Testosterone 15β-hydroxylation by solvent tolerant Pseudomonas putida S12

article





Ruijssenaars, H.J.

Sperling, E.M.G.M.

Wiegerinck, P.H.G.

Brands, F.T.L.

Wery, J.

Bont, J.A.M.de
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.
Topics































































BiotechnologyCYP106A2FerPseudomonas 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 megateriumSolventBacillus 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
TNO Identifier
240153
Repository link
https://resolver.tno.nl/uuid:12415d3a-8b19-4cb1-b9ac-b72097252d02
ISSN
01681656
Source
Journal of Biotechnology, 131(2), pp. 205-208.
Pages
205-208
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