title: The engineering of a more thermally stable lactate dehydrogenase by reduction of the area of a water-accessible hydrophobic surface.
abstract: A site-directed mutant of Bacillus stearothermophilus lactate dehydrogenase (lactate:NAD+ oxidoreductase, EC 1.1.1.27) has been engineered in which the conserved hydrophobic residue isoleucine-250 has been replaced by the more hydrophilic residue asparagine. This isoleucine forms a large part of a water-accessible, hydrophobic surface in the active site of the apo-enzyme which is covered by the B-face of the nicotinamide ring when coenzymes are bound. Reduction in the area of this hydrophobic surface results in the mutant tetramer being more thermally stable than the wild-type enzyme.
authors: Department of Biochemistry, University of Bristol, U.K.
authors: D B
authors: DB
authors: Wigley
authors: A R
authors: AR
authors: Clarke
authors: C R
authors: CR
authors: Dunn
authors: D A
authors: DA
authors: Barstow
authors: T
authors: T
authors: Atkinson
authors: W N
authors: WN
authors: Chia
authors: H
authors: H
authors: Muirhead
authors: J J
authors: JJ
authors: Holbrook
mesh_terms: D001665:Binding Sites; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D001411:Geobacillus stearothermophilus / Q000201:enzymology / Q000235:genetics; D007532:Isoleucine; D007770:L-Lactate Dehydrogenase; D009154:Mutation; D013696:Temperature
chemical_list: D007532:Isoleucine; D007770:L-Lactate Dehydrogenase