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Title: In vitro methanol production from methyl coenzyme M using the Methanosarcina barkeri MtaABC protein complex

Authors:
 [1];  [1];  [1];  [2]; ORCiD logo [3];  [2];  [1]
  1. Dept. of Chemistry & Biochemistry, University of Delaware, Newark DE 19716
  2. Dept. of Chemical & Biomolecular Engineering, Delaware Biotechnology Inst., University of Delaware, Newark DE 19711
  3. Dept. of Chemical & Biomolecular Engineering, University of Delaware, Newark DE 19716
Publication Date:
Sponsoring Org.:
USDOE Advanced Research Projects Agency - Energy (ARPA-E)
OSTI Identifier:
1399795
Grant/Contract Number:
AR0000432
Resource Type:
Journal Article: Publisher's Accepted Manuscript
Journal Name:
Biotechnology Progress
Additional Journal Information:
Journal Volume: 33; Journal Issue: 5; Related Information: CHORUS Timestamp: 2017-10-17 01:31:03; Journal ID: ISSN 8756-7938
Publisher:
Wiley Blackwell (John Wiley & Sons)
Country of Publication:
United States
Language:
English

Citation Formats

Dong, Ming, Gonzalez, Tara D., Klems, Meghan M., Steinberg, Lisa M., Chen, Wilfred, Papoutsakis, Eleftherios T., and Bahnson, Brian J.. In vitro methanol production from methyl coenzyme M using the Methanosarcina barkeri MtaABC protein complex. United States: N. p., 2017. Web. doi:10.1002/btpr.2503.
Dong, Ming, Gonzalez, Tara D., Klems, Meghan M., Steinberg, Lisa M., Chen, Wilfred, Papoutsakis, Eleftherios T., & Bahnson, Brian J.. In vitro methanol production from methyl coenzyme M using the Methanosarcina barkeri MtaABC protein complex. United States. doi:10.1002/btpr.2503.
Dong, Ming, Gonzalez, Tara D., Klems, Meghan M., Steinberg, Lisa M., Chen, Wilfred, Papoutsakis, Eleftherios T., and Bahnson, Brian J.. Mon . "In vitro methanol production from methyl coenzyme M using the Methanosarcina barkeri MtaABC protein complex". United States. doi:10.1002/btpr.2503.
@article{osti_1399795,
title = {In vitro methanol production from methyl coenzyme M using the Methanosarcina barkeri MtaABC protein complex},
author = {Dong, Ming and Gonzalez, Tara D. and Klems, Meghan M. and Steinberg, Lisa M. and Chen, Wilfred and Papoutsakis, Eleftherios T. and Bahnson, Brian J.},
abstractNote = {},
doi = {10.1002/btpr.2503},
journal = {Biotechnology Progress},
number = 5,
volume = 33,
place = {United States},
year = {Mon Jun 12 00:00:00 EDT 2017},
month = {Mon Jun 12 00:00:00 EDT 2017}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record at 10.1002/btpr.2503

Citation Metrics:
Cited by: 1work
Citation information provided by
Web of Science

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  • The transformation of the methyl group of methanol into methyl coenzyme M proceeds with net retention of methyl group configuration and without significant racemization. This is consistent with a proposed mechanism in which the methyl group is transferred from methanol first to the cobalt of the corrinoid enzyme MT/sub 1/ and then to the sulfur of coenzyme M. This resembles the transfer of the methyl group of methyltetrahydrofolate to homocysteine, catalyzed by the B/sub 12/-dependent methionine synthase from E. coli, which the authors have demonstrated also occurs with net retention of methyl group configuration. Both reactions pose the same questionmore » of how a relatively inert bond, the C-O bond of methyltetrahydrofolate in the case of methionine synthase, is cleaved in the transfer of a methyl group.« less
  • Extracts of Methanosarcina barkeri possess a specific methyltransferase that catalyzes the transfer of the methyl group of methanol to 2-mercaptoethanesulfonic acid. Over a fourfold range in added 2-mercaptoethanesulfonic acid, the formation of 2-(methylthio)ethanesulfonic acid exhibited a 1:1 ratio to 2-mercaptoethanesulfonic acid added. This reaction required adenosine 5'-triphosphate; a maximal ratio (mole/mol) of 85 methyl groups was transferred per adenosine 5'-triphosphate added. The methyltransferase was found in extracts of methanol-grown cells as well as in extracts of hydrogen-grown cells. In extracts of cells grown on either substrate, 2-(methylthio)ethanesulfonic acid was formed from added methanol or methylamine but not from acetate.
  • The transfer of the methyl group of acetate to coenzyme M (2-mercaptoethanesulfonic acid; HS-CoM) during the metabolism of acetate to methane was investigated in cultures of Methanosarcina strain TM-1. The organism metabolized CD/sub 3/COO/sup -/ to 83% CD/sub 3/H and 17% CD/sub 2/H/sub 2/ and produced no CDH/sub 3/ or CH/sub 4/. The isotopic composition of coenzyme M in cells grown on CD/sub 3/COO/sup -/ was analyzed with a novel gas chromatography-mass spectrometry technique. The cells contained CD/sub 3/-S-CoM and CD/sub 2/H-S-CoM) in a proportion similar to that of CD/sub 3/H to CD/sub 2/H/sub 2/. These results, in conjunction withmore » a report that extracts of acetate-grown strain TM-1 contain high levels of CH/sub 3/-S-CoM methylreductase, indicate that CH/sub 3/-S-CoM is an intermediate in the metabolism of acetate to methane in this organism.« less
  • The purified nickel-containing CO dehydrogenase complex isolated from methanogenic Methanosarcina thermophila grown on acetate is able to catalyze the exchange of (1-{sup 14}C) acetyl-coenzyme A (CoA) (carbonyl group) with {sup 12}CO as well as the exchange of (3'-{sup 32}P)CoA with acetyl-CoA. Kinetic parameters for the carbonyl exchange have been determined: Km (acetyl-CoA) = 200 microM, Vmax = 15 min-1. CoA is a potent inhibitor of this exchange (Ki = 25 microM) and is formed under the assay conditions because of a slow but detectable acetyl-CoA hydrolase activity of the enzyme. Kinetic parameters for both exchanges are compared with those previouslymore » determined for the acetyl-CoA synthase/CO dehydrogenase from the acetogenic Clostridium thermoaceticum. Collectively, these results provide evidence for the postulated role of CO dehydrogenase as the key enzyme for acetyl-CoA degradation in acetotrophic bacteria.« less
  • Methanol:5-hydroxybenzimidazolylcobamide methyltransferase from Methanosarcina barkeri has been purified to approximately 90% homogeneity by ion-exchange chromatography on DEAE-cellulose and QAE-A50 Sephadex columns. The molecular weight, estimated by gel electrophoresis, was found to be 122,000, and the enzyme contained two different subunits with molecular weights of 34,000 and 53,000, which indicates an ..cap alpha../sub 2/..beta.. structure. The enzyme contains three or four molecules of 5-hydroxybenzimidazolylcobamide, which could be removed by treatment of the enzyme with 2-mercaptoethanol or sodium dodecyl sulfate. In both cases the enzyme dissociated into its subunits. For stability, the enzyme required the presence of divalent cations such as Mg/supmore » 2 +/, Mn/sup 2 +/, Sr/sup 2 +/, Ca/sup 2 +/, or Ba/sup 2 +/. ATP, GTP, or CTP was needed in a reductive activation process of the enzyme. This activations was brought about by a mixture of H/sub 2/, ferredoxin, and hydrogenase, but also by CO, which is thought to reduce the corrinoid chemically. The CO dehydrogenase-like activity of the methyltransferase is discussed.« less