KSBB Journal

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
권호 표지
DOI QR코드

DOI QR Code

Isolation of a Marine-derived Flammeovirga sp. mbrc-1 Strain and Characterization of Its Agarase

해양성 Flammeovirga sp. mbrc-1 균주의 분리 및 한천분해기능의 특성조사

  • Jang, Hye-Ji (Department of Pharmaceutical Engineering, Silla University) ;
  • Lee, Dong-Geun (Department of Pharmaceutical Engineering, Silla University) ;
  • Lee, Seung-Woo (Department of Pharmaceutical Engineering, Silla University) ;
  • Jeon, Myong-Je (Department of Pharmaceutical Engineering, Silla University) ;
  • Chun, Won-Ju (Department of Pharmaceutical Engineering, Silla University) ;
  • Kwon, Kae-Kyoung (Marine Biotechnology Research Center, KORDI) ;
  • Lee, Hee-Soon (Marine Biotechnology Research Center, KORDI) ;
  • Lee, Sang-Hyeon (Department of Pharmaceutical Engineering, Silla University)
  • 장혜지 (신라대학교 의생명과학대학 제약공학과) ;
  • 이동근 (신라대학교 의생명과학대학 제약공학과) ;
  • 이승우 (신라대학교 의생명과학대학 제약공학과) ;
  • 전명제 (신라대학교 의생명과학대학 제약공학과) ;
  • 천원주 (신라대학교 의생명과학대학 제약공학과) ;
  • 권개경 (한국해양연구원) ;
  • 이희순 (한국해양연구원) ;
  • 이상현 (신라대학교 의생명과학대학 제약공학과)
  • Received : 2011.10.28
  • Accepted : 2011.11.29
  • Published : 2011.12.31
Download PDF
⟨ Previous Next ⟩

Abstract

A novel agar-degrading bacterium mbrc-1 was isolated from seashore of Kyungpo at Gangwon province and cultured in marine broth 2216 medium. Isolated bacterium mbrc-1 was named as Flammeovirga sp. mbrc-1 based on the 16S rDNA sequence. Its agarase showed maximum activity of 923 units/L at pH 7.0 and $45^{\circ}C$ and sustained 90% remaining activity after exposed to $45^{\circ}C$ for 2 hours. The enzyme hydrolyzed agarose to yield neoagarohexaose (18.5%), neoagarotetraose (38%) and neoagarobiose (43.5%), indicating that the enzyme is ${\beta}$-agarase. Thus, isolated bacterium and its ${\beta}$-agarase would be useful for the industrial production of neoagarotetraose and neoagarobiose.

Keywords

References

  1. Duckworth, M. and W. Yaphe (1971) Structure of agar. I. Fractionation of a complex mixture of polysaccharides. Carbohyd. Res. 16: 189-197. https://doi.org/10.1016/S0008-6215(00)86113-3
  2. Hong, J.-H., J.-J. Lee, S.-H. Hur, H. Choi, and J.-Y. Kong (2001) Effect of agarooligosaccharides on the growth of intestinal bacteria. J. Food Hyg. Safety 16: 11-15.
  3. Do, J.-H. (1997) Extraction and purification of agar from Gelidium amansii. J. Kor. Fish. Soc. 30: 423-427.
  4. Yang, J.-I., L.-C. Chen, Y.-Y. Shih, C. Hsieh, C.-Y. Chen, W.-M. Chen, and C.-C. Chen (2011) Cloning and characterization of ${\beta}$-agarase AgaYT from Flammeovirga yaeyamensis strain YT. J. Biosci. Bioeng. 112: 225-232. https://doi.org/10.1016/j.jbiosc.2011.05.016
  5. Potin, P., C. Richard, C. Rochas, and B. Kliarg (1993) Purification and characterization of the ${\alpha}$-agarase from Alteromonas agarlyticus (Cataldi) comb. nov., strain GJ1B. Eur. J. Biochem. 214: 599-607. https://doi.org/10.1111/j.1432-1033.1993.tb17959.x
  6. Kirimura, K., N. Masuda, Y. Iwasaki, H. Nakagawa, R. Kobayashi, and S. Usami (1999) Purification and characterization of a novel ${\beta}$-agarase from an alkalophilic bacterium, Alteromonas sp. E-1. J. Biosci. Bioeng. 87: 436-441. https://doi.org/10.1016/S1389-1723(99)80091-7
  7. Schafer, H. (2007) Isolation of Methylophaga spp. from marine dimethylsulfide-degrading enrichment cultures and identification of polypeptides induced during growth on dimethylsulfide. Appl. Environ. Microbiol. 73: 2580-2591. https://doi.org/10.1128/AEM.02074-06
  8. Suzuki, H., Y. Sawai, T. Suzuki, and K. Kawai (2002) Purification and characterization of an extracellular ${\alpha}$neoagarooligosaccharide hydrolase from Bacillus sp. MK03. J. Biosci. Bioeng. 93: 456-463. https://doi.org/10.1016/S1389-1723(02)80092-5
  9. 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. https://doi.org/10.5352/JLS.2005.15.6.884
  10. Lee, D.-G., N. Y. Kim, M. K. Jang, and S.-H. Lee (2007) Isolation and characterization of a marine bacterium Thalassomonas sp. SL-5 producing ${\beta}$-agarase. J. Life Sci. 17: 70-75. https://doi.org/10.5352/JLS.2007.17.1.070
  11. Lee, D.-G., O.-H. Lee, H. J. Jang, M.-K. Jang, K. H. Yoo, and S.-H. Lee (2008) Isolation and characterization of a marine derived bacterium Glaciecola sp. SL-12 producing ${\beta}$-agarase. J. Life Sci. 18: 58-62. https://doi.org/10.5352/JLS.2008.18.1.058
  12. Jang, M.-K., O. H. Lee, K. H. Yoo, D.-G. Lee, and S.-H. Lee (2007) Secretory overexpression of ${\beta}$-agarase in Bacillus subtilis and antibacterial activity of enzymatic products. J. Life Sci. 17: 1601-1604. https://doi.org/10.5352/JLS.2007.17.11.1601
  13. 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): structure-function relationships and improved solubility. Biosci. Biotechnol. Biochem. 59: 1933-1937. https://doi.org/10.1271/bbb.59.1933
  14. 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, J.-H. Lee, S.-J. Kim, and S.-H. Lee (2006) Cloning, expression, and characterization of a glycoside hydrolase family 50 ${\beta}$-agarase from marine Agarivorans isolate. Biotechnol. Lett. 28: 1925-1932. https://doi.org/10.1007/s10529-006-9171-y
  15. Kobayashi, R., M. Takisada, T. Suzuki, K. Kirimura, and S. Usami (1997) Neoagarobiose as a novel moisturizer with whitening effect. Biosci. Biotechnol. Biochem. 61: 162-163. https://doi.org/10.1271/bbb.61.162
  16. Jang, M.-K., D.-G. Lee, N.-Y. Kim, K.-H. Yu, H. J. Jang, S. W. Lee, H. J. Jang, Y. J. Lee, and S.-H. Lee (2009) Purification and characterization of neoagarotetraose from hydrolyzed agar. J. Microbiol. Biotechnol. 19: 1197-1200.
  17. Duckworth, M. and W. Yaphe (1970) Thin-layer chromatographic analysis of enzymic hydrolysate of agar. J. Chromatogr. 49: 482-487. https://doi.org/10.1016/S0021-9673(00)93663-X
  18. 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. https://doi.org/10.1139/m77-100
  19. Shoichi, H. and Y. Akira (2007) Flammeovirga kamogawensis sp. nov., isolated from coastal seawater in Japan. Int. J. Syst. Evol. Microbiol. 57: 1327-1330. https://doi.org/10.1099/ijs.0.64977-0
  20. Takshashi, M., K. Suzuki, and Y. Nakagawa (2006) Emendation of the genus Flammeovirga and Flammeovirga aprica with the proposal of Flammeovirga arenaria nom. rev., comb. nov. and Flammeovirga yaeyamensis sp. nov. Int. J. Syst. Evol. Microbiol. 56: 2095-2100. https://doi.org/10.1099/ijs.0.64324-0
  21. Kim, B.-J., S.-H. Hwang, H.-J. Kim, Y.-S. Kang, S.-D. Ha, and J.-Y. Kong (1999) Characteristics of ${\beta}$-agarase produced by marine bacterium Bacillus cereus ASK202. Kor. J. Biotechnol. Bioeng. 14: 96-102.
  22. Jung, I. S., Y. J. Kim, H. J., Song, S. W. Gal, and Y. J. Choi (2008) Purification and properties of a novel extracellular agarase from marine bacterium, Sphingomonas paucimobilis AS-1. J. Life Sci. 18: 103-108. https://doi.org/10.5352/JLS.2008.18.1.103
  23. 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.
  24. Ohta, Y., Y. Hatada, M. Miyazaki, Y. Nogi, S. Ito, and K. Horikoshi (2005) Purification and characterization of a novel alpha-agarase from a Glaciecola sp. Curr. Microbiol. 50: 212-216. https://doi.org/10.1007/s00284-004-4435-z
  25. 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. Biotechnol. Biochem. 59: 1933-1939. https://doi.org/10.1271/bbb.59.1933
  26. Lee, S. A., J. U. Kim, J. G. Jung, I. H. Kim, S. H. Lee, S. J. Kim, and J. H. Lee (2006) Production of ${\beta}$-agarase in batch and fed-batch culture by Agarivorans sp. JA-1. Kor. J. Biotechnol. Bioeng. 21: 389-393.
  27. Kong, J. Y., S. K. Bae, S. H. Hwang, S. D. Ha, H. T. Kim, S. K. Kim, and B. J. Kim (1996) Purification of extracellular agarase from marine bacterium (Pseudomonas sp. W7) and molecular cloning of the agarase gene. Kor. J. Biotechnol. Bioeng. 11: 37-45.

Cited by

  1. Industrial Applications of Saccharification Technology for Red Seaweed Polysaccharide vol.29, pp.5, 2014, https://doi.org/10.7841/ksbbj.2014.29.5.307
  2. Characterization of α-agarase from Alteromonas sp. SH-1 vol.31, pp.2, 2016, https://doi.org/10.7841/ksbbj.2016.31.2.113