Cloning and expression of the genes encoding the propene monooxygenase from Xanthobacter, Py2

N. Y. Zhou, C. K. Chan Kwo Chion, D. J. Leak

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Xanthobacter Py2 grows on propene as sole carbon source, converting propene to propene oxide (epoxypropane) using an alkene-specific monooxygenase, as the first step in catabolism. Four mutants, NZ1-4, with a propene- propene oxide+ phenotype were isolated by 1-methyl-3-nitro-1-nitrosoguanidine mutagenesis or by enrichment with the suicide substrate vinylidene chloride, and were shown to have lost the ability to convert alkenes to epoxides. All four mutants were complemented by a number of clones of Xanthobacter Py2 chromosomal DNA in the broad-host-range cosmid pLAFR5, some of which appeared to be non-overlapping. Representatives of the different clones obtained were transferred into Xanthobacter autotrophicus JW33 and one, pNY2, the most frequently isolated clone, was shown to express an inducible, fully functional propene monooxygenase. Subcloning revealed that all four mutants were complemented by a 2.4-kb EcoRI-PstI fragment situated at one end of the cosmid insert. However, activity in X. autotrophicus JW33 could only be expressed from pNY2, containing the complete insert (25 kb), suggesting a large operon or some form of long-range control. pNY2 failed to express in E. coli. In X. autotrophicus JW33 [pNY2] at least three new polypeptides were evident after induction with propene compared with a control carrying only the cosmid pLAFR5.

Original languageEnglish
Pages (from-to)582-588
Number of pages7
JournalApplied Microbiology and Biotechnology
Volume44
Issue number5
DOIs
Publication statusPublished - 1 Jan 1996

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

Cloning and expression of the genes encoding the propene monooxygenase from Xanthobacter, Py2. / Zhou, N. Y.; Chan Kwo Chion, C. K.; Leak, D. J.

In: Applied Microbiology and Biotechnology, Vol. 44, No. 5, 01.01.1996, p. 582-588.

Research output: Contribution to journalArticle

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