6GWI

The crystal structure of Halomonas elongata amino-transferase

Summary for 6GWI

• Entry DOI: 10.2210/pdb6gwi/pdb
• Descriptor: Putrescine aminotransferase, PYRIDOXAL-5'-PHOSPHATE, 1,2-ETHANEDIOL, ... (5 entities in total)
• Functional Keywords: amino-transferase, transferase
• Biological source: Halomonas elongata DSM 2581
• Total number of polymer chains: 2
• Total formula weight: 101664.30

Authors

Gourlay, L.J. (deposition date: 2018-06-25, release date: 2019-05-08, Last modification date: 2025-04-09)

Primary citation

Planchestainer, M.,Hegarty, E.,Heckmann, C.M.,Gourlay, L.J.,Paradisi, F.
Widely applicable background depletion step enables transaminase evolution through solid-phase screening.
Chem Sci, 10:5952-5958, 2019

PubMed Abstract: 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 (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.

PubMed: 31360401
DOI: 10.1039/c8sc05712e

Experimental method

X-RAY DIFFRACTION (2 Å)