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Thermostable Chitosanase from Bacillus sp. Strain CK4: Its Purification, Characterization, and Reaction Patterns

Authors
 Ho-Geun YOON  ;  Hee-Yun KIM  ;  Hye-Kyung KIM  ;  Bum-Shik HONG  ;  Dong-Hoon SHIN  ;  Hong-Yon CHO 
Citation
 BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, Vol.65(4) : 802-809, 2001 
Journal Title
 BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 
ISSN
 0916-8451 
Issue Date
2001
MeSH
Ammonium Sulfate/chemistry ; Bacillus/enzymology* ; Chromatography, Gel ; Chromatography, Ion Exchange ; Chromatography, Thin Layer ; Glycoside Hydrolases/chemistry* ; Hydrogen-Ion Concentration ; Isoelectric Focusing ; Metals/chemistry ; Molecular Weight ; Oligosaccharides/chemistry ; Substrate Specificity ; Temperature
Keywords
Thermostable chitosanase ; Bacillus sp.strain CK4 ; Chitosan oligosaccharide
Abstract
A thermostable chitosanase, purified 156-fold to homogeneity in an overall yield of 12.4%, has a molecular weight of about 29,000±2,000, and is composed of monomer. The enzyme degraded soluble chitosan, colloidal chitosan, and glycol chitosan, but did not degrade chitin or other β-linked polymers. The enzyme activity was increased about 2.5-fold by the addition of 10 mM Co2+ and 1.4-fold by Mn2+. However, Cu2+ ion strongly inhibited the enzyme. Optimum temperature and pH were 60°C and 6.5, respectively. The enzyme was stable after heat treatment at 80°C for 30 min or 70°C for 60 min and fairly stable in protein denaturants as well. Chitosan was hydrolyzed to (GlcN)4 as a major product, by incubation with the purified enzyme. The effects of ammonium sulfate and organic solvents on the action pattern of the thermostable chitosanase were investigated. The amounts of (GlcN)3-(GlcN)6 were increased about 30% (w/w) in DAC 99 soluble chitosan containing 10% ammonium sulfate, and (GlcN)1 was not produced. The monophasic reaction system consisted of DAC 72 soluble chitosan in 10% EtOH also showed no formation of (GlcN)1, however, the yield of (GlcN)3 ∼ (GlcN)6 was lower than DAC 99 soluble chitosan-10% ammonium sulfate. The optimal concentration of ammonium sulfate to be added was 20%. At this concentration, the amount of hexamer was increased by over 12% compared to the water-salt free system.
Files in This Item:
T200101185.pdf Download
DOI
10.1271/bbb.65.802
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Biochemistry and Molecular Biology (생화학-분자생물학교실) > 1. Journal Papers
Yonsei Authors
Yoon, Ho Geun(윤호근) ORCID logo https://orcid.org/0000-0003-2718-3372
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/141960
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