新型壳聚糖酶CoA制备活性壳寡糖及其在真菌病害防治中的应用
Preparation of Active Chitooligosaccharides with a Novel Chitosanase CoA and Their Application in Fungal Disease Protection.
作者信息
Wang Yanxin, Li Ding, Liu Muxing, Xia Chengyao, Fan Qiwen, Li Xu, Lan Zejun, Shi Guolong, Dong Weiliang, Li Zhoukun, Cui Zhongli
机构信息
Key Laboratory of Agricultural Environmental Microbiology, Ministry of Agriculture and Rural Affairs, College of Life Sciences of Nanjing Agricultural University, 210095 Nanjing, P. R. China.
Institute of Veterinary Immunology & Engineering, Jiangsu Academy of Agricultural Sciences, 210014 Nanjing, P. R. China.
出版信息
J Agric Food Chem. 2021 Mar 24;69(11):3351-3361. doi: 10.1021/acs.jafc.0c07802. Epub 2021 Mar 10.
Enzymes that degrade fungal cell walls and the resulting oligosaccharides are promising weapons to combat plant fungal disease. In this study, we identified a novel endo-chitosanase, CoA, from sp. A7-Y. The enzyme showed a specific activity of 18 U/mg toward 95% deacetylated chitosan at pH 5.0 and 40 °C. CoA also showed activity toward sodium carboxymethylcellulose, indicating substrate promiscuity. CoA hydrolyzed chitosan into chitooligosaccharides (CoA-COSs) with degrees of polymerization (DPs) of 3-5 but showed no activity toward CoA-COSs with DPs <6, indicating an endo-type activity. At 2.5 μg/mL, CoA inhibited appressorium formation of ; the produced CoA-COSs also inhibited the growth of and . Furthermore, CoA-COSs acted as immune elicitors in rice by inducing the reactive oxygen species burst and the expression of defense genes. These results demonstrated that CoA and its resulting CoA-COSs might be effective tools for protecting plants against pathogenic fungi.
降解真菌细胞壁的酶及由此产生的寡糖是对抗植物真菌病害的有力武器。在本研究中,我们从A7-Y菌株中鉴定出一种新型内切壳聚糖酶CoA。该酶在pH 5.0和40℃条件下对95%脱乙酰化壳聚糖的比活性为18 U/mg。CoA对羧甲基纤维素钠也有活性,表明其具有底物混杂性。CoA将壳聚糖水解为聚合度(DPs)为3-5的壳寡糖(CoA-COSs),但对DPs<6的CoA-COSs无活性,表明其具有内切型活性。在2.5μg/mL时,CoA抑制稻瘟病菌附着胞的形成;产生的CoA-COSs也抑制稻瘟病菌和立枯丝核菌的生长。此外,CoA-COSs通过诱导活性氧爆发和防御基因表达,在水稻中作为免疫激发子发挥作用。这些结果表明,CoA及其产生的CoA-COSs可能是保护植物免受病原真菌侵害的有效工具。