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http://dx.doi.org/10.3839/jabc.2009.027

Isolation of an Agarolytic Bacteria, Cellvibrio mixtus SC-22 and The Enzymatic Properties  

Cha, Jeong-Ah (Department of Bio-Environmental Chemistry, College of Agriculture and Lifesciences, Chungnam National University)
Kim, Yoo-Jin (Department of Bio-Environmental Chemistry, College of Agriculture and Lifesciences, Chungnam National University)
Seo, Yung-Bum (Department of Forest Products, College of Agriculture and Lifesciences, Chungnam National University)
Yoon, Min-Ho (Department of Bio-Environmental Chemistry, College of Agriculture and Lifesciences, Chungnam National University)
Publication Information
Journal of Applied Biological Chemistry / v.52, no.4, 2009 , pp. 157-162 More about this Journal
Abstract
An agar-liquefying bacteria (SC-22), which produces a diffusible agarase that caused agar softening around the colony was isolated from Daecheong lake in Korea. Chemotaxanomic and phylogenetic analyses based on 16S rRNA gene sequences revealed the strain was classified as Cellvibrio mixtus SC-22. The isolate SC-22 showed maximal extracellular agarase activity with 58.5 U/mL after 48 h cultivation in the presence of 0.2% agar. It was observed that the isolate produced two kinds of extracellular and three kinds of intracellular isoenzymes. The major agarase was purified from the culture filtrate of agarolytic bacteria by ammonium sulfate precipitation, anion exchange and gel filtration column chromatographic methods. The molecular mass of the purified enzyme was estimated to be 25 kDa by SDS-PAGE. The optimum pH and temperature of the purified enzyme were pH 7.0 and $50^{\circ}C$, respectively. The agarase activity was activated by $Fe^{2+}$, $Na^+$ and $Ca^{2+}$ ions while it was inhibited by $Hg^{2+}$, $Mn^{2+}$ and $Cu^{2+}$ at 1 mM concentration. The predominant hydrolysis product of agarose by the enzyme was galactose and disaccharide on TLC, indicating the cleavage of $\beta$-1,4 linkage in a random manner. The enzyme showed high substrate specificity for only agar and agarose among various polysaccharides.
Keywords
agarase; characterization; Cellvibrio mixtus SC-22; isozymes; purification;
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