Molecular Cloning and Characterization of a Novel Glucansucrase from Leuconostoc mesenteroides subsp. mesenteroides LM34

다국어 초록 (Multilingual Abstract)

Leuconostoc mesenteroides LM34 was isolatedfrom kimchi, a traditional fermented Korean food.
L. mesenteroides LM34 produced extracellular glucansucrase(DSRLM34), which is responsible for the synthesis ofsoluble glucan using sucrose. The DSRLM34 gene consistsof a 4,503 bp open reading frame (ORF) and encodes anenzyme of 1,500 amino acids with an apparent molecularmass of 165 kDa. The deduced amino-acid sequenceshowed the highest amino-acid sequence identity (98%) tothat of glucansucrase of Lactobacillus lactis. The gene wasover-expressed in Escherichia coli strain and the recombinantenzyme (rDSRLM34) was purified. Both DSRLM34 andrDSRLM34 synthesized glucan mainly containing α-1, 6glucosidic linkage and branched α-1, 3 glucosidic linkages.
The enzyme exhibited optimum activity at 30oC andpH 5.0. DSRLM34 has promising potential as a thickeningagent in sucrose-supplemented milk.

참고문헌 (Reference)

1 Tamime, A. Y., "Yogurt science and technology" Woodhead Publishing Ltd. 2007

2 Broadbent, J. R., "Use of exopolysaccharide-producing cultures to improve the functionality of low fat cheese" 11 : 433-439, 2001

3 Meulenbeld, G. H., "Transglycosylation by Streptococcus mutans GS-5 glucosyltransferase-D: Acceptor specificity and engineering of reaction conditions" 70 : 363-369, 2000

4 Chang, Y. H., "Sporo-lactobacillus vineae sp. nov., a spore-forming lactic acid bacterium isolated from vineyard soil" 58 : 2316-2320, 2008

5 Mukerjea, R., "Simplified and improved methylation analysis of saccharides, using a modified procedure and thin-layer chromatography" 292 : 11-20, 1996

6 Eom, H. J., "Selection of psychrotrophic Leuconostoc spp. producing highly active dextransu-crase from lactate fermented vegetables" 117 : 61-67, 2007

7 Ruas-Madiedo, P., "Role of exopolysaccharides produced by Lactococcus lactis subsp. cremoris on the viscosity of fermented milks" 12 : 689-695, 2002

8 Kim, Y. M., "Purification and characterization of a novel glucansucrase from Leuconostoc lactis EG001" 165 : 384-391, 2010

9 Robyt, J. F., "Production, purification, and properties of dextransucrase from Leuconostoc mesenteroides NRRL B-512F" 68 : 95-111, 1979

10 Kim, D., "Production and selection of mutants of Leuconostoc mesenteroides constitutive for glucansu-crase" 16 : 659-664, 1994

11 Kang, H. K., "Molecular characterization and expression analysis of the glucansucrase DSRWC from Weissella cibaria synthesizing a α(1→6) glucan" 292 : 33-41, 2009

12 Tamime, A. Y., "Microbiology and biochemistry of cheese and fermented milks" Blackie Academic and Professional 57-52, 1977

13 Naessens, M., "Leuconostoc dextransucrase and dextran: Production, properties and applications" 80 : 845-860, 2005

14 Tanriseven, A., "Interpretation of dextran-sucrase inhibition at high sucrose concentrations" 245 : 97-104, 1993

15 Ayala-Hernandez, I., "Inter-actions between milk proteins and exopolysaccharides produced by Lactococcus lactis observed by scanning electron microscopy" 91 : 2583-2590, 2008

16 Remaud-Simeon, M., "Glucansucrases: Molecular engineering and oligosaccharide synthesis" 10 : 117-128, 2000

17 Reynold, J. A., "Glucan-induced enhancement of host resistance to selected infectious diseases" 30 : 51-57, 1980

18 강희경, "Functional, Genetic, and Bioinformatic Characterization of Dextransucrase(DSRBCB4) Gene in Leuconostoc mesenteroides B-1299CB4" 한국미생물·생명공학회 18 (18): 1050-1058, 2008

19 Shah, N. P., "Functional cultures and health benefits" 17 : 1262-1277, 2007

20 Shiby, V. K., "Fermented milks and milk products as functional foods" 53 : 482-496, 2013

21 Lacaze, G., "Emerging fermentation technologies: development of novel sourdoughs" 24 : 155-160, 2007

22 Kralj, S., "Efficient screening methods for glucosyltrans-ferase genes in Lactobacillus strains" 21 : 181-187, 2003

23 Singh, M., "Effect of purified oat â-glucan on fermentation of set-style yogurt mix" 77 : 195-201, 2012

24 Kang, H. K., "Directed evolution of a dextransucrase for increased constitutive activity and the synthesis of a highly branched dextran" 26 : 167-176, 2003

25 Robyt, J. F., "Dextransucrase and the mechanism for dextran biosynthesis" 343 : 3039-3048, 2008

26 Miller, A. W., "Detection of dextransucrase and levansucrase on polyacrylamide gels by the periodic acid-Schiff stain: Staining artifacts and their prevention" 156 : 357-363, 1986

27 Salminen, S., "Demonstration of safety of probiotics-a review" 44 : 93-106, 1998

28 Dubois, M., "Colorimetric method for determination of sugars and related substances" 28 : 350-356, 1956

29 Bejar, W., "Characterization of glucansucrase and dextran from Weis-sella sp. TN610 with potential as safe food additives" 52 : 125-132, 2013

30 Kim, J. M., "Characterization of expolysaccharide (EPS) produced by Weissella hellencia SKkimchi3 isolated from Kimchi" 46 : 535-541, 2008

31 Park, M. R., "Characterization of Leuconostoc mesenteroides B-742CB dextransucrase expressed in Escherichia coli" 11 : 628-635, 2001

32 Jeanes, A., "Characterization and classification of dextrans from ninety-six strains of bacteria" 76 : 5041-5052, 1954

33 Thompson, J. D., "CLUST-ALW: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, positions-specific gap penalties and weight matrix choice" 22 : 4673-4680, 1994

34 Majumder, A., "Application of response surface methodology for glucan production from Leu-conostoc dextranicum and its structural characterization" 75 : 150-156, 2009

35 Rachini, A., "An anti-β-glucan monoclonal antibody inhibits growth and capsule formation of Cryptococcus neoformans in vitro and exerts therapeutic, anticryptococcal activity in vitro" 75 : 5085-5094, 2007

36 Funame, K., "Aggregated form of dextransucrases from Leuconostoc mesenteroides NRRL B-512F and its constitutive mutant" 59 : 776-780, 1995

37 Bradford, M. M., "A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the prin-ciple of protein-dye binding" 72 : 248-254, 1976

38 Cote, G. L., "A method for surveying and classifying Leuconostoc spp. glucanases according to strain-dependent acceptor product patterns" 32 : 53-60, 2005

39 Laemmli, U. K., "(1970)Cleavage of structural proteins during the assembly of the head of bacteriophage T4" 227 : 680-685, 1970