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Examination and reconstitution of the glycine betaine-dependent methanogenesis pathway from the obligate methylotrophic methanogen Methanolobus vulcani B1d

Creighbaum, Adam J.
http://orcid.org/0000-0002-2852-8400
http://rave.ohiolink.edu/etdc/view?acc_num=miami158754951585964

Abstract Details

2020, Doctor of Philosophy, Miami University, Microbiology.
Recent studies indicate that environmentally abundant quaternary amines (QAs) are a primary source for methanogenesis, yet the catabolic enzymes are unknown. We hypothesized that the methanogenic archaeon Methanolobus vulcani B1d metabolizes glycine betaine through a corrinoid-dependent glycine betaine:coenzyme M (CoM) methyl transfer pathway. The draft genome sequence of M. vulcani B1d revealed a gene encoding a predicted non-pyrrolysine MttB homolog (MV8460) with high sequence similarity to the glycine betaine methyltransferase encoded by Desulfitobacterium hafniense Y51. MV8460 catalyzes glycine betaine-dependent methylation of free cob(I)alamin indicating it is an authentic MtgB enzyme. Proteomic analysis revealed that MV8460 and a corrinoid binding protein (MV8465) were highly abundant when M. vulcani B1d was grown on glycine betaine relative to growth on trimethylamine. The abundance of a corrinoid reductive activation enzyme (MV10335) and a methylcorrinoid:CoM methyltransferase (MV10360) were significantly higher in GB-grown B1d lysates compared to other homologs. The glycine betaine:CoM pathway was fully reconstituted in vitro using recombinant MV8460, MV8465, MV10335, and MV10360. Demonstration of the complete glycine betaine:CoM pathway expands the knowledge of direct QA-dependent methylotrophy and establishes a model to identify additional ecologically relevant anaerobic quaternary amine pathways.
Donald Ferguson, Jr. (Advisor)
Annette Bollmann (Committee Member)
Xin Wang (Committee Member)
Rachael Morgan-Kiss (Committee Member)
Richard Page (Committee Member)
170 p.
Microbiology
Methanogenesis; methane; C1; glycine betaine; quaternary amine; methanolobus; one-carbon; metabolism

Recommended Citations

Citations

  • Creighbaum, A. J. (2020). Examination and reconstitution of the glycine betaine-dependent methanogenesis pathway from the obligate methylotrophic methanogen Methanolobus vulcani B1d [Doctoral dissertation, Miami University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=miami158754951585964

    APA Style (7th edition)

  • Creighbaum, Adam. Examination and reconstitution of the glycine betaine-dependent methanogenesis pathway from the obligate methylotrophic methanogen Methanolobus vulcani B1d. 2020. Miami University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=miami158754951585964.

    MLA Style (8th edition)

  • Creighbaum, Adam. "Examination and reconstitution of the glycine betaine-dependent methanogenesis pathway from the obligate methylotrophic methanogen Methanolobus vulcani B1d." Doctoral dissertation, Miami University, 2020. http://rave.ohiolink.edu/etdc/view?acc_num=miami158754951585964

    Chicago Manual of Style (17th edition)

miami158754951585964
248
© 2020, some rights reserved.Creative Commons License Examination and reconstitution of the glycine betaine-dependent methanogenesis pathway from the obligate methylotrophic methanogen Methanolobus vulcani B1d by Adam J. Creighbaum is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. Based on a work at etd.ohiolink.edu.
This open access ETD is published by Miami University and OhioLINK.