title: Acceleration of an aldo-keto reductase by minimal loop engineering.
abstract: Aldo-keto reductases tighten coenzyme binding by forming a hydrogen bond across the pyrophosphate group of NAD(P)(H). Mutation of the hydrogen bonding anchor Lys24 in Candida tenuis xylose reductase prevents fastening of the "safety belt" around NAD(H). The loosened NAD(H) binding leads to increased turnover numbers (k(cat)) for reductions of bulky-bulky ketones at constant substrate and coenzyme affinities (i.e. K(m Ketone), K(m NADH)).
authors.affiliation: Institute of Biotechnology and Biochemical Engineering, Graz University of Technology, Member of NAWI Graz, Petersgasse 12, 8010 Graz, Austria.
authors.forename: C
authors.initials: C
authors.lastname: Krump
authors.affiliation: Institute of Organic Chemistry, University of Vienna, Währingerstraße 38, 1090 Vienna, Austria.
authors.forename: M
authors.initials: M
authors.lastname: Vogl
authors.affiliation: Institute of Organic Chemistry, University of Vienna, Währingerstraße 38, 1090 Vienna, Austria.
authors.forename: L
authors.initials: L
authors.lastname: Brecker
authors.affiliation: Institute of Biotechnology and Biochemical Engineering, Graz University of Technology, Member of NAWI Graz, Petersgasse 12, 8010 Graz, Austria Austrian Centre of Industrial Biotechnology, Petersgasse 14, 8010 Graz, Austria.
authors.forename: B
authors.initials: B
authors.lastname: Nidetzky
authors.affiliation: Institute of Biotechnology and Biochemical Engineering, Graz University of Technology, Member of NAWI Graz, Petersgasse 12, 8010 Graz, Austria Austrian Centre of Industrial Biotechnology, Petersgasse 14, 8010 Graz, Austria.
authors.forename: R
authors.initials: R
authors.lastname: Kratzer
mesh_terms: D000449:Aldehyde Reductase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D000074408:Aldo-Keto Reductases; D002175:Candida / Q000201:enzymology; D006860:Hydrogen Bonding; D007700:Kinetics; D008958:Models, Molecular; D009154:Mutation; D009243:NAD / Q000378:metabolism; D011487:Protein Conformation; D015202:Protein Engineering
chemical_list: D009243:NAD; D000074408:Aldo-Keto Reductases; D000449:Aldehyde Reductase
keywords: Aldo-keto reductase; chiral ethyl mandelates; enzyme engineering; saturation transfer difference NMR