文献类型：期刊文献

中文题名：脂肪酶 RWL 催化合成二氢燕麦生物碱 D

英文题名：Catalytic synthesis of dihydroavenanthramide D by lipase RWL

作者：Chen, Jinbo[1]; Zhai, Chuntao[2]; Jin, Minrong[2]; Wu, Qian[3]; Yuan, Minjia[4]; Wang, Xuedong[1]; Ren, Yuhong[1]; Wang, Hualei[1]

机构：[1] School of Biotechnology, East China University of Science and Technology, Shanghai, 200237, China; [2] Shanghai Zhina Biotechnology Co., Ltd., Shanghai, 201400, China; [3] Shanghai Qiran Biotechnology Co., Ltd., Shanghai, 201702, China; [4] Shanghai Jinjia Technology Co., Ltd., Shanghai, 201210, China

年份：2026

卷号：42

期号：6

起止页码：2687

外文期刊名：Shengwu Gongcheng Xuebao/Chinese Journal of Biotechnology

收录：EI(收录号：20263021183942);Scopus(收录号：2-s2.0-105043669168)

语种：中文

外文关键词：Amides - Aniline - Aspartame - Biochemical engineering - Biosynthesis - Catalysis - Esters - Genetic engineering - Methyl ester - Plants (botany)

摘要：Dihydroavenanthramide D (DHAvD) is an aromatic amide with anti-inflammatory, antipruritic, and antioxidant properties. Its production currently relies on chemical synthesis. Lipases can catalyze the aminolysis of esters or acids to synthesize a series of amide compounds. However, the enzymatic synthesis of DHAvD is particularly challenging due to the ortho-substituted carboxyl group on its amine donor, 2-aminobenzoic acid (2-AA). Addressing this challenge, this study aims to identify a novel lipase capable of tolerating ortho-substituted anilines and to establish a suitable DHAvD-catalyzed synthetic system. Through genomic mining, this study successfully identified lipase RWL from Rhizorhabdus wittichii, which utilizes 2-AA as the amine donor and either methyl 3- (4-hydroxyphenyl)propionate (MHPP) or p-hydroxyphenylpropionic acid (PHBPA)as the acyl donor to synthesize DHAvD in aqueous phase. To enhance reaction efficiency, we established the optimal organic-phase reaction systems for both carboxylic acid and ester substrates: hexane for MHPP and methyl tert-butyl ether for PHPA. This switch from aqueous phase increased the DHAvD yield from 0.1% to 3.9%. Building on this foundation, molecular docking-guided alanine scanning and saturation mutagenesis were employed to modify the active pocket, resulting in the optimal mutant H245S with 1.55-fold enhanced synthetic activity raising the DHAvD yield to 6.1%. This study achieved the first enzymatic synthesis of DHAvD and characterized its enzymatic properties. The work not only lays a foundation for subsequent development but also provides theoretical and technical insights for research on other lipases. ? 2026 Chinese Academy of Sciences. All rights reserved.