Testosterone 15beta-hydroxylation by Solvent Tolerant Pseudomonas Putida S12

J Biotechnol. 2007 Aug 31;131(2):205-8. doi: 10.1016/j.jbiotec.2007.06.007. Epub 2007 Jun 23.

Abstract

A steroid 15beta-hydroxylating whole-cell solvent tolerant biocatalyst was constructed by expressing the Bacillus megaterium steroid hydroxylase CYP106A2 in the solvent tolerant Pseudomonas putida S12. Testosterone hydroxylation was improved by a factor 16 by co-expressing Fer, a putative Fe-S protein from Bacillus subtilis. In addition, the specificity for 15beta-hydroxylation was improved by mutating threonine residue 248 of CYP106A2 into valine. These new insights provide the basis for an optimized whole-cell steroid-hydroxylating biocatalyst that can be applied with an organic solvent phase.

Publication types

  • Evaluation Study

MeSH terms

  • Bacterial Proteins / genetics*
  • Bacterial Proteins / metabolism*
  • Cytochrome P-450 Enzyme System / genetics*
  • Cytochrome P-450 Enzyme System / metabolism*
  • Gene Expression Regulation, Bacterial
  • Hydroxylation
  • Models, Biological
  • Mutagenesis, Site-Directed
  • Organisms, Genetically Modified
  • Pseudomonas putida / genetics*
  • Solvents / pharmacology*
  • Substrate Specificity
  • Testosterone / metabolism*

Substances

  • Bacterial Proteins
  • Solvents
  • Testosterone
  • Cytochrome P-450 Enzyme System
  • 15beta-hydroxylase CYP106A2, Bacillus megaterium