Characterization of an alkalophilic extracellular chitosanase from Bacillus cereus GU-02

被引:16
|
作者
Goo, Bon Geun [1 ]
Park, Jae Kweon [2 ]
机构
[1] Gachon Univ, Dept Pharmaceut Sci, Inchon 406799, South Korea
[2] Gachon Univ, Dept Life Sci, Coll Bionano, Inchon 406799, South Korea
关键词
Bacillus cereus; Alkalophilic chitosanase; Chitosan; Chitooligosaccharides; Kinetic properties; HEXA-N-ACETYLCHITOHEXAOSE; PURIFICATION; CHITIN; OLIGOSACCHARIDES; SPECIFICITY; MODE;
D O I
10.1016/j.jbiosc.2013.11.005
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
An alkalophilic extracellular chitosanase (ACTase) was characterized from the culture supernatant of Bacillus cereus GU-02. Kinetic properties of ACTase produced from B. cereus GU-02 after cultivation in anaerobic condition, alkaline medium (pH 10) at 37 degrees C for 3 days were investigated. ACTase was found to be stable in alkaline pH range from 8 to 10. Interestingly, optimum pH and temperature were estimated to be 10 and 37 degrees C, respectively, where ACTase showed chitosan degrading activity (87%), which was enhanced by 15% in the presence of calcium ions (8 mM). The ACTase produced from B. cereus GU-02 was partially purified from the culture supernatant, and its enzymatic activity was kinetically characterized. The V-max and K-m were estimated with a chitosan (degree of deacetylation, DD 92% as substrate) as 0.038 U/min/mu g protein and 0327 mu M, respectively. A combination of the TIC and MALDI-TOF MS results showed that the chitosan oligosaccharides obtained from the hydrolysis of high molecular weight chitosan (HMWC) by ACTase of the B. cereus GU-2 comprise oligomers with degree of polymerization (DP) mainly from dimers to pentamers. High production of ACTase and chitooligosaccharides may be useful for various industrial and biological applications. (C) 2013, The Society for Biotechnology, Japan. All rights reserved.
引用
收藏
页码:684 / 689
页数:6
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