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논문검색

Cloning and expression of D(-)-lactate dehydrogenase (D-LDH) gene from Lactobacillus sp. RKY 2

초록

영어

Lactobacillus sp. RKY2, which was isolated from a Korean traditional fermented food soybean paste, previously proved to be an efficient lactic acid bacterium for the production of lactic acid, because it could produce a large amount of lactic acid, with high productivity. The structural gene of D(-)-lactate dehydrogenase (D-LDH) was cloned from Lactobacillus sp. RKY2. The gene from chromosomal DNA was amplified by polymerase chain reaction (PCR) using a pair of
conserved sequences existed in D-LDH as primers. Approximately 1 kb of PCR product in size was cloned into an gene expression vector and the nucleotide sequence was determined. Analysis of this sequence identified a putative 999 bp D-LDH open reading frame that encodes a polypeptide of 332 amino acid residues with a deduced molecular mass of 36.5 kDa. After IPTG induction, the D-LDH was expressed in Escherichia coli. The electrophoretically pure enzyme
preparation migrated as a single band corresponding to the deduced molecular mass during SDS-PAGE. The amount of lactic acid produced by D-LDH mutant and its ldh-complemented derivative was determined by HPLC and anzymatic analysis.

저자정보

  • Jae-Hoon Jeong School of Biological Sciences and Technology, Chonnam National University
  • Jin-Nam Kim School of Biological Sciences and Technology, Chonnam National University
  • Young-Jung Wee School of Biological Sciences and Technology, Chonnam National University
  • Hwa-Won Ryu School of Biological Sciences and Technology, Chonnam National University

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