
Cloning and sequence analysis of the gene encoding Lactococcus lactis malolactic enzyme: Relationships with malic enzymes
Author(s) -
Denayrolles Muriel,
Aigle Michel,
LonvaudFunel Aline
Publication year - 1994
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1994.tb06679.x
Subject(s) - lactococcus lactis , malolactic fermentation , biology , malic enzyme , gene , peptide sequence , biochemistry , nucleic acid sequence , escherichia coli , microbiology and biotechnology , lactococcus , enzyme , amino acid , genetics , bacteria , lactic acid , dehydrogenase
Malolactic enzyme is the key enzyme in the degradation of L‐malic acid by lactic acid bacteria. Using degenerated primers designed from the first 20 N‐terminal amino acid sequence of lactococcal malolactic enzyme, a 60‐bp DNA fragment containing part of the mleS gene was amplified from Lactococcus lactis in a polymerase chain reaction. This specific probe was used to isolate two contiguous fragments covering the gene as a whole. The 1.9‐kb region sequenced contains an open reading frame of 1623 bp, coding a putative protein of 540 amino acids. The deduced amino acid sequence reveals that lactococcal putative protein (Mlep) is highly homologous to the malic enzyme of other organisms. Expression of the mleS gene in Escherichia coli results in malolactic activity.