Purification, properties and DNA sequence of the D-lactate dehydrogenase from Leuconostoc mesenteroides subsp. cremoris

Res Microbiol. 1995 May;146(4):291-302. doi: 10.1016/0923-2508(96)81052-7.

Abstract

The complete sequence of the D-lactate dehydrogenase (D-ldh) gene from Leuconostoc mesenteroides subsp. cremoris, cloned in Escherichia coli, were determined. The deduced amino acid sequence showed homologies with all members of the D-specific-2-hydroxyacid dehydrogenase family. Furthermore, the essential residues detected so far as being involved in catalysis were also conserved. Purification of the enzyme revealed physico-chemical properties corresponding to those predicted from the sequence. The active enzyme was a dimer of 40-kDa subunits. The Km values for pyruvate, lactate, NADH and NAD were 0.3, 19, 0.03 and 0.16 mM, indicating that the enzyme reduced pyruvate in vivo. Besides the D-LDH activity, L. mesenteroides subsp. cremoris also displayed HicDH enzymatic activity, catalysing the reduction of pyruvate analogs. The purified D-LDH displayed low HicDH-type activity; therefore, differences in specificity profiles between the crude extract and the purified enzyme suggested the occurrence of a specific HicDH.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cloning, Molecular
  • DNA, Bacterial / chemistry*
  • Electrophoresis, Polyacrylamide Gel
  • In Vitro Techniques
  • L-Lactate Dehydrogenase / chemistry
  • L-Lactate Dehydrogenase / genetics*
  • L-Lactate Dehydrogenase / isolation & purification
  • Lactate Dehydrogenases*
  • Leuconostoc / enzymology*
  • Leuconostoc / genetics
  • Molecular Sequence Data
  • Polymerase Chain Reaction

Substances

  • DNA, Bacterial
  • Lactate Dehydrogenases
  • L-Lactate Dehydrogenase
  • D-lactate dehydrogenase

Associated data

  • GENBANK/L29327