文献类型：期刊文献

英文题名：Heterologous Expression and Functional Characterization of a Truncated Marine Alginate Lyase

作者：Shao, Qianqian[1];Yao, Chaoying[1];Wang, Xingbin[1];Han, Bei[1];Liu, Qi[1];Cai, Menghao[1,2]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai, Peoples R China;[2]East China Univ Sci & Technol, Shanghai Collaborat Innovat Ctr Biomfg, Shanghai 200237, Peoples R China

年份：2025

卷号：197

期号：11

起止页码：7694

外文期刊名：APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY

收录：;EI(收录号：20254219338826);WOS:【SCI-EXPANDED(收录号:WOS:001592711800001)】；

基金：This study was supported by the National Key R&D Program of China (2022YFC2805102), National Natural Science Foundation of China (32201206), China Postdoctoral Science Foundation (2022M711146), and the 111 project (B18022).

语种：英文

外文关键词：Alginate lyase; Komagataella phaffii; Non-catalytic domain truncation; Enzymatic property; Heterologous expression

摘要：Alginate lyase is widely used in the preparation of alginate oligosaccharides, medicine production, energy conversion, and so on. In this study, we designed truncated mutants of a marine-derived alginate lyase Algl and identified a highly active mutant CD317 with less degradation when expressed in a yeast host. The enzyme activity of the secretory CD317 was 1.4-fold higher than that of the parent Algl. It degraded both polyM and polyG but had a stronger preference for polyM. The optimal temperature of Algl and CD317 was 40 degrees C and 35 degrees C respectively, for which CD317 showed better temperature tolerance. Additionally, Ca2+ highly improved the enzyme activity. Fermentation conditions for CD317 production were optimized as a culture time of 144 h, an inoculum of an OD600nm of 0.5, and an inducer concentration of 2% (v/v), respectively. Bioreactor fermentation allowed the highest production of 19,500 U/mL, which was 4.6-fold higher than that in a well-plate. These results indicated that the truncated CD317 showed good potential for industrial use.