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
Biology
Biotechnology
CYP106A2
Fer
Pseudomonas putida
Solvent tolerance
Steroid synthesis
Whole cell biocatalyst
Biocatalysts
Enzyme kinetics
Hydroxylation
Optimization
Solvents
Pseudomonas putida
Solvent tolerance
Steroid synthesis
Whole cell biocatalysts
Drug products
15 beta hydroxylase
bacterial protein
cytochrome P450
organic solvent
steroid monooxygenase
testosterone
threonine
valine
15beta hydroxylase CYP106A2, Bacillus megaterium
15beta-hydroxylase CYP106A2, Bacillus megaterium
solvent
article
Bacillus subtilis
biocatalyst
biotransformation
chemical structure
controlled study
high performance liquid chromatography
nonhuman
nuclear magnetic resonance
priority journal
protein expression
Pseudomonas putida
steroid hydroxylation
steroidogenesis
biological model
enzyme specificity
evaluation
gene expression regulation
genetics
hydroxylation
metabolism
site directed mutagenesis
transgenic organism
Bacillus megaterium
Bacillus subtilis
Pseudomonas putida
Bacterial Proteins
Cytochrome P-450 Enzyme System
Gene Expression Regulation, Bacterial
Hydroxylation
Models, Biological
Mutagenesis, Site-Directed
Organisms, Genetically Modified
Pseudomonas putida
Solvents
Substrate Specificity
Testosterone
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