Molecular cloning and characterization of a novel glucansucrase from Leuconostoc mesenteroides subsp. mesenteroides LM34
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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.
KeywordsLeuconostoc mesenteroides glucansucrase dextran kimchi
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- 1.Reynold, J. A., J. D. Kastello, D. G. Harrington, C. L. Crabbs, C. J. Peters, J. V. Jemski, G. H. Scott, and N. R. Di Luzio (1980) Glucan-induced enhancement of host resistance to selected infectious diseases. Infect. Immun. 30: 51–57.Google Scholar
- 3.Rachini, A., D. Pietrella, P. Lup, A. Torosantucci, P. Chiani, C. Bromuro, C. Projetti, F. Bistoni, A. Casone, and A. Vecchiarelli (2007) An anti-β-glucan monoclonal antibody inhibits growth and capsule formation of Cryptococcus neoformans in vitro and exerts therapeutic, anticryptococcal activity in vitro. Infect. Immun. 75: 5085–5094.CrossRefGoogle Scholar
- 14.Tamime, A. Y. and V. M. E. Marshall (1977) Microbiology and technology of fermented milks. pp. 57–52. In: B. E. Law (ed.). Microbiology and biochemistry of cheese and fermented milks. 2nd ed. Blackie Academic and Professional, London, UK.Google Scholar
- 15.Tamime, A. Y. and R. K. Robinson (2007) Yogurt science and technology. Woodhead Publishing Ltd., Cambridge, UK.Google Scholar
- 18.Kim, J. M., H. N. Seo, T. S. Hwang, S. H. Lee, and D. H. Park (2008) Characterization of expolysaccharide (EPS) produced by Weissella hellencia SKkimchi3 isolated from Kimchi. J. Micronbiol. 46: 535–541.Google Scholar
- 35.Kang, H. K., Y. M. Kim, and D. Kim (2008) Functional, genetic, and bioinformatic characterization of dextransucrase (DSRBCB4) gene in Leuconostoc mesenteroides B-1299CB4. J. Microbiol. Biotechnol. 18: 1050–1058.Google Scholar
- 37.Park, M. R., H. J. Ryu, D. Kim, J. Y. Choe, and J. F. Robyt (2001) Characterization of Leuconostoc mesenteroides B-742CB dex-transucrase expressed in Escherichia coli. J. Microbiol. Biotechn. 11: 628–635.Google Scholar