文献类型：期刊文献

英文题名：Binding Peptide-Guided Immobilization of Lipases with Significantly Improved Catalytic Performance Using Escherichia coli BL21(DE3) Biofilms as a Platform

作者：Dong, Hao[1];Zhang, Wenxue[1];Xuan, Qize[1];Zhou, Yao[1];Zhou, Shengmin[1];Huang, Jiaofang[1];Wang, Ping[2]

机构：[1]East China Univ Sci & Technol, Sch Biotechnol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Univ Minnesota, Dept Bioprod & Biosyst Engn, St Paul, MN 55108 USA

年份：2021

卷号：13

期号：5

起止页码：6168

外文期刊名：ACS APPLIED MATERIALS & INTERFACES

收录：;EI(收录号：20210709915428);WOS:【SCI-EXPANDED(收录号:WOS:000619638400025)】；

基金：This work was sponsored by the National Natural Science Foundation of China (21636003, 21672065, 22077032, and 31872728), the International S&T Innovation Cooperation Key Project (2017YFE0129600), the National Science and Technology Major Project of the Ministry of Science and Technology of China (2020YFA0908900), the Natural Science Foundation of Shanghai (19ZR1477100), the Fundamental Research Funds for the Central Universities (22221818014), and the 111 Project (B18022).

语种：英文

外文关键词：biofilm; lipase; lipase binding peptide; immobilization; phthalic acid esters

摘要：Developing novel immobilization methods to maximize the catalytic performance of enzymes has been a permanent pursuit of scientific researchers. Engineered Escherichia coli biofilms have attracted great concern as surface display platforms for enzyme immobilization. However, current biological conjugation methods, such as the SpyTag/SpyCatcher tagging pair, that immobilize enzymes onto E. coli biofilms seriously hamper enzymatic performance. Through phage display screening of lipase-binding peptides (LBPs) and co-expression of CsgB (nucleation protein of curli nanofibers) and LBP2-modified CsgA (CsgALBP2, major structural subunit of curli nanofibers) proteins, we developed E. coli BL21::Delta CsgA-CsgB-CsgALBP2 (LBP2-functionalized) biofilms as surface display platforms to maximize the catalytic performance of lipase (Lip181). After immobilization onto LBP2-functionalized biofilm materials, Lip181 showed increased thermostability, pH, and storage stability. Surprisingly, the relative activity of immobilized Lip181 increased from 8.43 to 11.33 U/mg through this immobilization strategy. Furthermore, the highest loading of lipase on LBP2-functionalized biofilm materials reached up to 27.90 mg/g of wet biofilm materials, equivalent to 210.49 mg/g of dry biofilm materials, revealing their potential as a surface with high enzyme loading capacity. Additionally, immobilized Lip181 was used to hydrolyze phthalic acid esters, and the hydrolysis rate against dibutyl phthalate was up to 100%. Thus, LBP2-mediated immobilization of lipases was demonstrated to be far more advantageous than the traditional SpyTag/SpyCatcher strategy in maximizing enzymatic performance, thereby providing a better alternative for enzyme immobilization onto E. coli biofilms.