Corrinoid-dependent ethyl-transfer catalyzed by the methanol:coenzyme M methyl transferase from Methanosarcina acetivorans DOI Creative Commons
Tejas Somvanshi, Jichen Bao, Silvan Scheller

et al.

Research Square (Research Square), Journal Year: 2024, Volume and Issue: unknown

Published: July 30, 2024

Abstract Corrinoid-dependent methyltransferases catalyze methyl-group transfer reactions in all domains of life. These enzymes are generally considered exclusive for C1-substrates (methyl-groups). However, Methanosarcina trace ethane production from ethanol has been demonstrated in vivo, which led to the hypothesis that corrinoid-dependent methanol specific promiscuous towards also accepting ethyl-groups. Here we show conversion amounts Methanosarcina acetivorans proceeds via known methanol-to-methane metabolism, involving methanol:5-hydroxybenzimidazolylcobamide methyltransferase (MtaB) and a corrinoid-containing methyl-accepting protein (MtaC), but ethyl groups instead methyl groups. We demonstrate three isozymes MtaB subunit corrinoid MtaC M. ethanol, granting microbe capacity promiscuity downstream Co-methyl-5-hydroxybenzimidazolylcobamide:2-mercaptoethanesulfonate (MtaA) methyl-coenzyme M reductase (Mcr). assessed ethyl-group efficiency each engineered chimeras combine 2 different MtaA subunits with 3 isoforms MtaCB together increase capability acetivorans. Demonstrating coenzyme can higher alkyl extends pool be metabolic networks.

Language: Английский

Physiological barriers for glucose utilization inMethanosarcina acetivorans DOI Creative Commons
Christian Sattler, Marcus Richter, Michael Rother

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Aug. 22, 2024

Methanogenesis is a key aspect of anaerobic biomass degradation, and thus global importance. While methanogenic archaea (methanogens) are ubiquitous in habitats, the range substrates they utilize very limited. Most methanogens able to grow chemolithotrophically with H 2 +CO , some clades can methylated compounds and/or acetate as well. Organotrophic like amino acids, lipids, nucleotides, or carbohydrates, not known support growth methanogens. This inability “upstream” intermediates degradation remarkable considering presence cellular metabolism that such could feed into. Here, we addressed question why model methanogen Methanosarcina acetivorans despite its gluconeogenic glycolytic capacity, unable glucose for methanogenesis growth. Complementing heterologously uptake facilitator allowed recombinant M. strain convert methane at low rate. However, was observed, neither energy source nor carbon source. Instead, methylotrophic transgenic impaired glucose-dependent fashion, which aggravated when also glucokinase heterologuously produced. Glucose-dependent inhibition coincided significant – microscopically visible accumulation intracellular carbohydrate. Since glucose-utilizing trait rapidly lost during growth, accumulating growth-inhibiting metabolites probably makes catabolism incompatible. Thus, extensive efforts development would be required enable direct utilization methanogenesis.

Language: Английский

Citations

1
Corrinoid-dependent ethyl-transfer catalyzed by the methanol:coenzyme M methyl transferase from Methanosarcina acetivorans DOI Creative Commons
Tejas Somvanshi, Jichen Bao, Silvan Scheller

et al.

Research Square (Research Square), Journal Year: 2024, Volume and Issue: unknown

Published: July 30, 2024

Abstract Corrinoid-dependent methyltransferases catalyze methyl-group transfer reactions in all domains of life. These enzymes are generally considered exclusive for C1-substrates (methyl-groups). However, Methanosarcina trace ethane production from ethanol has been demonstrated in vivo, which led to the hypothesis that corrinoid-dependent methanol specific promiscuous towards also accepting ethyl-groups. Here we show conversion amounts Methanosarcina acetivorans proceeds via known methanol-to-methane metabolism, involving methanol:5-hydroxybenzimidazolylcobamide methyltransferase (MtaB) and a corrinoid-containing methyl-accepting protein (MtaC), but ethyl groups instead methyl groups. We demonstrate three isozymes MtaB subunit corrinoid MtaC M. ethanol, granting microbe capacity promiscuity downstream Co-methyl-5-hydroxybenzimidazolylcobamide:2-mercaptoethanesulfonate (MtaA) methyl-coenzyme M reductase (Mcr). assessed ethyl-group efficiency each engineered chimeras combine 2 different MtaA subunits with 3 isoforms MtaCB together increase capability acetivorans. Demonstrating coenzyme can higher alkyl extends pool be metabolic networks.

Language: Английский

Citations

0