文献类型：期刊文献

英文题名：Site-Specific and Covalent Immobilization of Lipase on Natural Polyphenol-Modified Magnetic Nanoparticles for Effective Biodiesel Production

作者：Tang, Wen[1,2];Li, Haoxiang[1];Zhang, Wei[1];Ma, Tonghao[1];Zhuang, Jiafeng[1];Wang, Ping[3];Chen, Chao[1]

机构：[1]East China Univ Sci & Technol, Shanghai Collaborat Innovat Ctr Biomfg Technol, Sch Biotechnol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Shanghai Inst Pharmaceut Ind, China State Inst Pharmaceut Ind, Shanghai 201203, Peoples R China;[3]Univ Minnesota, Dept Bioprod & Biosyst Engn, St Paul, MN 55108 USA

年份：2022

卷号：10

期号：17

起止页码：5384

外文期刊名：ACS SUSTAINABLE CHEMISTRY & ENGINEERING

收录：;EI(收录号：20221712033572);WOS:【SCI-EXPANDED(收录号:WOS:000841791200002)】；

基金：This work was sponsored by the National Natural Science Foundation of China (nos. 21908059 and 21636003), the China Postdoctoral Science Foundation (no. 2019M651419), Shanghai Sailing Program (no. 19YF1410900), the Natural Science Foundation of Shanghai (22ZR1415400), the Fundamental Research Funds for the Central Universities (no. 22221818014), the Shanghai Postdoctoral Excellence Program (no. 2018011), the Foundation of State Key Laboratory of Biobased Material and Green Papermaking, Qilu University of Technology, Shandong Academy of Sciences (grant no.: GZKF202031), and the Open Funding Project of the State Key Laboratory of Bioreactor Engineering.

语种：英文

外文关键词：site-specific immobilization; activity recovery; 3D structural analysis; biodiesel production; green manufacturing

摘要：Site-specific and covalent attachment is the most desirable immobilization strategy, but classic methods typically require genetic engineering or complicated material fabrication, resulting in operational complexity and difficulty. Herein, a novel site-specific and covalent immobilization strategy based on accurately selected immobilization sites on lipase was developed. Specifically, computer-aided structural analysis of functional groups on lipase revealed that lysine residues with free amino groups were far away from the catalytic pocket and lid, which were suitable to be chosen as the best immobilization sites to effectively reduce the loss in its activity. Meanwhile, natural polyphenol-modified magnetic nanoparticles could increase the active immobilization sites, and lipase can be immobilized on them directly via a covalent reaction. This site-specific immobilization system exhibited significant enhancement in activity recovery (71.3%) compared to random immobilization (42.5 and 55.9%). As expected, experimental and computational analyses revealed that tailor-made site-specific immobilization carriers were beneficial to maintain the native catalytic pocket conformation and enhance the rigidity of the immobilized lipase, which exhibited a higher biodiesel yield (92.1%) than free and randomly immobilized lipases. Besides, the site-specifically immobilized lipase could maintain as high as 75.3% biodiesel yield after eight cycles, making it an ideal nanocatalyst for efficient production of biodiesel. Overall, the site-specific enzyme immobilization technology can provide stable catalytic activity advantages over the randomly covalent immobilization strategy, which can significantly promote green manufacturing and sustainable production.