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利用新型壳聚糖酶AqCsn1制备低聚合度且具有抗菌特性的壳寡糖。

Preparation of chitooligosaccharides with a low degree of polymerization and anti-microbial properties using the novel chitosanase AqCsn1.

作者信息

Xia Chengyao, Li Ding, Qi Mengyi, Wang Yanxin, Zhang Yue, Yang Yiheng, Hu Zejia, Du Xin, Zhao Yuqiang, Yu Kuai, Huang Yan, Li Zhoukun, Ye Xianfeng, Cui Zhongli

机构信息

Key Laboratory of Agricultural Environmental Microbiology, Ministry of Agriculture, College of Life Science, Nanjing Agricultural University, Nanjing, 210095, PR China.

Institute of Veterinary Immunology & Engineering, Jiangsu Academy of Agricultural Sciences, 210014, Nanjing, PR China; Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, 225009, China; Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base, Ministry of Science and Technology, Nanjing, 210014, China.

出版信息

Protein Expr Purif. 2023 Mar;203:106199. doi: 10.1016/j.pep.2022.106199. Epub 2022 Nov 11.

Abstract

Chitosanases hydrolyze chitosan into chitooligosaccharides (COSs) with various biological activities, which are widely employed in many areas including plant disease management. In this study, the novel chitosanase AqCsn1 belonging to the glycoside hydrolase family 46 (GH46) was cloned from Aquabacterium sp. A7-Y and heterologously expressed in Escherichia coli BL21 (DE3). AqCsn1 displayed the highest hydrolytic activity towards chitosan with 95% degree of deacetylation at 40 °C and pH 5.0, with a specific activity of 13.18 U/mg. Product analysis showed that AqCsn1 hydrolyzed chitosan into (GlcN) and (GlcN) as the main products, demonstrating an endo-type cleavage pattern. Evaluation of antagonistic activity showed that the hydrolysis products of AqCsn1 suppress the mycelial growth of Magnaporthe oryzae and Phytophthora sojae in a concentration-dependent manner, and the inhibition rate of P. sojae reached 39.82% at a concentration of 8 g/L. Our study demonstrates that AqCsn1 and hydrolysis products with a low degree of polymerization might have potential applications in the biological control of agricultural diseases.

摘要

壳聚糖酶将壳聚糖水解为具有多种生物活性的壳寡糖(COSs),壳寡糖在包括植物病害管理在内的许多领域都有广泛应用。在本研究中,从水生杆菌属菌株A7-Y中克隆了属于糖苷水解酶家族46(GH46)的新型壳聚糖酶AqCsn1,并在大肠杆菌BL21(DE3)中进行了异源表达。AqCsn1在40℃和pH 5.0条件下对脱乙酰度为95%的壳聚糖表现出最高的水解活性,比活性为13.18 U/mg。产物分析表明,AqCsn1将壳聚糖水解为以(GlcN)和(GlcN)为主的产物,呈现内切型切割模式。拮抗活性评估表明,AqCsn1的水解产物以浓度依赖的方式抑制稻瘟病菌和大豆疫霉的菌丝生长,在浓度为8 g/L时对大豆疫霉的抑制率达到39.82%。我们的研究表明,AqCsn1和低聚合度的水解产物在农业病害生物防治中可能具有潜在应用。

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