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Archives of Microbiology

, Volume 159, Issue 4, pp 308–315 | Cite as

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

  • Jan-Ulrich Kreft
  • Bernhard Schink
Original Papers

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.

Key words

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

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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Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • Jan-Ulrich Kreft
    • 1
  • Bernhard Schink
    • 1
  1. 1.Fakultät für BiologieUniversität KonstanzKonstanzGermany

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