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KMID : 0525619970020030040
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1997 Volume.2 No. 3 p.40 ~ p.48
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Development of Chitosanase for the Production of Chitooligosaccharides
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최연진/Choi YJ
김은정/김영수/신용철/Kim EJ/Kim YS/Shin YC
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Abstract
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For the enzymatic production of chitooligosaccharides from chitosan, an endo-chitosanase producing bacterium, Bacillus sp. GM44 was isolated from a soil. The bacterium produced 50U/ml of extracellular chitosanase constitutively. Chitosanase was purified from culture supernatant of strain GM44 by CM-Toyopearl column chromatography and Superose 12HR column chromatography. Molecular mass of the enzyme was estimated as 45,000 by SDS-PAGE. Its optimum pH and temperature were 5.0 and 70t, respectively. It was stable in the pH range of 3.0 to 10.0 and up to 40℃. Its N-terminal amino acid sequence had 90% similarity with an endo-cellulase of Bacillus sp. KSM-330. The enzyme showed high substrate specificity toward chitosan but could hydrolyze swollen chitin and CM-cellulose with a low hydrolysis rate. Chitobiose, chitotriose, chitotetraose, and chitopentaose were fairly resistant to the enzyme action and while, chitohexaose and the higher chitooligosaccharides were hydrolyzed easily. The enzyme had maximum activity against 81.9% deacetylated chitosan and also showed 83.3% activity against even 39.2% deacetylated chitosan. It produced chitooligosaccharides ranging from chitotriose to chitooctaose as major end-products from chitosan.
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