Browse > Article
http://dx.doi.org/10.7841/ksbbj.2016.31.2.113

Characterization of α-agarase from Alteromonas sp. SH-1  

Lee, Sol-Ji (Department of Bioscience, Graduate School, Silla University)
Shin, Da-Young (Department of Pharmaceutical Engineering, Silla University)
Kim, Jae-Deog (Department of Green-Chemistry Convergence Engineering, Silla University)
Lee, Dong-Geun (Department of Bioscience, Graduate School, Silla University)
Lee, Sang-Hyeon (Department of Bioscience, Graduate School, Silla University)
Publication Information
KSBB Journal / v.31, no.2, 2016 , pp. 113-119 More about this Journal
Abstract
A novel agar-degrading marine bacterium, SH-1 strain, was isolated from seashore of Namhae at Gyeongnam province, Korea. The SH-1 strain exhibited 98% similarity with Alteromonas species based on 16S rDNA sequencing and named as Alteromonas sp. SH-1. Alteromonas sp. SH-1 showed agarase activity of 348.3 U/L (1.67 U/mg protein). The molecular masses of the enzymes were predicted as about 85 kDa and 110 kDa by SDS-PAGE and zymogram. The enzymatic activity was optimal at $30^{\circ}C$ and the relative agarase activity was decreased as temperature increase from $30^{\circ}C$ and thus about 90% and 70% activities were shown at $40^{\circ}C$ and $50^{\circ}C$, respectively. The optimum pH was 6.0 for agarase activity in 20 mM Tris-HCl buffer and activities were less than 70% and 85% activity at pH 5.0 and pH 7.0, respectively, compared with that at pH 6. Agarase activity has remained over 90% at $20^{\circ}C$ after 1.5 hour exposure at this temperature. However, its activity was less than 60% at $30^{\circ}C$ after 0.5 h exposure at this temperature. The enzymes produced agarooligosaccharides such as agaropentaose and agarotriose from agarose, indicating that the agarases are ${\alpha}$-agarases. Thus, Alteromonas sp. SH-1 and its agarases would be useful for the industrial production of agarooligosaccharides which are known as having anticancer and antioxidation activities.
Keywords
${\alpha}$-Agarase; agaropentaose; agarooligosaccharide; Alteromonas sp. SH-1; marine derived bacterium;
Citations & Related Records
Times Cited By KSCI : 5  (Citation Analysis)
연도 인용수 순위
1 Jang, H. J., D. G. Lee, S. W. Lee, M. J. Jeon, W. J. Chun, K. K. Kwon, H. S. Lee, and S. H. Lee (2011) Isolation of a marinederived Flammeovirga sp. mbrc-1 strain and characterization of its agarase. KSBB J. 26: 552-556.   DOI
2 Lee, D. G., N. Y. Kim, M. K. Jang, O. H. Lee, 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.   DOI
3 Hong, J. H., J. J. Lee, S. H. Hur, H. S. Choi, and J. Y. Kong (2001) Effect of agarooligosaccharides on the growth of intestinal bacteria. J. Fd. Hyg. Safety 16: 11-15.
4 Seo, Y. B., Y. Lu, W. J. Chi, H. R. Pack, K. J. Jeong, S. K. Hong, and Y. K. Chang (2014) Heterologous expression of a newly screened ${\beta}$-agarase from Alteromonas sp. GNUM1 in Escherichia coli and its application for agarose degradation. Process Biochem. 49: 430-436.   DOI
5 Yang, M., X. Mao, N. Liu, Y. Qiu, and C. Xue (2014) Purification and characterization of two agarases from Agarivorans albus OAY 02. Process Biochem. 49: 905-912.   DOI
6 Hassairi, I., R. Ben Amar, M. Nonus, and B. B. Gupta (2001) Production and separation of ${\alpha}$-agarase form Alteromonas agarlyticus strain GJ1B. Biores. Technol. 79: 47-51.   DOI
7 Kim, Y. J. (2010) Properties of the agarase and its gene isolated from a marine bacterium, Tamlana agarivorans. M.S. dissertation, University of Chungnam, Taejon, Korea.
8 Kim, Y. N. (2011) Isolation and purification of new agarase from marine bacterium. M.S. dissertation, University of Kyungsang, Jinju, Korea.
9 Lee, D. G., M. K. Jang, O. H. Lee, N. Y. Kim, S. A. Ju, and S. H. Lee (2008) Over-production of a glycoside hydrolase family 50 ${\beta}$-agarase from Agarivorans sp. JA-1 in Bacillus subtilis and the whitening effect of its product. Biotechnol. Lett. 30: 911-918.   DOI
10 Kim, H. T., S. Lee, D. Lee, H. S. Kim, W. G. Bang, K. H. Kim, and I. G. Choi (2010) Overexpression and molecular characterization of Aga50D from Saccharophagus degradans 2-40: An exo-type ${\beta}$-agarase producing neoagarobiose. Appl. Microbiol. Biotechnol. 86: 227-234.   DOI
11 Lakshmikanth, M., S. Manohar, Y. Souche, and J. Lalitha (2009) Extracellular ${\beta}$-agarase LSL-1 producing neoagarobiose from a newly isolated agar-liquefying soil bacterium, Acinetobacter sp., AG LSL-1. Process Biochem. 44: 999-1003.   DOI
12 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.   DOI
13 Temuujin, U., W. J. Chi, Y. K. Chang, and S. K. Hong (2012) Identification and biochemical characterization of Sco3487 from Streptomyces coelicolor A3(2), an exo- and endo-type ${\beta}$-agarase-producing neoagarobiose. J. Bacteriol. 194: 142-149.   DOI
14 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.   DOI
15 Jang, M. K., S. W. Lee, D. G. Lee, N. Y. Kim, K. H. Yu, H. J. Jang, S. Kim, A. Kim, and S. H. Lee (2010) Enhancement of the thermostability of a recombinant ${\beta}$-agarase, AgaB, from Zobellia galactanivorans by random mutagenesis. Biotechnol. Lett. 32: 943-949.   DOI
16 Wang, J., H. Mou, X. Jiang, and H. Guan (2006) Characterization of a novel ${\beta}$-agarase from marine Alteromonas sp. SY37-12 and its degrading products. Appl. Microbiol. Biotechnol. 71: 833-839.   DOI
17 Chi, W. J., J. H. Lim, D. Y. Park, M. C. Kim, C. J. Kim, Y. K. Chang, and S. K. Hong (2013) Isolation and characterization of a novel agar degrading bacterium Alteromonas macleodii subsp. Gnum08120, from red macroalgae. Korean J. Microbiol. Biotechnol. 41: 8-16.   DOI
18 Kim. J. H., and S. K. Hong (2012) Isolation and characterization of an agarase-producing bacterial strain, Alteromonas sp. GNUM-1, from the west Sea, Korea. J. Microbiol. Biotech. 22: 1621-1628.   DOI
19 Leon, O., L. Quintana, G. Peruzzo and J. C. Slebe (1992) Purification and properties of an extracellular agarase from Alteromonas sp. strain C-1. Microbiol. 58: 4060-4063.
20 Ohta, Y., Y. Hatada, M. Miyazaki, Y. Nogi, S. Ito, and K. Horikoshi (2005) Purification and characterization of a novel from a Thalassomonas sp. Curr. Microbiol. 50: 212-216.   DOI
21 Fu, X. T. and S. M. Kim (2010) Agarase: Review of major sources, categories, purification method, enzyme characteristics and applications. Mar. Drugs 8: 200-218.   DOI
22 Kim, J. H., Y. H. Kim., S. K. Kim, B. W. Kim, and S. W. Nam (2011) Properties and Industrial applications of seaweed polysaccharides-degrading enzymes from the marine microorganisms. Korean J. Microbiol. Biotechnol. 39: 189-199.
23 Kohtaro, K., M. 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-l. J. Biosci. Bioengi. 87: 436-441.   DOI