Testosterone 15β-hydroxylation by solvent tolerant Pseudomonas putida S12

A steroid 15β-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...

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Veröffentlicht in:	Journal of biotechnology 2007-08, Vol.131 (2), p.205-208
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:	Bacillus megaterium Bacillus subtilis Biological and medical sciences Biotechnology CYP106A2 Fer Fundamental and applied biological sciences. Psychology Pseudomonas putida Solvent tolerance Steroid synthesis Whole cell biocatalyst
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Beschreibung
Zusammenfassung:	A steroid 15β-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 15β-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 1873-4863
DOI:	10.1016/j.jbiotec.2007.06.007