title: Engineering of highly selective variants of Parvibaculum lavamentivorans alcohol dehydrogenase.
abstract: We present the development of highly selective variants of the Parvibaculum lavamentivorans alcohol dehydrogenase. Four amino acids (A158, N162, K202, L224) in the second sphere of the catalytic site were identified to determine the selectivity for 3-quinuclidone reduction significantly. The best variant (A158H/N162G/K202Q/L224W) was able to increase the ee for (R)-3-quinuclidinol production from 84.3 % (wild-type) to ≥99 % and concomitantly to enhance conversion by 43.5 %.
authors.affiliation: evocatal GmbH, Alfred-Nobel-Strasse 10, 40789 Monheim am Rhein (Germany).
authors.forename: Dominik
authors.initials: D
authors.lastname: Spickermann
authors.forename: Sascha
authors.initials: S
authors.lastname: Hausmann
authors.forename: Christian
authors.initials: C
authors.lastname: Degering
authors.forename: Ulrich
authors.initials: U
authors.lastname: Schwaneberg
authors.forename: Christian
authors.initials: C
authors.lastname: Leggewie
mesh_terms: D000426:Alcohol Dehydrogenase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D020561:Alphaproteobacteria / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D020134:Catalytic Domain; D008958:Models, Molecular; D009154:Mutation; D010084:Oxidation-Reduction; D015202:Protein Engineering; D011812:Quinuclidines / Q000378:metabolism*
chemical_list: D011812:Quinuclidines; C474540:3-quinuclidinol; D000426:Alcohol Dehydrogenase
keywords: biocatalysis; high-throughput screening; oxidoreductases; protein engineering; selectivity