Print Email Facebook Twitter Testosterone 15β-hydroxylation by solvent tolerant Pseudomonas putida S12 Title Testosterone 15β-hydroxylation by solvent tolerant Pseudomonas putida S12 Author Ruijssenaars, H.J. Sperling, E.M.G.M. Wiegerinck, P.H.G. Brands, F.T.L. Wery, J. de Bont, J.A.M. TNO Kwaliteit van Leven Publication year 2007 Abstract 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. © 2007 Elsevier B.V. All rights reserved. Subject BiologyBiotechnologyCYP106A2FerPseudomonas putidaSolvent toleranceSteroid synthesisWhole cell biocatalystBiocatalystsEnzyme kineticsHydroxylationOptimizationSolventsPseudomonas putidaSolvent toleranceSteroid synthesisWhole cell biocatalystsDrug products15 beta hydroxylasebacterial proteincytochrome P450organic solventsteroid monooxygenasetestosteronethreoninevaline15beta hydroxylase CYP106A2, Bacillus megaterium15beta-hydroxylase CYP106A2, Bacillus megateriumsolventarticleBacillus subtilisbiocatalystbiotransformationchemical structurecontrolled studyhigh performance liquid chromatographynonhumannuclear magnetic resonancepriority journalprotein expressionPseudomonas putidasteroid hydroxylationsteroidogenesisbiological modelenzyme specificityevaluationgene expression regulationgeneticshydroxylationmetabolismsite directed mutagenesistransgenic organismBacillus megateriumBacillus subtilisPseudomonas putidaBacterial ProteinsCytochrome P-450 Enzyme SystemGene Expression Regulation, BacterialHydroxylationModels, BiologicalMutagenesis, Site-DirectedOrganisms, Genetically ModifiedPseudomonas putidaSolventsSubstrate SpecificityTestosterone To reference this document use: http://resolver.tudelft.nl/uuid:12415d3a-8b19-4cb1-b9ac-b72097252d02 DOI https://doi.org/10.1016/j.jbiotec.2007.06.007 TNO identifier 240153 ISSN 0168-1656 Source Journal of Biotechnology, 131 (2), 205-208 Document type article Files To receive the publication files, please send an e-mail request to TNO Library.