期刊
ACS CATALYSIS
卷 12, 期 17, 页码 10700-10710出版社
AMER CHEMICAL SOC
DOI: 10.1021/acscatal.2c02369
关键词
biocatalysis; multi-enzyme cascade; non-standard amino acid; pyridoxal phosphate; Meinwald rearrangement; mechanism
资金
- Office of the Vice Chancellor for Research and Graduate Education at the University of Wisconsin -Madison
- Wisconsin Alumni Research Foundation
- NIH [DP2-GM137417, T32- GM135066, T32-GM008349]
- American Chemical Society Petroleum Research Fund
- National Science Foundation (NSF- MRI) [CHE-1429616]
Multi-enzyme biocatalytic cascades provide a practical route for the synthesis of complex bioactive molecules. In this study, a chemoenzymatic platform was developed to convert aryl epoxides into alpha-aryl aldehydes and then further react with L-threonine to produce beta-hydroxy-alpha-amino acids. The whole-cell biocatalyst, produced through co-expression of two enzymes in E. coli, demonstrated the ability to synthesize a variety of stereopure non-standard amino acids on a gram scale.
Multi-enzyme biocatalytic cascades are emerging as practical routes for the synthesis of complex bioactive molecules. However, the relative sparsity of water-stable carbon electrophiles limits the synthetic complexity of molecules made from such cascades. Here, we develop a chemoenzymatic platform that leverages styrene oxide isomerase (SOI) to convert readily accessible aryl epoxides into alpha-aryl aldehydes through Meinwald rearrangement. These unstable aldehyde intermediates are then intercepted with a C-C bond forming enzyme, ObiH, that catalyzes a transaldolase reaction with L-threonine to yield synthetically challenging beta-hydroxy-alpha-amino acids. Co-expression of both enzymes in E. coli yields a whole-cell biocatalyst capable of synthesizing a variety of stereopure non-standard amino acids (nsAA) and can be produced on a gram scale. We used isotopically labeled substrates to probe the mechanism of SOI, which we show to catalyze a concerted isomerization featuring a stereospecific 1,2-hydride shift. The viability of in situ generated alpha-aryl aldehydes was further established by intercepting them with a recently engineered decarboxylative aldolase to yield gamma-hydroxy nsAAs. Together, these data establish a versatile method of producing alpha-aryl aldehydes in simple, whole -cell conditions and show that these intermediates are useful synthons in C-C bond forming cascades.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据