Cloning and function analysis of L-lactate dehydrogenase gene from Lactobacillus sp. MD-1.
- Author:
Jian LI
1
;
Yun TANG
;
Feng-Lai LIANG
;
Xin-Ping ZHANG
;
Ru-Lin LIU
Author Information
1. Life College, Nankai University, Tianjin 300071, China.
- Publication Type:Journal Article
- MeSH:
Amino Acid Sequence;
Base Sequence;
Cloning, Molecular;
L-Lactate Dehydrogenase;
chemistry;
genetics;
physiology;
Lactobacillus;
genetics;
Molecular Sequence Data
- From:
Chinese Journal of Biotechnology
2004;20(5):725-729
- CountryChina
- Language:Chinese
-
Abstract:
It was constructed that a genomic DNA library from Lactobacillus sp. MD-1 yielding D, L-lactic acid. The gene encoding L-lactate dehydrogenase (L-LDH) was cloned from the genomic library of strain MD-1 by complementation in E. coli FMJ144 which was lactate dehydrogenase and pyruvate-formate lyase double defective mutant. The nucleotide sequence of the ldhL gene predicted a protein of 316 amino acid starting with ATG. The putative molecular weight of the L-LDH amino acid sequence was 33.84kD. A putative typical promoter (-35 and -10 boxes) had been observed in the 5' noncoding region. An rho-independent transcriptional terminator has been observed in the 3' noncoding region. Three highly conserved regions (Gly13 approximately Asp50, Asp73 approximately Ileul00 and Asn123 approximately Arg154) with several conserved residues had been identified. Gly13 approximately Asp50 was NADH-binding site domain. Asp73 approximately Ileu100 and Asn123 approximately Arg154 were reported to be the active site domains. The ldhL and the L-LDH of Lactobacillus sp. MD-1 showed the low identity and similarity with other Lactobacilli, and the highest percentage were 61.9% and 68.9% respectively. All the above indicated this gene is a novel ldhL.
