文献类型：期刊文献

英文题名：Bioconversion of chitin into chitin oligosaccharides using a novel chitinase with high chitin-binding capacity

作者：Zhao, Qiong[1,2];Fan, Liqiang[1,2];Deng, Chen[1,2];Ma, Chunyu[1,2];Zhang, Chunyue[1,4];Zhao, Liming[1,2,3,4]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Shanghai Collaborat Innovat Ctr Biomfg Technol SCI, Shanghai 200237, Peoples R China;[3]East China Univ Sci & Technol, Shanghai Frontiers Sci Ctr Optogenet Tech Cell Met, Sch Pharm, 130 Mei Long Rd, Shanghai 200237, Peoples R China;[4]East China Univ Sci & Technol, Sch Biotechnol, Meilong Rd 130, Shanghai 200237, Peoples R China

年份：2023

卷号：244

外文期刊名：INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES

收录：;EI(收录号：20232414250117);WOS:【SCI-EXPANDED(收录号:WOS:001027214800001)】；

基金：This work was supported by the National Key Research and Development Program of China (2019YFD0901805), the 111 Project (B18022), the Fundamental Research Funds for the Central Universities, the Open Project Funding of the State Key Laboratory of Bioreactor Engineering, ECUST (ZDXM2019), and Shanghai Frontiers Science Center of Optogenetic Techniques for Cell Metabolism (Shanghai Municipal Education Commission).

语种：英文

外文关键词：Chitinase; Chitin oligosaccharides; Purification; Molecular simulation; Biodegradation

摘要：Chitin is the second largest renewable biomass resource in nature, it can be enzymatically degraded into highvalue chitin oligosaccharides (CHOSs) by chitinases. In this study, a chitinase (ChiC8-1) was purified and biochemically characterized, its structure was analyzed by molecular modeling. ChiC8-1 had a molecular mass of approximately 96 kDa, exhibited its optimal activity at pH 6.0 and 50 degrees C. The Km and Vmax values of ChiC8-1 towards colloidal chitin were 10.17 mgmL-1 and 13.32 U/mg, respectively. Notably, ChiC8-1 showed high chitin-binding capacity, which may be related to the two chitin binding domains in the N-terminal. Based on the unique properties of ChiC8-1, a modified affinity chromatography method, which combines protein purification with chitin hydrolysis process, was developed to purify ChiC8-1 while hydrolyzing chitin. In this way, 9.36 & PLUSMN; 0.18 g CHOSs powder was directly obtained by hydrolyzing 10 g colloidal chitin with crude enzyme solution. The CHOSs were composed of 14.77-2.83 % GlcNAc and 85.23-97.17 % (GlcNAc)2 at different enzyme-substrate ratio. This process simplifies the tedious purification and separation steps, and may enable its potential application in the field of green production of chitin oligosaccharides.