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KMID : 0545120110210101021
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Journal of Microbiology and Biotechnology
2011 Volume.21 No. 10 p.1021 ~ p.1025
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Isolation, Purification, and Enzymatic Characterization of Extracellular Chitosanase from Marine Bacterium Bacillus subtilis CH2
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Oh Chul-Hong
Mahanama De Zoysa
Kang Do-Hyung
Lee Young-Deuk
Whang Il-Son
Chamilani Nikapitiya
Heo Soo-Jin
Yoon Kon-Tak
Abu Affan
Lee Je-Hee
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Abstract
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A Bacillus subtilis strain was isolated from the intestine of Sebastiscus marmoratus (scorpion fish) that was identified as Bacillus subtilis CH2 by morphological, biochemical, and genetic analyses. The chitosanase of Bacillus subtilis CH2 was best induced by fructose and not induced with chitosan, unlike other chitosanases. The strain was incubated in LB broth, and the chitosanase secreted into the medium was concentrated with ammonium sulfate precipitation and purified by gel permeation chromatography. The molecular mass of the purified chitosanase was detected as 29 kDa. The optimum pH and temperature of the purified chitosanase were 5.5 and 60oC, respectively. The purified chitosanase was continuously thermostable at 40oC. The specific acitivity of the purified chitosanase was 161 units/mg. The N-terminal amino acid sequence was analyzed for future study.
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KEYWORD
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chitosan, chitosanase, chito-oligosaccharide, Bacillus subtilis, Sebastiscus marmoratus, purification
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