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

V. Dartois, V. Phalip, P. Schmitt, C. Diviès

Research output: Contribution to journalArticle

20 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)291-302
Number of pages12
JournalResearch in Microbiology
Volume146
Issue number4
DOIs
StatePublished - Jan 1 1995

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Biology

Keywords

  • Gene sequencing, Protein purification, Diacetyl production, L. mesenteroides subsp. cremoris, HicDH
  • LDH, Leuconostoc

Fingerprint Dive into the research topics of 'Purification, properties and DNA sequence of the D-lactate dehydrogenase from Leuconostoc mesenteroides subsp. cremoris'. Together they form a unique fingerprint.

  • Cite this