6gwi

Publication Abstract from PubMed

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.

Widely applicable background depletion step enables transaminase evolution through solid-phase screening.,Planchestainer M, Hegarty E, Heckmann CM, Gourlay LJ, Paradisi F Chem Sci. 2019 May 9;10(23):5952-5958. doi: 10.1039/c8sc05712e. eCollection 2019 , Jun 21. PMID:31360401^[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.