Evidence for involvement of copper ions and redox state in regulation of butane monooxygenase in Pseudomonas butanovora

J Bacteriol. 2008 Apr;190(8):2933-8. doi: 10.1128/JB.01409-07. Epub 2008 Feb 15.

Abstract

Pseudomonas butanovora possesses an alcohol-inducible alkane monooxygenase, butane monooxygenase (BMO), that initiates growth on C(2)-C(9) alkanes. A lacZ transcriptional reporter strain, P. butanovora bmoX::lacZ, in which the BMO promoter controls the expression of beta-galactosidase activity, was used to show that 1-butanol induced the BMO promoter in the presence or absence of O(2) when lactate-grown, BMO-repressed cells were washed free of lactate and incubated in NH(4)Cl-KNa phosphate buffer. In contrast, when lactate-grown cells of the reporter strain were incubated in phosphate buffer containing the mineral salts of standard growth medium, 1-butanol-dependent induction was significantly repressed at low O(2) (1 to 2% [vol/vol]) and totally repressed under anoxic conditions. The repressive effect of the mineral salts was traced to its copper content. In cells exposed to 1% (vol/vol) O(2), CuSO(4) (0.5 microM) repressed 1-butanol-dependent induction of beta-galactosidase activity. Under oxic conditions (20% O(2) [vol/vol]), significantly higher concentrations of CuSO(4) (2 microM) were required for almost complete repression of induction in lactate-grown cells. A combination of the Cu(2+) reducing agent Na ascorbate (100 microM) and CuSO(4) (0.5 microM) repressed the induction of beta-galactosidase activity under oxic conditions to the same extent that 0.5 microM CuSO(4) alone repressed it under anoxic conditions. Under oxic conditions, 2 microM CuSO(4) repressed induction of the BMO promoter less effectively in butyrate-grown cells of the bmoX::lacZ strain and of an R8-bmoX::lacZ mutant reporter strain with a putative BMO regulator, BmoR, inactivated. Under anoxic conditions, CuSO(4) repression remained highly effective, regardless of the growth substrate, in both BmoR-positive and -negative reporter strains.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Artificial Gene Fusion
  • Butanes / metabolism*
  • Copper / metabolism*
  • Culture Media / chemistry
  • Cytochrome P-450 CYP4A / biosynthesis*
  • Gene Expression Regulation, Bacterial*
  • Genes, Reporter
  • Lactic Acid / metabolism
  • Oxidation-Reduction
  • Oxygen / metabolism
  • Propionates / metabolism
  • Pseudomonas / enzymology*
  • Pseudomonas / physiology*
  • beta-Galactosidase / biosynthesis

Substances

  • Butanes
  • Culture Media
  • Propionates
  • Lactic Acid
  • butane
  • Copper
  • Cytochrome P-450 CYP4A
  • beta-Galactosidase
  • propionic acid
  • Oxygen