Biochemical Characterization of a Novel Acidic Exochitinase from Rhizomucor miehei with Antifungal Activity

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• 作者：Shaoqing Yang ; Xin Fu ; Qiaojuan Yan ; Zhengqiang Jiang ; Jing Wang
• 刊名：Journal of Agricultural and Food Chemistry
• 出版年：2016
• 出版时间：January 20, 2016
• 年：2016
• 卷：64
• 期：2
• 页码：461-469
• 全文大小：732K
• ISSN：1520-5118

文摘

A novel chitinase gene (RmChi44) from Rhizomucor miehei was cloned and expressed in Escherichia coli as an intracellular soluble and active protein. The recombinant chitinase (RmChi44) was purified to homogeneity and biochemically characterized. The molecular mass of RmChi44 was estimated to be 44.6 kDa on SDS-PAGE. RmChi44 displayed an acidic pH optimum of 4.5 and was stable within pH 4.5–9.0. The optimal temperature of RmChi44 was found to be 50 °C. The K_m values of RmChi44 for colloidal chitin and glycol chitin were 4.02 and 1.55 mg/mL, respectively. RmChi44 hydrolyzed colloidal chitin to yield mainly N-acetyl chitobiose, exhibiting an exotype cleavage pattern. Moreover, the enzyme displayed β-N-acetylglucosaminidase activity, splitting N-acetyl COSs with degree of polymerization (DP) 2–5 into their monomer. In addition, RmChi44 showed antifungal activity against some phytopathogenic fungi. This is the first report on an exochitinase showing β-N-acetylglucosaminidase activity and antifungal activity from Rhizomucor species.