Molecular cloning and characterization of a novel glucansucrase from Leuconostoc mesenteroides subsp. mesenteroides LM34

• Published in: Biotechnology and bioprocess engineering Vol. 19; no. 4; pp. 605 - 612
• Main Authors: Kang, Hee-Kyoung, Nguyen, Thi Thanh Hanh, Jeong, Ha-Na, Park, Min-Eon, Kim, Doman
• Format: Journal Article
• Language: English
• Published: Heidelberg The Korean Society for Biotechnology and Bioengineering 01.07.2014; Springer Nature B.V; 한국생물공학회
• Subjects: Analysis; Bacteria; Biotechnology; Chemistry; Chemistry and Materials Science; Cloning; Dairy products; Enzymes; Escherichia coli; Fermentation; Food; Food processing industry; Genetic recombination; Gram-positive bacteria; Industrial and Production Engineering; Lactobacillus lactis; Lactose; Leuconostoc mesenteroides; Milk; Phenols; Polymerase chain reaction; Research Paper; Solidification; Studies; Sucrose; Sulfuric acid; Yogurt; 생물공학; kimchi; dextran; glucansucrase
• ISSN: 1226-8372; 1976-3816
• DOI: 10.1007/s12257-014-0116-3

Leuconostoc mesenteroides LM34 was isolated from kimchi, a traditional fermented Korean food. L. mesenteroides LM34 produced extracellular glucansucrase (DSRLM34), which is responsible for the synthesis of soluble glucan using sucrose. The DSRLM34 gene consists of a 4,503 bp open reading frame (ORF) and encodes an enzyme of 1,500 amino acids with an apparent molecular mass of 165 kDa. The deduced amino-acid sequence showed the highest amino-acid sequence identity (98%) to that of glucansucrase of Lactobacillus lactis . The gene was over-expressed in Escherichia coli strain and the recombinant enzyme (rDSRLM34) was purified. Both DSRLM34 and rDSRLM34 synthesized glucan mainly containing α-1, 6 glucosidic linkage and branched α-1, 3 glucosidic linkages. The enzyme exhibited optimum activity at 30°C and pH 5.0. DSRLM34 has promising potential as a thickening agent in sucrose-supplemented milk.