The genes encoding the three subunits of the primary ABC transporter Ota of the methanogenic archaeon Methanosarcina mazei Gö1 were cloned in an expression vector (pBAD24) and transformed into the glycine betaine transport-negative mutant Escherichia coli MKH13. Ota was produced as demonstrated by Western blotting. Uptake studies revealed that Ota catalyzed the transport of glycine betaine in E. coli MKH13(pBAD-Ota) with a Km of 10±5 μM and a maximal velocity of 1.5±0.5 nmol min⁻¹ mg protein⁻¹. Transport was ATP dependent. Ota was activated by salinity gradients, but only marginally by sugar gradients across the membrane. Glycine betaine transport was inhibited to a small extent by an excess of dimethylglycin or proline betaine, but not by sarcosine or glycine ...

Acetate is the major source of biological methane in both freshwater and marine environments (1, 2). Only two genera (Methanosarcina and Methanosaeta) of acetate-utilizing methane-producing microbes are known, of which Methanosarcina species have been researched to a greater extent. Most investigations have focused on Methanosarcina barkeri and Methanosarcina mazei, for which electron transport in the pathway of acetate conversion to methane is dependent on the production and consumption of H2, although the majority of Methanosarcina species are unable to metabolize H2 (3). On the other hand, all Methanosarcina species investigated transfer the methyl group of acetate to methane similarly, beginning with the CO dehydrogenase/acetyl coenzyme A (CoA) complex (Cdh), which cleaves activated acetate into methyl and carbonyl groups (4). The Cdh transfers the methyl group to tetrahydrosarcinapterin (THSPT), followed by transfer from CH3-THSPT to coenzyme M (HS-CoM), a reaction catalyzed by a membrane ...

Methanosarcina mazei ATCC ® BAA-159D-5™ Designation: Genomic DNA from Methanosarcina mazei strain DSM 3647 TypeStrain=False Application:

The production of methane by methanogens is dependent on numerous iron-sulfur (Fe-S) cluster proteins; yet, the machinery involved in Fe-S cluster biogenesis in methanogens remains largely unknown. Methanogen genomes encode uncharacterized homologs of the core components of the ISC (IscS and IscU) and SUF (SufBC) Fe-S cluster biogenesis systems found in bacteria and eukaryotes. Methanosarcina acetivorans contains three iscSU and two sufCB gene clusters. Here, we report genetic and biochemical characterization of M. acetivorans iscSU2. Purified IscS2 exhibited pyridoxal 5′- phosphate-dependent release of sulfur from L-cysteine. Incubation of purified IscU2 with IscS2, cysteine, and iron (Fe2+) resulted in the formation of [4Fe-4S] clusters in IscU2. IscU2 transferred a [4Fe-4S] cluster to purified M. acetivorans apo-aconitase. IscU2 also restored the aconitase activity in air-exposed M. acetivorans cell lysate. These biochemical results demonstrate that IscS2 is a cysteine desulfurase and that IscU2 is

Author: Kliefoth, Michael et al.; Genre: Journal Article; Published in Print: 2012-02; Keywords: Methanosarcina acetivoran; Carbon monoxide; Acclimation; Aldehyde dehydrogenase; Sensing; Regulation; Title: Genetic analysis of MA4079, an aldehyde dehydrogenase homolog, in Methanosarcina acetivorans

ID MEACE1_1_PE76 STANDARD; PRT; 348 AA. AC MEACE1_1_PE76; 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 (MEACE1_1.PE76). OS METHANOSARCINA ACETIVORANS C2A. OC Archaea; Euryarchaeota; Methanomicrobia; Methanosarcinales; OC Methanosarcinaceae; Methanosarcina. OX NCBI_TaxID=188937; RN [0] RP -.; RG -.; RL -.; CC -!- SEQ. DATA ORIGIN: Translated from the HOGENOM CDS MEACE1_1.PE76. CC Methanosarcina acetivorans C2A chromosome, complete genome. CC Genomes CC -!- GENE_FAMILY: HOG000224730 [ FAMILY / ALN / TREE ] DR HOGENOMDNA; MEACE1_1.PE76; -. SQ SEQUENCE 348 AA; UNKNOWN MW; UNKNOWN CRC64; MMFTSESDLM RTIDWNEESN SVVLVDQTLL PQEYKVIECK TLSSLCEAIK SLRIRGAPAL GAAGGFGIAL AASLSGAKDL ESMTRDLKVA AKALKSTRST AVNLGWGVDR VLNAISDAFD VQGARDIALQ EARDIAEEDI ATNKLIGKYG TKFLKDGDSV LTHCNAGRLA CVDWGTALGV VRSAIEEGKE IKVIACETRP LNQGSRITTW ELMQDKIPVT LIADSMAGWA MHQGLVDSVL VGADRITQDV VFNKIGTYTH SILAKEHEIP FYVAAPISTF DFKGWEGSVK IEMRDPDELR ...

General Information: Methanosarcina mazei Go1 (DSM 3647) was isolated from an anaerobic sewage digester in Germany. Anaerobic methane-producing archeon. This organism is a strictly anaerobic, non-motile, methane-producing archaeon. This organism can also aggregate forming large irregular shaped clumps of cells. Occasionally these aggregates can grow to 1000 microns or more in diameter. Growth occurs at pH 5.5-8.0, with optimum growth at pH 6.8-7.2. Growth occurs at pH 5.5-8.0, with optimum growth at pH 6.8-7.2. Can be found in decaying leaf litter, garden soil, sewage treatment sludge digestors, black mud, feces of herbivores and other urban waste and sewage products. ...

Authors: Gayen, Shovanlal; Vivekanandan, Subramanian; Biukovic, Goran; Gruber, Gerhard; Yoon, Ho Sup. Citation: Gayen, Shovanlal; Vivekanandan, Subramanian; Biukovic, Goran; Gruber, Gerhard; Yoon, Ho Sup. 1H, 13C, and 15N resonance assignments of subunit F of the A(1)A (O) ATP synthase from Methanosarcina mazei Go1 Biomol. NMR Assignments 1, 23-25 (2007).. Assembly members: ...

ID DNLI2_METMA Reviewed; 568 AA. AC Q8PTK1; DT 10-OCT-2002, integrated into UniProtKB/Swiss-Prot. DT 10-OCT-2002, sequence version 1. DT 25-OCT-2017, entry version 102. DE RecName: Full=DNA ligase 2 {ECO:0000255,HAMAP-Rule:MF_00407}; DE EC=6.5.1.1 {ECO:0000255,HAMAP-Rule:MF_00407}; DE AltName: Full=Polydeoxyribonucleotide synthase [ATP] 2 {ECO:0000255,HAMAP-Rule:MF_00407}; GN Name=lig2 {ECO:0000255,HAMAP-Rule:MF_00407}; GN OrderedLocusNames=MM_2714; OS Methanosarcina mazei (strain ATCC BAA-159 / DSM 3647 / Goe1 / Go1 / OS JCM 11833 / OCM 88) (Methanosarcina frisia). OC Archaea; Euryarchaeota; Methanomicrobia; Methanosarcinales; OC Methanosarcinaceae; Methanosarcina. OX NCBI_TaxID=192952; RN [1] RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=ATCC BAA-159 / DSM 3647 / Goe1 / Go1 / JCM 11833 / OCM 88; RX PubMed=12125824; RA Deppenmeier U., Johann A., Hartsch T., Merkl R., Schmitz R.A., RA Martinez-Arias R., Henne A., Wiezer A., Baeumer S., Jacobi C., RA Brueggemann H., Lienard T., ...

ID Q8PWC0_METMA Unreviewed; 498 AA. AC Q8PWC0; DT 01-OCT-2002, integrated into UniProtKB/TrEMBL. DT 01-OCT-2002, sequence version 1. DT 07-JUN-2017, entry version 83. DE SubName: Full=Type I restriction-modification system specificity subunit {ECO:0000313,EMBL:AAM31365.1}; DE EC=2.1.1.72 {ECO:0000313,EMBL:AAM31365.1}; GN OrderedLocusNames=MM_1669 {ECO:0000313,EMBL:AAM31365.1}; OS Methanosarcina mazei (strain ATCC BAA-159 / DSM 3647 / Goe1 / Go1 / OS JCM 11833 / OCM 88) (Methanosarcina frisia). OC Archaea; Euryarchaeota; Methanomicrobia; Methanosarcinales; OC Methanosarcinaceae; Methanosarcina. OX NCBI_TaxID=192952 {ECO:0000313,EMBL:AAM31365.1, ECO:0000313,Proteomes:UP000000595}; RN [1] {ECO:0000313,EMBL:AAM31365.1, ECO:0000313,Proteomes:UP000000595} RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=ATCC BAA-159 / DSM 3647 / Goe1 / Go1 / JCM 11833 / OCM 88 RC {ECO:0000313,Proteomes:UP000000595}; RX PubMed=12125824; RA Deppenmeier U., Johann A., Hartsch T., Merkl R., Schmitz R.A., RA ...

Methanogenesis, the biological production of methane, plays a pivotal role in the global carbon cycle and contributes significantly to global warming. The majority of methane in nature is derived from acetate. Here we report the complete genome sequence of an acetate-utilizing methanogen, Methanosarcina acetivorans C2A. Methanosarcineae are the most metabolically diverse methanogens, thrive in a broad range of environments, and are unique among the Archaea in forming complex multicellular structures. This diversity is reflected in the genome of M. acetivorans. At 5,751,492 base pairs it is by far the largest known archaeal genome. The 4524 open reading frames code for a strikingly wide and unanticipated variety of metabolic and cellular capabilities. The presence of novel methyltransferases indicates the likelihood of undiscovered natural energy sources for methanogenesis, whereas the presence of single-subunit carbon monoxide dehydrogenases raises the possibility of nonmethanogenic growth. ...

Abstract: The redox regulation of proteins via reversible dithiol/disulfide exchange reactions involves the thioredoxin system, which is composed of a reductant, a thioredoxin reductase (TR), and thioredoxin (Trx). In the pyridine nucleotide-dependent Trx reduction pathway, reducing equivalents, typically from reduced nicotinamide adenine dinucleotide phosphate (NADPH), are transferred from NADPH-TR (NTR) to Trx and, in turn, to target proteins, thus resulting in the reversible modification of the structural and functional properties of the targets. NTR enzymes contain three functional sites: an NADPH binding pocket, a non-covalently bound flavin cofactor, and a redox-active disulfide in the form of CxxC. With the aim of increasing our knowledge of the thioredoxin system in archaea, we here report the high-resolution crystal structure of NTR from the methane-generating organism Methanosarcina mazei strain Gö1 (MmNTR) at 2.6 Å resolution. Based on the crystals presently described, MmNTR assumes ...

All the other methanogens can utilize no more than two methanogenic substrates and possess a single pathway for methanogenesis. Methanosarcina, on the other hand, has all three known pathways for methanogenesis and can utilize no less than nine methanogenic substrates. M. barkeri and M. mazei are autotrophic, but M. acetivorans is not. It also has a number of distinct morphological forms including single cells with and without a cell envelope, as well as multicellular packets and lamina. The packets and lamina showed internal morphological diversity, indicating possible cell differentiation. The fact that cells in the lamina secrete different extracellular material gives light to possible cell specialization as well. They are coccoid and have cell walls of protein, often having an external wall of a heteropolysaccharide. Most Methanosarcina spp. are surrounded by a polymeric network of methanochondroitin that is external to an S-layer. The term matrix has been proposed to describe this ...

While ubiquitous among Archaea, and common in bacteria, the S-layers of diverse organisms have unique structural properties, including symmetry and unit cell dimensions, due to fundamental differences in their constituent building blocks.[14] Sequence analyses of S-layer proteins have predicted that S-layer proteins have sizes of 40-200 kDa and may be composed of multiple domains some of which may be structurally related. Since the first evidence of a macromolecular array on a bacterial cell wall fragment in the 1950s[15] S-layer structure has been investigated extensively by electron microscopy and medium resolution images of S-layers from these analyses has provided useful information on overall S-layer morphology. High-resolution structures of an archaeal S-layer protein (MA0829 from Methanosarcina acetivorans C2A) of the Methanosarcinales S-layer Tile Protein family and a bacterial S-layer protein (SbsB), from Geobacillus stearothermophilus PV72, have recently been determined by X-ray ...

A known 40 kbp bacteriophage insertion [2586000-2626000] is, surprisingly, not among the genomic fragments selected in the SWGB using this filter. Although the prophage is still perceptible on the Gene Map view (see a figure in the supplementary help web-pages), the OU parameters of the region do not differ markedly enough from the core sequence to be isolated automatically as a horizontally transferred region.. As the SWGB uses parameters that are based on comparison of local fragments to the global genomic average, strains with abundant insertions of homogenous DNA can confound this form of analysis. One example is the Methanosarcina acetivorans C2A genome which is composed of an estimated 25% of putatively horizontally acquired DNA, one of the highest amounts discovered to date [11]. As a result of these insertions, the genomic signature has been strongly influenced, resulting in a large amount of scatter and a poorly defined core genome on the plots. On the other hand, this type of ...

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Our team has designed and built the GeoBioCell -- a microfluidic gradient chamber created by etching a 1200-well array and boundary channels into a silicon wafer using photolithography methods. Archaea Methanosarcina acetivorans and Bacteria Escherichia coli were chosen as the model organisms to evaluate, and constraint conditions were created by establishing a concentration gradient within the GeoBioCell of an inhibitory antibiotic, i.e., puromycin for M. acetivorans and ciprofloxacin for E. coli. Cell growth was evaluated in batch systems for both M. acetivorans and E. coli in the presence of the antibiotic to determine the minimum inhibitory concentrations (MICs). The GeoBioCell design and results of E. coli growth in the presence of a ciprofloxacin are illustrated in Figure 1. Substrates (LB media, dissolved O2) were continuously delivered to the boundary channels of the GeoBioCell, while ciprofloxacin was continuously delivered to only the bottom boundary channel to establish a ...

The H+-translocating F420H2 Dehydrogenase (F420H2DH) Family (TC# 3.D.9) is a member of the Na+ transporting Mrp superfamily. A single F420H2 dehydrogenase (also referred to as F420H2:quinol oxidoreductase) from the methanogenic archaeon, Methanosarcina mazei Gö1, has been shown to be a redox driven proton pump. The F420H2DH of M. mazei has a molecular size of about 120 kDa and contains Fe-S clusters and FAD. A similar five-subunit enzyme has been isolated from Methanolobus tindarius. The sulfate-reducing Archaeoglobus fulgidus (and several other archaea) also have this enzyme. Reduction of 2-hydroxyphenazine by F420H2DH is accompanied by the translocation of 1 H+ per 2 electrons transferred. The overall vectorial reaction catalyzed by F420H2DH is Reduced donor (2e−) + H+ (in) ⇌ oxidized acceptor (2e−) + H+ (out) Methanomassiliicoccus luminyensis has been isolated from the human gut and requires H2 and methanol or methylamines to produce methane. The organism lacks cytochromes, indicating ...

Our approach is based on the assumption that proteins abundant in thermophilic and rare in mesophilic genomes are the most attractive targets for further biochemical investigations to understand the physiology specific for thermophiles in more detail. Consequently, our first goal was to identify such candidates via bioinformatic methods and as the most suitable protein THEP1 was selected for this study. The high ranking thermophile-specificity of THEP1 among procaryotes may be explained by an essential physiological role in thermophiles that is of no functional relevance for almost all mesophilic microorganisms. As an alternative explanation, a function also present in mesophilic organisms could be carried out by a protein that was not able to adapt to higher temperatures and in the course of convergent evolution, THEP1 could have taken over this particular function. Methanosarcina acetivorans str. C2A is the only mesophilic organism containing THEP1 (MA3402). Since the genome of M. acetivorans ...

Methanosarcina HMb protein: MW 11 kDa, 93 amino acid residues; acid-soluble component of Methananosarcina barkeri nucleoprotein complex; amino acid sequence given in first source

Authors: Barnwal, Ravi Pratap. Citation: Barnwal, Ravi; Agarwal, Geetika; Sharma, Yogendra; Chary, Kandala. Complete backbone assignment of a Ca2+-binding protein of the betagamma-crystallin superfamily from Methanosarcina acetivorans, at two denaturant concentrations Biomol. NMR Assignments 3, 107-110 (2009).. Assembly members: ...

Authors: Barnwal, Ravi Pratap; Agarwal, Geetika P; Chary, Kandala VR. Citation: Barnwal, Ravi; Agarwal, Geetika; Sharma, Yogendra; Chary, Kandala. Complete backbone assignment of a Ca2+-binding protein of the betagamma-crystallin superfamily from Methanosarcina acetivorans, at two denaturant concentrations Biomol. NMR Assignments 3, 107-110 (2009).. Assembly members: ...

A distinctive whiff of sulfides provided an unexpected dividend for then-graduate student James Moran while he was working with a carbon monoxide-metabolizing archeal methanogen Methanosarcina acetovorans. Now at McMaster University in Hamilton, Ontario, Canada, Moran was a graduate student with Christopher House of Pennsylvania State University at University Park.

This model describes a subfamily of the B12 binding domain (PF02607, PF02310) proteins. Members of the seed alignment include corrinoid proteins specific to four different, mutally non-homologous enzymes of the genus Methanosarcina. Three of the four cognate enzymes (trimethylamine, dimethylamine, and monomethylamine methyltransferases) all have the unusual, ribosomally incorporated amino acid pyrrolysine at the active site. All act in systems in which a methyl group is transferred to the corrinoid protein to create methylcobalamin, from which the methyl group is later transferred elsewhere ...

Branched five carbon (C 5) alcohols are attractive targets for microbial production due to their desirable fuel properties and importance as platform chemicals. In this study, we engineered a heterologous isoprenoid pathway in E. coli for the high-yield production of 3-methyl-3-buten-1-ol, 3-methyl-2-buten-1-ol, and 3-methyl-1-butanol, three C 5 alcohols that serve as potential biofuels. We first constructed a pathway for 3-methyl-3-buten-1-ol, where metabolite profiling identified NudB, a promiscuous phosphatase, as a likely pathway bottleneck. We achieved a 60% increase in the yield of 3-methyl-3-buten-1-ol by engineering the Shine-Dalgarno sequence of nudB, which increased protein levels by 9-fold and reduced isopentenyl diphosphate (IPP) accumulation by 4-fold. To further optimize the pathway, we adjusted mevalonate kinase (MK) expression and investigated MK enzymes from alternative microbes such as Methanosarcina mazei. Next, we expressed a fusion protein of IPP isomerase and the ...

A highly efficient method for chromosomal integration of cloned DNA into Methanosarcina spp. was developed utilizing the site-specific recombination system from the Streptomyces phage PhiC31. Host strains expressing the PhiC31 integrase gene and carrying an appropriate recombination site can be transformed with non-replicating plasmids carrying the complementary recombination site at efficiencies similar to those obtained with self-replicating vectors. We have also constructed a series of hybrid promoters that combine the highly expressed M. barkeri PmcrB promoter with binding sites for the tetracycline-responsive, bacterial TetR protein. These promoters are tightly regulated by the presence or absence of tetracycline in strains that express the tetRgene. The hybrid promoters can be used in genetic experiments to test gene essentiality by placing a gene of interest under their control. Thus, growth of strains with tetR-regulated essential genes becomes tetracycline-dependent. A series of plasmid ...

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Methanosarcina barkeri is the most fundamental species of the genus Methanosarcina, and their properties apply generally to the genus Methanosarcina. Methanosarcina barkeri can produce methane anaerobically through different metabolic pathways. M. barkeri can subsume a variety of molecules for ATP production, including methanol, acetate, methylamines, and different forms of hydrogen and carbon dioxide. Although it is a slow developer and is sensitive to change in environmental conditions, M. barkeri is able to grow in a variety of different substrates, adding to its appeal for genetic analysis. Additionally, M. barkeri is the first organism in which the amino acid pyrrolysine was found. Furthermore, two strains of M. barkeri, M. b. Fusaro and M. b. MS have been identified to possess an F-type ATPase (unusual for archaea, but common for bacteria, mitochondria and chloroplasts) along with an A-type ATPase. The fusaro strain of M. barkeri was found in mud samples taken from Lake Lago del Fusaro, a ...

Archaea is a peer-reviewed, Open Access journal that publishes original research articles as well as review articles dealing with all aspects of archaea, including environmental adaptation, enzymology, genetics and genomics, metabolism, molecular biology, molecular ecology, phylogeny, and ultrastructure. Published since 2002, Archaea provides a unique venue for exchanging information about these extraordinary prokaryotes.

The acetate kinase variants reported here all exhibited severalfold increases in the Km for acetate, supporting the hypothesized roles for the hydrophobic pocket residues Val93, Leu122, Phe179, and Pro232 as important contributors to acetate binding. The progressive increase in Km for acetate with a decrease in the side chain volume at position 93 supports the hypothesis that there is a decrease in hydrophobic interactions with acetate as the pocket size increases. Furthermore, the overall hydrophobicity of the pocket is decreased in the variants. Attempts to measure the Kd values of the variants for acetate and acetyl phosphate have been unsuccessful so far due to the upper solubility limit of acetate kinase (20 μM) relative to the Km values of the variants for acetate and acetyl phosphate, which were as high as 39.5 and 1.19 mM, respectively. The recently reported crystal structure of acetate kinase from M. thermophila containing acetate shows the methyl group located in the hydrophobic ...

The noncanonical amino acid S-allyl cysteine (Sac) is one of the major compounds of garlic extract and exhibits a range of biological activities. It is also a small bioorthogonal alkene tag capable of undergoing controlled chemical modifications, such as photoinduced thiol-ene coupling or Pd-mediated deprotection. Its small size guarantees minimal interference with protein structure and function. Here, we report a simple protocol efficiently to couple in-situ semisynthetic biosynthesis of Sac and its incorporation into proteins in response to amber (UAG) stop codons. We exploited the exceptional malleability of pyrrolysyl-tRNA synthetase (PylRS) and evolved an S-allylcysteinyl-tRNA synthetase (SacRS) capable of specifically accepting the small, polar amino acid instead of its long and bulky aliphatic natural substrate. We succeeded in generating a novel and inexpensive strategy for the incorporation of a functionally versatile amino acid. This will help in the conversion of orthogonal ...

CofE homologs are found in the genomes of all organisms currently known to produce coenzyme F420. These include not only the methanogenic archaea but also nonmethanogenic archaea such as Halobacteria and Archaeoglobus (see MF_01258), as well as some eubacteria such as Streptomyces, Mycobacterium, Nocardia, and Thermobifida (this rule, MF_01259). The length of the polyglutamate tail varies in different organisms. According to PubMed:12867481 (Graupner et al., 2003) and PubMed:11479701 (Bair et al., 2001): Analyses of the F420s present in Methanococcus jannaschii have shown that these cells contain a series of gamma-glutamyl-linked F420s capped with a single, terminal alpha-linked L-glutamate. The predominant form of F420 was designated as alpha-F420-3 and represented 86% of the F420s in these cells. Analyses of Methanosarcina thermophila, Methanosarcina barkeri, Methanobacterium thermoautotrophicum, Archaeoglobus fulgidus, and Mycobacterium smegmatis showed that they contained only ...

RN [1] RM 96134964 RT Characterization of the cdhD and cdhE genes encoding subunits of the corrinoid/iron-sulfur enzyme of the CO dehydrogenase complex from Methanosarcina thermophila. RA Maupin-Furlow J, Ferry JG RL J Bacteriol 1996 Jan;178(2):340-6 RN [2] RT Cloning and expression of the gene cluster encoding key proteins involved in acetyl-CoA synthesis in Clostridium thermoaceticum: CO dehydrogenase, the corrinoid/Fe-S protein, and methyltransferase. RA Roberts DL, James-Hagstrom JE, Garvin DK, Gorst CM, Runquist JA, Baur JR, Haase FC, Ragsdale SW RL Proc Natl Acad Sci U S A 1989 Jan;86(1):32-6 SE TIGR GA hmmls AL clustalw, belvu DR HAMAP; MF_01135; 9 of ...

The research objectives of the project are to develop genetic and molecular techniques that will permit the detection, isolation, and cloning of genes that are regulated during acetate catabolism. These studies should provide a firm basis for understanding the regulation of acetate utilization in the methanogen, Methanosarcina acitivorans. We have concentrated on three areas of study in the first year of the contract. They are; 1)development of cell plating methods for the methanosarcina, 2) screening and isolation of plasmids from the acetogenic methanogens, and 3) construction of gene libraries for M. acetivorans. It is anticipated that techniques developed in these studies will facilitate genetic study of other methanogenic organisms. Keywords: Archaebacteria, Methanogens, Acetate Utilization, Genetic Regulation, Plasmids, Nutrition.*ACETATES

Abstract : MC1 is the most abundant architectural protein present in Methanosarcina thermophila CHTI55 in laboratory growth conditions and is structurally unrelated to other DNA-binding proteins. MC1 functions are to shape and to protect DNA against thermal denaturation by binding to it. Therefore, MC1 has a strong affinity for any double-stranded DNA. However, it recognizes and preferentially binds to bent DNA, such as four-way junctions and negatively supercoiled DNA minicircles. Combining NMR data, electron microscopy data, biochemistry, molecular modelisation and docking approaches, we proposed recently a new type of DNA/protein complex, in which the monomeric protein MC1 binds on the concave side of a strongly bent 15 base pairs DNA. We present here the NMR chemical shifts assignments of each partner in the complex, 1H 15N MC1 protein and 1H 13C 15N bent duplex DNA, as first step towards the first experimental 3D structure of this new type of DNA/protein complex.. ...

2ZIN: Multistep Engineering of Pyrrolysyl-tRNA Synthetase to Genetically Encode N(varepsilon)-(o-Azidobenzyloxycarbonyl) lysine for Site-Specific Protein Modification

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 ...

Forever Active Pro-B blends six strains of friendly bacteria, selected and engineered for their ability to reach the intended destination of the large intestine. Forever Active Pro-B is the perfect high-quality friendly bacteria supplement to complement your gut flora and assist with your diet and lifestyle goals. Each strain goes through thorough testing and has been selected for its ability to bypass stomach acid for optimal delivery into the intestines. Forever Active Pro-B does not require refrigeration, but to ensure maximum benefits the capsules are stored in unique packaging that controls moisture and protects the goodness captured in each supplement. The formula is also free from allergens and suitable for vegetarians. - Promotes a healthy digestive system- Able to bypass stomach acid for optimal delivery within the intestines- Contains 6 synergistic strains including clinically studied Lactobacillus rhamnosus- Over 8 billion CFU- Strains cryo protected for potency and shelf stability- Soy and

Hypertension, additionally called hypertension, is a dangerous and potentially life-threatening medical situation. While the remedy might help the patients, this antithyroid medication for Graves Disease is only for the signs and never precisely for the basis reason for it. Sadly, for many patients the disease will still return they usually will have to go through the identical therapy over again.. Extracting salicin from herbs was considered to be expensive and time-consuming, so an artificial salicylic acid version was developed in Germany in 1852 and shortly became the therapy of alternative (salicin is transformed in the body to salicylic acid).. By blocking the calcium channels, these medicine cause the vessels to calm down, consequently blood strain goes down. Using the identical physician with experience with Autism and combining medications might make the search for a physician a little extra difficult.. Most of these over-the-counter heartburn medications have warnings on their labels ...

However, we are pros, and as such, we decided to experiment with taste, terpenes and cookie variety to see if we could make the perfect experience even more perfecter. Armed with a vaporizer in the basement of a private club, we filled bag after bag while chewing on four of the five most popular Girl Scout cookies, plus a gluten-free cookie no one should ever order, to determine which strain goes best with each cookie. We did it for you, dear reader. Only for you.. ...

As a member of the wwPDB, the RCSB PDB curates and annotates PDB data according to agreed upon standards. The RCSB PDB also provides a variety of tools and resources. Users can perform simple and advanced searches based on annotations relating to sequence, structure and function. These molecules are visualized, downloaded, and analyzed by users who range from students to specialized scientists.

If AOM Archaea oxidize methane by reverse methanogenesis, coenzyme F430 likely catalyzes the first step. Culture studies of methanogenic Archaea that can carry out trace oxidation of methane provide supporting evidence for a reversed methanogenesis biochemical pathway. The methanogen Methanosarcina acetovorines was shown to oxidize trace amounts of methane to CO2 (Moran et al., 2006) as documented by observations that 13C-labeled methane became incorporated into CO2. Studies of Methanothermobacter marburgensis in pure culture demonstrated the last step in methanogenesis is also the first step in methane oxidation (Scheller et al., 2010) by the incorporation of 13C-labeled methane into methyl-coenzyme M (2-mercaptoethanesulfonate) catalyzed by coenzyme F430. Genetic evidence from environmental samples provides additional support for reverse methanogenesis during AOM. Hallam et al., (2004) found genes that code for the enzymes used in methanogenesis, including for the last step, in ANME-1 and ...

Accepted name: cysteate synthase. Reaction: O-phospho-L-serine + sulfite = L-cysteate + phosphate. Other name(s): sulfite:O-phospho-L-serine sulfotransferase (phosphate-hydrolysing, L-cysteate-forming).. Systematic name: sulfite:O-phospho-L-serine sulfonotransferase (phosphate-hydrolysing, L-cysteate-forming).. Comments: A pyridoxal-phosphate protein. It is highly specific for O-phospho-L-serine and sulfite. The reaction proceeds through a dehydroalanine (2-aminoacrylic acid) intermediate. The enzyme from Methanosarcina acetivorans is evolutionarily related to threonine synthase (EC 4.2.3.1), but the reaction is more similar to that of O-phosphoserine sulfhydrylase (EC 2.5.1.65).. Links to other databases: BRENDA, EXPASY, KEGG, Metacyc, CAS registry number: References:. 1. Graham, D.E., Taylor, S.M., Wolf, R.Z. and Namboori, S.C. Convergent evolution of coenzyme M biosynthesis in the Methanosarcinales: cysteate synthase evolved from an ancestral threonine synthase. Biochem. J. 424 (2009) ...

Ultrastructural examinations were performed on biofilms from eight anaerobic fixed-bed reactors filled with various packing materials and operated on fresh swine waste. By using light, UV, scanning, and transmission electron microscopy, the distribution of a diverse microbial population composed of bacteria and a few yeasts was determined. This is the first time that the ultrastructure of in situ anaerobic digestor biofilms has been reported. A large number of methanogenic bacteria were identified by their fluorescence under 420 nm of radiation. Of these, two morphologically distinct types were most prevalent in the films. Methanothrix spp. was present in high numbers at the film surface, whereas Methanosarcina spp. were commonly embedded in the lower regions of the of the film. Inhabitants of the film were surrounded by an exopolysaccharide matrix that was very dense toward the base. An extensive network of channels was observed throughout the matrix that may facilitate gas and nutrient ...

Pyrrolysine, the 22nd cotranslationally inserted amino acid, was found in the Methanosarcina barkeri monomethylamine methyltransferase protein in a position that is encoded by an in-frame UAG stop codon in the mRNA. M. barkeri encodes a special amber suppressor tRNA (tRNA(Pyl)) that presumably recog …

The binding of MC1 protein, the major chromosomal protein of the archaebacterium Methanosarcina sp. CHTI 55, to the region preceding the strongly expressed genes encoding methyl coenzyme reductase in a closely related micro-organism has been investigated. By gel retardation and DNAase I footprinting assays, we identified a preferential binding sequence in an open reading frame of unknown function. The large area of DNA protected against DNAase I is interrupted by a strong cleavage enhancement site on each strand. By circular permutation assays, we showed that the DNA bends upon MC1 binding. Furthermore we observed that the presence of a sequence outside the binding site can induce an unusual electrophoretic behaviour in some complexes. ...

The SCOP classification for the Monomethylamine methyltransferase MtmB superfamily including the families contained in it. Additional information provided includes InterPro annotation (if available), Functional annotation, and SUPERFAMILY links to genome assignments, alignments, domain combinations, taxonomic visualisation and hidden Markov model information.

All M. acetivorans strains (Additional file 1: Table S3) were routinely grown anaerobically as pre-cultures at 37 °C in an 80 % N2/19 % CO2/1 % H2 atmosphere with mild shaking in 10 mL HS medium [28] with 150 mM methanol as the carbon source. Cell growth was measured spectrophotometrically and direct cell counts were confirmed by staining cell cultures (often containing precipitates) with SYTO9 dye (Life Technologies, Carlsbad, CA, USA) and viewed microscopically using a bright-line hemocytometer (Hausser Scientific, Horsham, PA, USA) under phase-contrast and epifluorescence settings (Zeiss Axio Scope.A1, Germany). All 28-mL culture tubes (18 × 150 mm, Bellco Glass, Vinelanad, NJ, USA) were sealed by aluminum crimp seals. Plasmids were maintained with 2 µg/mL puromycin.. For long-term (ca., 40 days) growth on methane with low-density inocula (1 %), the M. acetivorans strains were grown in 5 mL HS medium in 28-mL culture tubes with additional electron acceptors at 37 °C with mild shaking; the ...

SWISS-MODEL Repository entry for P80521 (PORA_METBF), Pyruvate synthase subunit PorA. Methanosarcina barkeri (strain Fusaro / DSM 804)

1. Cup shaped; cuplike. 2. Relating to the c. cavity or acetabulum. [G. kotyle, a small cup, + eidos, appearance] * * * cot·y·loid kät əl .ȯid adj of or relating to an acetabulum * * * cot·y·loid (kotґə loid) [Gr. kotyloeides cup shaped] 1