Conversion of Lactobacillus pentosus D-lactate dehydrogenase to a D-hydroxyisocaproate dehydrogenase through a single amino acid replacement.
The single amino acid replacement of Tyr52 with Leu drastically increased the activity of Lactobacillus pentosus NAD-dependent D-lactate dehydrogenase toward larger aliphatic or aromatic 2-ketoacid substrates by 3 or 4 orders of magnitude and decreased the activity toward pyruvate by about 30-fold, converting the enzyme into a highly active D-2-hydroxyisocaproate dehydrogenase.
Tokuda
Chizuka
C
Department of Applied Biological Science, Faculty of Science and Technology, Science University of Tokyo, Noda, Chiba 278-8510, Japan.
Ishikura
Yoshiro
Y
Shigematsu
Mayu
M
Mutoh
Hiroyuki
H
Tsuzuki
Shino
S
Nakahira
Yusaku
Y
Tamura
Yusuke
Y
Shinoda
Takeshi
T
Arai
Kazuhito
K
Takahashi
O
O
Taguchi
Hayao
H
D000429:Alcohol Oxidoreductases / Q000235:genetics*; D019943:Amino Acid Substitution; D001665:Binding Sites; D002208:Caproates / Q000737:chemistry / Q000378:metabolism*; D005818:Genetic Engineering; D007770:L-Lactate Dehydrogenase / Q000235:genetics*; D007778:Lactobacillus / Q000201:enzymology* / Q000235:genetics; D008958:Models, Molecular; D013379:Substrate Specificity
D002208:Caproates; C034527:isocaproic acid; D000429:Alcohol Oxidoreductases; D007770:L-Lactate Dehydrogenase; C021379:D-2-hydroxyacid dehydrogenase