Browse > Article
http://dx.doi.org/10.5352/JLS.2007.17.11.1601

Secretory Overexpression of β-Agarase in Bacillus subtilis and Antibacterial Activity of Enzymatic Products  

Jang, Min-Kyung (Department of Bioscience and Biotechnology, College of Engineering, Silla University)
Lee, Ok-Hee (Department of Bioscience and Biotechnology, College of Engineering, Silla University)
Yoo, Ki-Hwan (Department of Bioscience and Biotechnology, College of Engineering, Silla University)
Lee, Dong-Geun (Department of Bioscience and Biotechnology, College of Engineering, Silla University)
Lee, Sang-Hyeon (Department of Bioscience and Biotechnology, College of Engineering, Silla University)
Publication Information
Journal of Life Science / v.17, no.11, 2007 , pp. 1601-1604 More about this Journal
Abstract
The gene for ${\beta}-agarase$ of an Agarivorans sp. JA-1 was expressed in Bacillus subtilis DB104, 168 and ISW1214 strains for mass-production. Among 3 host strains, B. subtilis ISW1214 secreted the highest amount of recombinant ${\beta}-agarase$ with a specific activity of 201 U/mg and 360 mg of protein into culture broth. This was approximately 130-fold higher than the production in E. coli as an expression host. Recombinant enzyme produced neoagarooligosaccharides such as neoagarohexaose, neoagarotetraose, and neoagarobiose from agar. Produced neoagarooligosaccharides showed antibacterial activities against gram-negative E. coli and gram-positive B. subtilis at a concentration of 1.5%. These data suggest that neoagarooligosaccharides could be an useful preservative for food industry.
Keywords
${\beta}-agarase$; antibacterial activity; neoagarooligosaccharides; secretory expression;
Citations & Related Records
Times Cited By KSCI : 1  (Citation Analysis)
연도 인용수 순위
1 Somogy, M. 1952. Notes on sugar determination. J. Biol. Chem. 195, 19-23.
2 Yoshizawa, Y., A. Ametani, J. Tsunehiro, K. Nomura, M. Itoh, F. Fukui and S. Kaminogawa. 1995. Macrophage stimulation activity of the polysaccharide fraction from a marine alga (Porphyra yezoensis): structurefunction relationships and improved solubility. Biosci. Biotech. Bioch. 59, 1933-1939.   DOI   ScienceOn
3 Kim, K. H., J. Y. Kim, H. B. Kim and D. S. Lee. 2001. Expression of a $\beta$-1,3-glucanase gene from Bacillus circulans in B. subtilis and B. megaterium. Kor. J. Microbiol. 37, 253-258.
4 Kobayashi, R. Μ. Takisada, Τ. Suzuki, Κ. Kirimura and S. Usami. 1997. Neoagarobiose as a novel moisturizer with whitening effect. Biosci. Biotechnol. Biochem. 61, 62-63.
5 Kohno, T., H. Kitagawa, T. Hiraga. 1990. Production of hetero-oligosaccharides. pp. 87-105, In Kukami, G. K. (ed.), Shokuhin sangyo bioreactor system, Jissen bioreactor, Shokuhin Kagaku Shimbunsa, Tokyo.
6 Kono, T. and H. Hidaka. 1989. Properties and production of neoagarooligosaccharide. J. Agric. Chem. Soc. Jpn. 63, 1126-1129.
7 Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680-685.   DOI   ScienceOn
8 Lee, D. G., G. T. Park, N. Y. Kim, E. J. Lee, M. K. Jang, Y. G. Shin, G. S. Park, T. M. Kim, J. H. Lee, S. J. Kim and S. H. Lee. 2006. Cloning, expression, and characterization of a glycoside hydrolase family 50 beta-agarase from a marine Agarivorans isolate. Biotechnol. Lett. 28, 1925-1932.   DOI   ScienceOn
9 Ohta, Y., Y. Nogi, M. Miyazaki, Z. Li, Y. Hatada, S. Ito and Koki Horikishi. 2004. Enzymatic properties and nucleotide and amino acid sequences of a thermostable $\beta$- agarase from the novel marine isolate, JAMB-A94. Biosci. Biotech. Bioch. 68, 1073-1081.   DOI   ScienceOn
10 Park, G. T., D. G. Lee, N. Y. Kim, E. J. Lee, J. G. Jung, J. H. Lee, M. S. Heo and S. H. Lee. 2005. Isolation and characterization of a marine bacterium producing thermotolerant agarase. J. Life Sci. 15, 884-888.   과학기술학회마을   DOI   ScienceOn
11 Duckworth, M. and W. Yaphe. 1971. Structure of agar. I. Fractionation of a complex mixture of polysaccharides. Carbohydr. Res. 16, 189-197.   DOI   ScienceOn
12 Sambrook, J. and D. W. Russell. 1989. Molecular Cloning: A Laboratory Manual. 2nd eds., Cold Spring Harbor Laboratory Press, New York, New York.
13 Araki, T., Z. Lu and T. Morishita. 1998 Optimization of parameters for isolation of protoplasts from Gracilaria verrucosa (Rhodophyta). J. Mar. Biotechnol. 6, 193-197.
14 Do, J. H. 1997. Extraction and purification of agar from Gelidium amansii. J. Korean Fish. Soc. 30, 423-427.
15 Groleau, D. and W. Yaphe. 1977. Enzymatic hydrolysis of agar: purification and characterization of $\beta$-neoagarotetraose hydrolase from Peudomonas atlantica. Can. J. Microbiol. 23, 672-679.   DOI   ScienceOn
16 Kim, B. J., Η. J. Kim, S. D. Ha, S. H. Hwang, D. S. Byun, T. H. Lee and J. Y. Kong. 1999. Purification and characterization of $\beta$-agarase from marine bacterium Bacillus cereus ASK202. Biotechnol. Lett. 21, 1011-1105.   DOI   ScienceOn