Cloning, nucleotide sequence determination and expression of the genes encoding cytochrome P-450soy (soyC) and ferredoxinsoy (soyB) from Streptomyces griseus

Mol Microbiol. 1992 Aug;6(15):2125-34. doi: 10.1111/j.1365-2958.1992.tb01386.x.

Abstract

Xenobiotic transformation by Streptomyces griseus (ATCC13273) is catalysed by a cytochrome P-450, designated cytochrome P-450soy. A DNA segment carrying the structural gene encoding P-450soy (soyC) was cloned using an oligonucleotide probe constructed from the protein sequence of a tryptic peptide. Following DNA sequencing the deduced amino acid sequence of P-450soy was compared with that for P-450cam, revealing conservation of important structural components including the haem pocket. Expression of the cloned soyC gene product was demonstrated in Streptomyces lividans by reduced CO:difference spectral analysis and Western blotting. Downstream of soyC, a gene encoding a putative [3Fe-4S] ferredoxin (soyB), named ferredoxinsoy, was identified.

Publication types

  • Comparative Study

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Biodegradation, Environmental
  • Cytochrome P-450 Enzyme System / genetics*
  • DNA, Bacterial / isolation & purification
  • Ferredoxins / biosynthesis
  • Ferredoxins / genetics*
  • Gene Expression Regulation, Bacterial*
  • Genes, Bacterial*
  • Genetic Vectors
  • Glycine max / enzymology
  • Glycine max / genetics*
  • Molecular Sequence Data
  • Oligonucleotide Probes
  • Peptide Fragments / isolation & purification
  • Plant Proteins, Dietary / genetics
  • Sequence Homology
  • Streptomyces griseus / enzymology
  • Streptomyces griseus / genetics*

Substances

  • DNA, Bacterial
  • Ferredoxins
  • Oligonucleotide Probes
  • Peptide Fragments
  • Plant Proteins, Dietary
  • Cytochrome P-450 Enzyme System

Associated data

  • GENBANK/X63601