Purification of chitinase/chitosanase from Bacillus cereus and discovery of an enzyme inhibitor

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• 作者：Tzu-Wen Liang ; Yue-Yin Chen ; Po-Shen Pan ; San-Lang Wang
• 关键词：Chitinase ; Chitosanase ; Squid pen ; Bacillus cereus ; Chitinase inhibitor ; Boron
• 刊名：International Journal of Biological Macromolecules
• 出版年：February, 2014
• 年：2014
• 卷：63
• 期：Complete
• 页码：8-14
• 全文大小：1081 K

文摘

A chitinase and a chitosanase were induced from a squid pen powder (SPP)-containing medium of Bacillus cereus TKU030 and purified by precipitation with ammonium sulphate and combined column chromatography. The purified chitinase and chitosanase exhibited optimum activity at 60 掳C, pH 5-6 and 40 掳C, pH 4, respectively. The chitinase and chitosanase were stable at 25-60 掳C, pH 4-7 and 25-50 掳C, pH 3-7, respectively. The chitinase and chitosanase showed the highest activity toward 尾-chitin and 60% DD chitosan, respectively. The chitinase was significantly inhibited by Mn²⁺ and EDTA but activated by Cu²⁺, Fe²⁺ and Ca²⁺. The chitosanase was significantly inhibited by Cu²⁺, Fe²⁺, Zn²⁺, Mn²⁺ and EDTA. The chitinase showed high stability in the presence of various surfactants, such as SDS, Tween 20, Tween 40 and Triton X-100. In contrast, these surfactants were inhibitors of the chitosanase. The chitinase and chitosanase were also inhibited by TKUPSP017, a small synthetic boron-containing molecule with a BF₃K side-chain. However, TKUPSP017 enhanced the growth of B. cereus TKU030 in SPP-containing medium.