文献类型：期刊文献

英文题名：Computation-driven redesign of an NRPS-like carboxylic acid reductase improves activity and selectivity

作者：Shi, Kun[1];Li, Ju-Mou[1];Wang, Mu-Qiang[1];Zhang, Yi-Ke[1];Zhang, Zhi-Jun[1];Chen, Qi[1];Hollmann, Frank[2];Xu, Jian-He[1];Yu, Hui-Lei[1]

机构：[1]East China Univ Sci & Technol, Sch Biotechnol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]Delft Univ Technol, Dept Biotechnol Inst, Maasweg 9, NL-2629HZ Delft, Netherlands

年份：2024

卷号：10

期号：48

外文期刊名：SCIENCE ADVANCES

收录：;EI(收录号：20245017503211);WOS:【SCI-EXPANDED(收录号:WOS:001402032000002)】；

基金：This work was supported by the National Key Research and Development Program of China 2019YFA0905000 (to H.-L.Y.) and 2021YFC2102900 (to H.- L.Y.), the Shanghai Commission of Science and Technology 23HC1400200 (to H.- L.Y.),the National Natural Science Foundation of China 32401276 (to K.S.) , 21922804 (to H.- L.Y.), and 32271540 (to H.-L.Y.), the Program of Shanghai Academic Research Leader 21XD1400800 (to H.- L.Y.), and the Frontiers Science Center for Materiobiology and Dynamic Chemistry JKVF12001035 (to J.- H.X.).

语种：英文

外文关键词：Biosynthesis - Carboxylation

摘要：Engineering nonribosomal peptide synthetases (NRPSs) has been a "holy grail" in synthetic biology due to their modular nature and limited understanding of catalytic mechanisms. Here, we reported a computational redesign of the "gate-keeper" adenylation domain of the model NRPS-like enzyme carboxylic acid reductases (CARs) by using approximate mechanism-based geometric criteria and the Rosetta energy score. Notably, MabCAR3 mutants ACA-1 and ACA-4 displayed a remarkable improvement in catalytic efficiency (kcat/KM) for 6-aminocaproic acid, up to 101-fold. Furthermore, G418K exhibited an 86-fold enhancement in substrate specificity for adipic acid compared to 6-aminocaproic acid. Our work provides not only promising biocatalysts for nylon monomer biosynthesis but also a strategy for efficient NRPSs engineering.