Design and Synthesis of a Chitodisaccharide-Based Affinity Resin for Chitosanases Purification
文献类型： 外文期刊
作者： Li, Shangyong 1 ; Wang, Linna 2 ; Chen, Xuehong 1 ; Sun, Mi 2 ; Han, Yantao 1 ;
作者机构： 1.Qingdao Univ, Dept Pharmacol, Coll Basic Med, Qingdao 266071, Peoples R China
2.Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, Minist Agr, Key Lab Sustainable Dev Marine Fisheries, Qingdao 266071, Peoples R China
关键词： chitosanases; adsorption analysis; affinity purification
期刊名称：MARINE DRUGS （ 影响因子：5.118； 五年影响因子：5.951 ）
ISSN： 1660-3397
年卷期： 2019 年 17 卷 1 期
页码：
收录情况： SCI
摘要： Chitooligosaccharides (CHOS) have gained increasing attention because of their important biological activities. Enhancing the efficiency of CHOS production essentially requires screening of novel chitosanase with unique characteristics. Therefore, a rapid and efficient one-step affinity purification procedure plays important roles in screening native chitosanases. In this study, we report the design and synthesis of affinity resin for efficient purification of native chitosanases without any tags, using chitodisaccharides (CHDS) as an affinity ligand, to couple with Sepharose 6B via a spacer, cyanuric chloride. Based on the CHDS-modified affinity resin, a one-step affinity purification method was developed and optimized, and then applied to purify three typical glycoside hydrolase (GH) families: 46, 75, and 80 chitosanase. The three purified chitosanases were homogeneous with purities of greater than 95% and bioactivity recovery of more than 40%. Moreover, we also developed a rapid and efficient affinity purification procedure, in which tag-free chitosanase could be directly purified from supernatant of bacterial culture. The purified chitosanases samples using such a procedure had apparent homogeneity, with more than 90% purity and 10-50% yield. The novel purification methods established in this work can be applied to purify native chitosanases in various scales, such as laboratory and industrial scales.
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