Testosterone 15beta-hydroxylation by solvent tolerant Pseudomonas putida S12

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 prote...

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Veröffentlicht in:	Journal of biotechnology 2007-08, Vol.131 (2), p.205
Hauptverfasser:	Ruijssenaars, Harald J, Sperling, Eric M G M, Wiegerinck, Peter H G, Brands, Ferry T L, Wery, Jan, de Bont, Jan A M
Format:	Artikel
Sprache:	eng
Schlagworte:	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
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Zusammenfassung:	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.
ISSN:	0168-1656