title: Testosterone 15beta-hydroxylation by solvent tolerant Pseudomonas putida S12.
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.
authors.affiliation: TNO Quality of Life, Julianalaan 67, 2628BC Delft, The Netherlands. harald.ruijssenaars@tno.nl
authors.forename: Harald J
authors.initials: HJ
authors.lastname: Ruijssenaars
authors.forename: Eric M G M
authors.initials: EM
authors.lastname: Sperling
authors.forename: Peter H G
authors.initials: PH
authors.lastname: Wiegerinck
authors.forename: Ferry T L
authors.initials: FT
authors.lastname: Brands
authors.forename: Jan
authors.initials: J
authors.lastname: Wery
authors.forename: Jan A M
authors.initials: JA
authors.lastname: de Bont
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D006900:Hydroxylation; D008954:Models, Biological; D016297:Mutagenesis, Site-Directed; D030781:Organisms, Genetically Modified; D016958:Pseudomonas putida / Q000235:genetics*; D012997:Solvents / Q000494:pharmacology*; D013379:Substrate Specificity; D013739:Testosterone / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D012997:Solvents; D013739:Testosterone; D003577:Cytochrome P-450 Enzyme System; C517094:15beta-hydroxylase CYP106A2, Bacillus megaterium