Identification, expression, and characterization of a marine-derived chitinase Ce 0303 from Chitiniphilus eburneus YS-30 with exo- and endo- hydrolytic properties

文献类型： 外文期刊

第一作者： Liu, Xiaofang

作者： Liu, Xiaofang;Miao, Junkui;Wang, Xixi;Leng, Kailiang;Chen, Sunan;Yan, Mingyan;Li, Yinping;Yang, Min;Xiao, Mengshi;Mou, Haijin;Leng, Kailiang;Mou, Haijin;Leng, Kailiang

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关键词： Chitinase; N -acetyl-oligosaccharides; Crystalline chitin

期刊名称：INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES （ 影响因子：7.7； 五年影响因子：7.7 ）

ISSN： 0141-8130

年卷期： 2024 年 276 卷

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收录情况： SCI

摘要： N-acetyl-oligosaccharides exhibit antioxidant and antibacterial activities. However, the low catalytic efficiency of chitinase on crystalline chitin hinders the eco-friendly production of N-acetyl-oligosaccharides. A marinederived chitinase-producing strain Chitiniphilus eburneus YS-30 was screened in this study. The genome of C. eburneus YS-30 spans 4,522,240 bp, with a G + C content of 63.96 % and 4244 coding genes. Among the chitinases secreted by C. eburneus YS-30, Ce0303 showed the highest content at 19.10 %, with a molecular weight of 73.5 kDa. Recombinant Ce0303 exhibited optimal activity at 50 degrees C and pH 5.0, maintaining stability across pH 4.0-10.0. Ce0303 demonstrated strict substrate specificity, with a specific activity toward colloidal chitin of 6.41 U mg-1, Km of 2.34 mg mL-1, and kcat of 3.27 s-1. The specific activity of Ce0303 toward alpha-chitin was 18.87 % of its activity on colloidal chitin. Ce0303 displayed both exo- and endo-hydrolytic properties, primarily producing (GlcNAc)1-3 from colloidal chitin. The structure of Ce0303 includes one catalytic domain and two chitinbinding domains. Docking results revealed that the GlcNAc at -1 subsite formed two hydrogen bonds with conserved Trp380. The hydrolytic properties of Ce0303 will provide technical support for the comprehensive utilization of crustacean raw materials.

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