Widely applicable background depletion step enables transaminase evolution through solid-phase screening

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 > Departement of Chemistry and Biochemistry

UniBE Contributor:

Planchestainer, Matteo and 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:

24 Oct 2019 00:05

Publisher DOI:

10.1039/C8SC05712E

BORIS DOI:

10.7892/boris.131231

URI:

https://boris.unibe.ch/id/eprint/131231

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