Planchestainer, Matteo; Hegarty, Eimear; Heckmann, Christian M.; Gourlay, Louise J.; Paradisi, Francesca (2019). Widely applicable background depletion step enables transaminase evolution through solid-phase screening. Chemical Science, 10(23), pp. 5952-5958. The Royal Society of Chemistry 10.1039/C8SC05712E
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Directed evolution of transaminases is a widespread technique in the development of highly sought-after biocatalysts for industrial applications. This process, however, is challenged by the limited availability of effective high-throughput protocols to evaluate mutant libraries. Here we report a rapid, reliable, and widely applicable background depletion method for solid-phase screening of transaminase variants, which was successfully applied to a transaminase from Halomonas elongata (HEWT), evolved through rounds of random mutagenesis towards a series of diverse prochiral ketones. This approach enabled the identification of transaminase variants in viable cells with significantly improved activity towards para-substituted acetophenones (up to 60-fold), as well as tetrahydrothiophen-3-one and related substrates. Rationalisation of the mutants was assisted by determination of the high-resolution wild-type HEWT crystal structure presented herein.
Item Type: |
Journal Article (Original Article) |
|---|---|
Division/Institute: |
08 Faculty of Science > Department of Chemistry, Biochemistry and Pharmaceutical Sciences (DCBP) |
UniBE Contributor: |
Planchestainer, Matteo, Paradisi, Francesca |
Subjects: |
500 Science > 570 Life sciences; biology 500 Science > 540 Chemistry |
ISSN: |
2041-6520 |
Publisher: |
The Royal Society of Chemistry |
Funders: |
[UNSPECIFIED] Biotechnology and Biological Sciences Research Council |
Projects: |
[UNSPECIFIED] BB/P002536/1
[UNSPECIFIED] BB/M008770 |
Language: |
English |
Submitter: |
Francesca Paradisi |
Date Deposited: |
09 Jul 2019 09:45 |
Last Modified: |
05 Dec 2022 15:28 |
Publisher DOI: |
10.1039/C8SC05712E |
BORIS DOI: |
10.7892/boris.131231 |
URI: |
https://boris.unibe.ch/id/eprint/131231 |
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