Conversion of Lactobacillus pentosus D-lactate dehydrogenase to a D-hydroxyisocaproate dehydrogenase through a single amino acid replacement

Chizuka Tokuda; Yoshiro Ishikura; Mayu Shigematsu; Hiroyuki Mutoh; Shino Tsuzuki; Yusaku Nakahira; Yusuke Tamura; Takeshi Shinoda; Kazuhito Arai; O Takahashi; Hayao Taguchi

doi:10.1128/JB.185.16.5023-5026.2003

Conversion of Lactobacillus pentosus D-lactate dehydrogenase to a D-hydroxyisocaproate dehydrogenase through a single amino acid replacement

J Bacteriol. 2003 Aug;185(16):5023-6. doi: 10.1128/JB.185.16.5023-5026.2003.

Authors

Chizuka Tokuda¹, Yoshiro Ishikura, Mayu Shigematsu, Hiroyuki Mutoh, Shino Tsuzuki, Yusaku Nakahira, Yusuke Tamura, Takeshi Shinoda, Kazuhito Arai, O Takahashi, Hayao Taguchi

Affiliation

¹ Department of Applied Biological Science, Faculty of Science and Technology, Science University of Tokyo, Noda, Chiba 278-8510, Japan.

Abstract

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Alcohol Oxidoreductases / genetics*
Amino Acid Substitution*
Binding Sites
Caproates / chemistry
Caproates / metabolism*
Genetic Engineering
L-Lactate Dehydrogenase / genetics*
Lactobacillus / enzymology*
Lactobacillus / genetics
Models, Molecular
Substrate Specificity

Substances

Caproates
isocaproic acid
Alcohol Oxidoreductases
L-Lactate Dehydrogenase
D-2-hydroxyacid dehydrogenase