Skip to main content
Log in

Dithiol- and NAD-dependent degradation of epoxyalkanes by Xanthobacter Py2

  • Environmental Biotechnology
  • Original Paper
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

A broad range of epoxyalkanes was converted into the corresponding ketones by cell extracts of Xanthobacter Py2. Both 1,2- and 2,3-epoxyalkanes were degraded and in addition, the degradation of 2,3-epoxyalkanes in all cases was highly enantioselective. Conversion of a deuterium-labelled substrate indicated that the ketone product was probably formed indirectly via an hydroxy intermediate. Degradation of epoxyalkanes by Xanthobacter Py2 was dependent on both NAD and another, not yet identified, cofactor that was present in the low-molecular-mass fraction (LMF) of propene-grown cells. It is proposed that the LMF was involved in a reductive reaction step since it could be replaced by dithiothreitol (DTT) and various other dithiol compounds. Epoxyalkane-degrading activity was inhibited by the sulphhydryl blocking reagent N-ethylmaleimide (NEM). Inhibition by NEM and stimulation by LMF, DTT and other dithiols was effective only in the simultaneous presence of NAD.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Bont JAM de (1993) Bioformation of optically pure epoxides. Tetrahedron: Asymmetry 4:1331–1340

    Google Scholar 

  • Bont JAM de, Harder W (1978) Metabolism of ethylene by Mycobacterium E20. FEMS Microbiol Lett 3:89–93

    Google Scholar 

  • Bont JAM de, Dijken JP van, Ginkel CG van (1982) The metabolism of 1,2-propanediol by the propylene oxide utilizing bacterium Nocardia A60. Biochim Biophys Acta 714:465–470

    Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Carter SF, Leak DJ (1993) The isolation and genetic construction of cyclic epoxide degrading bacteria. In: Griengl H, Steiner W, Faber K, Preisz A (eds) Abstracts of the European Symposium on Biocatalysis, Graz, Austria, 12–17 Sept 1993, Federal Ministry for Science and Research, Vienna, p P-50

    Google Scholar 

  • Cleland WW (1964) Dithiothreitol, a new protective reagent for SH groups. Biochemistry 3:480–482

    CAS  PubMed  Google Scholar 

  • Dobson NA, Eglinton G, Krishnamurti M, Raphael RA, Willis RG (1961) Selective catalytic hydrogenation of acetylenes. Tetrahedron 16:16–24

    Google Scholar 

  • Fasco MJ, Preusch PC, Hildebrandt E, Suttie JW (1983) Formation of hydroxyvitamin K by vitamin K epoxide reductase of warfarin-resistant rats. J Biol Chem 258:4372–4380

    Google Scholar 

  • Ginkel CG van, Bont JAM de (1986) Isolation and characterization of alkene-utilizing Xanthobacter spp. Arch Microbiol 145:403–407

    Google Scholar 

  • Ginkel CG van, Welten HGJ, Bont JAM de (1986) Epoxidation of alkenes by alkene-grown Xanthobacter spp. Appl Microbiol Biotechnol 24:334–337

    Google Scholar 

  • Hartmans S, Smits JP, Werf MJ van der, Volkering F, Bont JAM de (1989) Metabolism of styrene oxide and 2-phenylethanol in the styrene-degrading Xanthobacter strain 124X. Appl Environ Microbiol 55:2850–2855

    Google Scholar 

  • Hechtberger P, Wirnsberger G, Mischitz M, Klempier N, Faber K (1993) Asymmetric hydrolysis of epoxides using an immobilized enzyme preparation from Rhodococcus sp. Tetrahedron: Asymmetry 4:1161–1164

    Google Scholar 

  • Jacobs MHJ, Wijngaard AJ van den, Pentenga M, Janssen DB (1991) Characterization of the epoxide hydrolase from an epichlorohydrin-degrading Pseudomonas sp. Eur J Biochem 202:1217–1222

    Google Scholar 

  • Kasai N, Tsujimura K, Unoura K, Suzuki T (1990) Degradation of 2,3-dichloro-1-propanol by a Pseudomonas sp. Agric Biol Chem 54:3185–3190

    Google Scholar 

  • Leak DJ, Aikens PJ, Seyed-Mahmoudian M (1992) The microbial production of epoxides. TIBTECH 10:256–261

    Google Scholar 

  • Lee JJ, Fasco MJ (1984) Metabolism of vitamin K and vitamin K 2,3-epoxidevia interaction with a common disulfide. Biochemistry 23:2246–2252

    Google Scholar 

  • Liptay-Reuter I, Dose K, Guenthner T, Wörner W, Oesch F (1985) Vitamin K epoxide reductase activity in the metabolism of epoxides. Biochem Pharmacol 34:2617–2620

    Google Scholar 

  • Ophem PW van, Beeumen J van, Duine JA (1992) NAD-linked, factor-dependent formaldehyde dehydrogenase or trimeric, zinc-containing, long-chain alcohol dehydrogenase from Amycolatopsis methanolica. Eur J Biochem 206:511–518

    Google Scholar 

  • Pasto DJ, Cumbo CC (1965) An improved methodfor the preparation of volatile epoxides. J Org Chem 30:1271–1272

    Google Scholar 

  • Pedragosa-Moreau S, Archelas A, Furstoss R (1993) Enantiocomplementary epoxide hydrolyses as a preparative access to both enantiomers of styrene oxide. J Org Chem 58:5533–5536

    Google Scholar 

  • Schurig V, Bürkle W (1982) Extending the scope of enantiomer resolution by complexation gas chromatography. J Am Chem Soc 104:7573–7580

    Google Scholar 

  • Silverman RB (1981) Chemical model studies for the mechanism of vitamin K epoxide reductase. J Am Chem Soc 103:5939–5941

    Google Scholar 

  • Steckbeck SR, Nelson JA, Spencer TA (1982) Enzymic reduction of an epoxide to an alcohol. J Am Chem Soc 104:893–895

    Google Scholar 

  • Torchinskii YuM (1974) The role of SH groups in enzymes. In: Torchinskii YuM (ed) Sulfhydryl and disulfide groups of proteins. Consultants Bureau, London, pp 185–198

    Google Scholar 

  • Weijers CAGM, Haan A de, Bont JAM de (1988a) Microbial production and metabolism of epoxides. Microbiol Sci 5:156–159

    Google Scholar 

  • Weijers CAGM, Haan A de, Bont JAM de (1988b) Chiral resolution of 2,3-epoxyalkanes by Xanthobacter Py2. Appl Microbiol Biotechnol 27:337–340

    Google Scholar 

  • Whitlon DS, Sadowski JA, Suttie JW (1978) Mechanism of coumarin action: significance of vitamin K epoxide reductase inhibition. Biochemistry 17:1371–1377

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Weijers, C.A.G.M., Jongejan, H., Franssen, M.C.R. et al. Dithiol- and NAD-dependent degradation of epoxyalkanes by Xanthobacter Py2. Appl Microbiol Biotechnol 42, 775–781 (1995). https://doi.org/10.1007/BF00171961

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00171961

Keywords

Navigation