Identification, expression and characterization of an R-ω-transaminase from Capronia semiimmersa

Applied Microbiology and Biotechnology
César IglesiasSonia Rodriguez Giordano

Abstract

Chiral amines are essential precursors in the production of biologically active compounds, including several important drugs. Among the biocatalytic strategies that have been developed for their synthesis, the use of ω-transaminases (ω-TA) appears as an attractive alternative allowing the stereoselective amination of prochiral ketones. However, the problems associated with narrow substrate specificity, unfavourable reaction equilibrium and expensive amine donors still hamper its industrial application. The search for novel enzymes from nature can contribute to expand the catalytic repertoire of ω-TA and help to circumvent some of these problems. A genome mining approach, based on the work described by Höhne et al., was applied for selection of potential R-ω-TA. Additional criteria were used to select an enzyme that differs from previously described ones. A candidate R-ω-TA from Capronia semiimmersa was selected, cloned and expressed in Escherichia coli. Interestingly, alignment of this enzyme with previously reported TA sequences revealed the presence of two additional amino acid residues in a loop close to the active site. The impact of this change was analysed with a structural model based on crystallized R-ω-TAs. Analysis of...Continue Reading

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Citations

Nov 6, 2018·Organic & Biomolecular Chemistry·Panagiotis Kelefiotis-StratidakisIoannis V Pavlidis
Nov 11, 2018·Applied and Environmental Microbiology·Cristina CoscolínManuel Ferrer
Dec 20, 2017·World Journal of Microbiology & Biotechnology·Erica E Ferrandi, Daniela Monti
Jan 29, 2020·Applied Microbiology and Biotechnology·Ekaterina Yu BezsudnovaKonstantin M Boyko
Apr 18, 2020·Applied Microbiology and Biotechnology·Stephen A KellyBrendan F Gilmore
Nov 27, 2020·Chembiochem : a European Journal of Chemical Biology·Aline TelzerowKerstin Steiner
Aug 5, 2021·Applied Biochemistry and Biotechnology·Dong-Xu JiaYu-Guo Zheng

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Methods Mentioned

BETA
electrophoresis

Software Mentioned

TA
YASARA
BLAST
Pred
GeneScript
R
PSI
Clustal Omega
Cap
MEGA7 Molecular Evolutionary Genetics Analysis

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