文献类型：期刊文献

英文题名：Virtual screening of carboxylic acid reductases for biocatalytic synthesis of 6-aminocaproic acid and 1,6-hexamethylenediamine

作者：Shi, Kun[1];Li, Ju-Mou[1];Zhang, Zhi-Jun[1];Chen, Qi[1];Xu, Jian-He[1];Yu, Hui-Lei[1,2]

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

年份：2023

卷号：120

期号：7

起止页码：1773

外文期刊名：BIOTECHNOLOGY AND BIOENGINEERING

收录：;EI(收录号：20231914066499);WOS:【SCI-EXPANDED(收录号:WOS:000980233400001)】；

基金：This work was financially supported by the National Key Research and Development Program of China (2021YFC2102900 & 2019YFA0905000), the National Natural Science Foundation of China (grant numbers 21922804 & 32271540), Program of Shanghai Academic Research Leader (21XD1400800), and Frontiers Science Center for Materiobiology and Dynamic Chemistry JKVF12001035.

语种：英文

外文关键词：1; 6-hexamethylenediamine; 6-aminocaproic acid; adipic acid; carboxylic acid reductase; virtual screening

摘要：The key precursors for nylon synthesis, that is, 6-aminocaproic acid (6-ACA) and 1,6-hexamethylenediamine (HMD), are produced from petroleum-based feedstocks. A sustainable biocatalytic alternative method from bio-based adipic acid has been demonstrated recently. However, the low efficiency and specificity of carboxylic acid reductases (CARs) used in the process hampers its further application. Herein, we describe a highly accurate protein structure prediction-based virtual screening method for the discovery of new CARs, which relies on near attack conformation frequency and the Rosetta Energy Score. Through virtual screening and functional detection, five new CARs were selected, each with a broad substrate scope and the highest activities toward various di- and omega-aminated carboxylic acids. Compared with the reported CARs, KiCAR was highly specific with regard to adipic acid without detectable activity to 6-ACA, indicating a potential for 6-ACA biosynthesis. In addition, MabCAR3 had a lower K-m with regard to 6-ACA than the previously validated CAR MAB4714, resulting in twice conversion in the enzymatic cascade synthesis of HMD. The present work highlights the use of structure-based virtual screening for the rapid discovery of pertinent new biocatalysts.