文献类型：期刊文献

英文题名：Reductive Amination of Biobased Levulinic Acid to Unnatural Chiral γ-Amino Acid Using an Engineered Amine Dehydrogenase

作者：Cai, Rui-Feng[1];Liu, Lei[1];Chen, Fei-Fei[1];Li, Aitao[2,3];Xu, Jian-He[1,3];Zheng, Gao-Wei[1,3]

机构：[1]East China Univ Sci & Technol, Shanghai Collaborat Innovat Ctr Biomfg, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Hubei Univ, Sch Life Sci, State Key Lab Biocatalysis & Enzyme Engn, Hubei Key Lab Ind Biotechnol, 368 Youyi Rd, Wuhan 430062, Peoples R China;[3]East China Univ Sci & Technol, Frontiers Sci Ctr Materiobiol & Dynam Chem, Shanghai 200237, Peoples R China

年份：2020

卷号：8

期号：46

起止页码：17054

外文期刊名：ACS SUSTAINABLE CHEMISTRY & ENGINEERING

收录：;EI(收录号：20204809548664);WOS:【SCI-EXPANDED(收录号:WOS:000595489200007)】；

基金：The work was financially supported by the National Natural Science Foundation of China (nos. 21878085, 21536004, and 21871085), the National Key Research and Development Program of China (2019YFA09005000), the Fundamental Research Funds for the Central Universities (22221818014), and the Open Funding Project of the State Key Laboratory of Biocatalysis and Enzyme Engineering (SKLBEE2019001).

语种：英文

外文关键词：biomass; reductive amination; amine dehydrogenase; directed evolution; unnatural gamma-amino acid; (S)-4-aminopentanoic acid

摘要：Optically pure (S)-4-aminopentanoic acid is a pivotal precursor in the synthesis of therapeutic molecules and pyrrolidinone derivatives. Enantioselective reductive amination of levulinic acid catalyzed by amine dehydrogenases, a readily sustainable material from biobased lignocellulosic waste, represents an attractive approach for the synthesis of (S)-4-aminopentanoic acid. However, the natural amine dehydrogenases reported so far showed insufficient activity toward levulinic acid. Herein, we engineered a naturally occurring amine dehydrogenase from a thermophilic bacterium Petrotoga mobilis (PmAmDH) by directed evolution. The catalytic efficiency of the most active mutant PmAmDHI80T/P224S/E296G was elevated by 18 folds in comparison to the wild-type enzyme. Using PmAmDHI80T/P224S/E296G coupled with formate dehydrogenase for reduced nicotinamide adenine dinucleotide regeneration, 0.5 M of levulinic acid was reductively aminated in more than 97% conversion at 40 degrees C, generating the corresponding product (S)-4-aminopentanoic acid with >99% ee and 90% yield. Furthermore, we also successfully developed a chemoenzymatic cascade route for the synthesis of (S)-4-aminopentanoic acid from renewable starch. These results indicated that the engineered amine dehydrogenase PmAmDHI80T/P224S/E296G can serve as an efficient biocatalyst for the manufacture of highly valued chiral unnatural amino acids using renewable feedstocks.