt gcccaaccag ttgacgtcgt ccttcgacca gggttacctc 480cgcaacctga ccgaggtggt caacttcgtg acgaacgcgg gcaagtacgc cgtcctggac 540ccgcacaact acggccggta ctacggcaac atcatcacgg acacgaacgc gttccggacc 600ttctggacca acctggccaa gcagttcgcc tccaactcgc tcgtcatctt cgacaccaac 660aacgagtaca acacgatgga ccagaccctg gtgctcaacc tcaaccaggc cgccatcgac 720ggcatccggg ccgccggcgc gacctcgcag tacatcttcg ...
article{569d6702-1354-4b3b-8b97-3c11c9807854, abstract = {Metabolic stress is a phenomenon often discussed in conjunction with recombinant protein production in Escherichia coli. This investigation shows how heterologous protein production, and the presence of host cell proteases is related to: i) IPTG induction, ii) cell mass concentration at the time of induction and iii) the presence of metabolites (glutamic acid or those from TSB) during the post induction phase of high-cell-density (HCD) fed-batch cultivations. Two thermostable xylanase variants and one thermostable cellulase, all originating from Rhodothermus marinus were expressed in E. coli strain BL21 (DE3). A three-fold difference in the specific activity of both xylanase variants [between 7000 and 21000 U / (g cdw)], was observed under the different conditions tested. Upon induction at high cell-mass concentrations employing a nutrient feed devoid of the metabolites above, the specific activity of the xylanase variants, was initially ...
We have used chemostat cultivations at specific growth rates and cell densities to characterise the transcriptome and proteome of T.reesei in order to understand the molecular bases of low growth protein production phenotype. The stability of the cultivations was monitored with online and off-line measurements, including a monitoring for stability of transcription of the 31 reporter genes covering essential cellular processes [66].. We used the strain Rut-C30, instead of the sequenced QM6a strain, due to its improved protein production capabilities. The mutations in Rut-C30 in comparison to QM6a have been described genome wide [67, 68] and the phenotype of three of them have been studied. For the single major multi gene deletion of Rut-C30 in scaffold 15, it has been shown that it has no impact to cellulase production on lactose containing medium [68]. The glucosidase II alpha subunit frameshift of Rut-C30 improves protein production by changing the glycosylation pattern of secreted proteins ...
The ability of termites to eat wood and break the cellulose down into glucose quickly and efficiently have made studying the insects a point of focus for a number of scientists, including Nobel laureate Steven Chu, now Director of the Lawrence Berkeley Labs, who would like to apply that capability...
Cellulase is utilized for industrial food stuff processing in espresso. It performs hydrolysis of cellulose in the course of drying of beans. Cellulase is Utilized in the fermentation of biomass into biofuels, Whilst this method is comparatively experimental At the moment. Cellulase is employed to handle click here Phytobezoars, a sort of cellulose bezoar located in the human abdomen ...
Enzymes have a very wide range of functions in living organisms. Both signal transduction and regulation of cellular activity rely on enzymes, especially kinases and phosphatases. Enzymes also involve in movement by catalyzing the hydrolysis of ATP on myosin to make muscle contractions and can act as part of the cytoskeleton involved in transporting intracellular substances. ATP enzyme in the cell membrane as the ion pump involves in active transport.. In this articles, there introduces several enzymes biological functions from Creative Enzymes which may greatly help your study researches.. Cellulase. Since the study of cellulase has entered the molecular level, people have gradually explored the structure and function, gene regulation and genetic modification of cellulase.. Chitinase. Chitinase plays a very important physiological role in various organisms. In recent years, a variety of chitinases and chitinase-like proteins have been found in mammals, which plays a very important role in the ...