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Demethylation and degradation of phenylmethylethers by the sulfide-methylating homoacetogenic bacterium strain TMBS 4

Abstract

Biochemical studies on anaerobic phenylme-thylether cleavage by homoacetogenic bacteria have been hampered so far by the complexity of the reaction chain involving methyl transfer to acetyl-CoA synthase and subsequent methyl group carbonylation to acetyl-CoA. Strain TMBS 4 differs from other demethylating homoacetogenic bacteria in using sulfide as a methyl acceptor, thereby forming methanethiol and dimethylsulfide. Growing and resting cells of strain TMBS 4 used alternatitively CO2 as a precursor of the methyl acceptor CO for homoacetogenic acetate formation. Demethylation was inhibited by propyl iodide and reactivated by light, indicating involvement of a corrinoid-dependent methyltransferase. Strain TMBS 4 contained ca. 750 nmol g dry mass-1 of a corrinoid tentatively identified as 5-hydroxybenzimidazolyl cobamide. A photometric assay for measuring the demethylation activity in cell extracts was developed based on the formation of a yellow complex of Ti3+ with 5-hydroxyvanillate produced from syringate by demethylation. In cell extracts, the methyltransfer reaction from methoxylated aromatic compounds to sulfide or methanethiol depended on reductive activation by Ti3+. ATP and Mg2+ together greatly stimulated this reductive activation without being necessary for the demethylation reaction itself. The specific activity of the transmethylating enzyme system increased proportionally with protein concentration up to 3 mg ml-1 reaching a constant level of 20 nmol min-1 mg-1 at protein concentrations ≥ 10 mg ml-1. The specific rate of activation increased in a non-linear manner with protein concentration. Strain TMBS 4 degraded gallate, the product of sequential demethylations, to 3 acetate through the phloroglucinol pathway as found earlier with Pelobacter acidigallici.

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Abbreviations

BV:

benzyl viologen

CTAB:

cetyltrimethylammonium bromide

H4folate:

tetrahydrofolate

MOPS:

3-[N-morpholino]propanesulfonic acid

MV:

methyl viologen

NTA:

nitrilotriacetate

td :

doubling time

TMB:

3,4,5-trimethoxybenzoate

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Kreft, JU., Schink, B. Demethylation and degradation of phenylmethylethers by the sulfide-methylating homoacetogenic bacterium strain TMBS 4. Arch. Microbiol. 159, 308–315 (1993). https://doi.org/10.1007/BF00290912

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  • DOI: https://doi.org/10.1007/BF00290912

Key words

  • Methyl transfer
  • Demethylation
  • Methoxylated aromatic compounds
  • Ether cleavage
  • Corrinoids
  • Homoacetogenic bacteria
  • Phloroglucinol pathway
  • Dimethylsulfide