Archives of Microbiology

, Volume 159, Issue 1, pp 16–20 | Cite as

Formation of d-3-hydroxybutyryl-coenzyme A by an acetoacetyl-coenzyme A reductase in Syntrophomonas wolfei subsp. wolfei

  • Dale A. Amos
  • Michael J. McInerney
Original Papers

Abstract

Cell-free extracts of Syntrophomonas wolfei subsp. wolfei synthesized d-(-)-3-hydroxybutyryl-coenzyme A (CoA) (the stereoisomer required for the synthesis of poly-β-hydroxyalkanoate) from acetoacetyl-CoA, but not crotonyl-CoA, and NAD(P)H. Ammonium sulfate fractionation and ion exchange chromatography separated an acetoacetyl-CoA reductase activity that formed d-(-)-3-hydroxybutyryl-CoA from the β-oxidation enzyme activity, l-(+)-3-hydroxyacyl-CoA dehydrogenase. The former activity was further purified by hydroxylapatite and affinity chromatography. The most pure acetoacetyl-CoA reductase preparations formed d-(-)-3-hydroxybutyryl-CoA from acetoacetyl-CoA and had high specific activities using either NADH or NADPH as the electron donor. Thus, S. wolfei makes d-(-)-3-hydroxybutyryl-CoA by an acetoacetyl-CoA reductase rather than by a d-isomer specific enoyl-CoA hydratase and the reducing equivalents required for PHA synthesis from acetoacetyl-CoA can be supplied from the NADH made during β-oxidation.

Key words

Poly-β-hydroxylalkanoate Methanogenesis Syntrophic Fatty acid metabolism Syntrophomonas wolfei 

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Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • Dale A. Amos
    • 1
  • Michael J. McInerney
    • 1
  1. 1.Department of Botany and MicrobiologyUniversity of OklahomaNormanUSA

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