|
type |
Journal Article |
authors |
Chen M, Liu C-T, Tang Y |
title |
Discovery and Biocatalytic Application of a PLP-Dependent Amino Acid γ-Substitution Enzyme that Catalyzes C-C Bond Formation |
journal |
J Am Chem Soc |
Activity |
cndf |
Family |
cndf |
sel |
selected |
ui |
32434326 |
year |
(2020) |
volume |
142 |
number |
23 |
pages |
10506-10515 |
| |
keywords |
DOI: 10.1021/jacs.0c03535 |
abstract |
Pyridoxal phosphate (PLP)-dependent enzymes can catalyze transformations of L-amino acids at α, β and γ positions. These enzymes are frequently involved in the biosynthesis of nonproteinogenic amino acids as building blocks of natural products, and are attractive biocatalysts. Here, we report the discovery of a two-step enzymatic synthesis of (2S, 6S)-6-methyl pipecolate 1, from the biosynthetic pathway of citrinadin. The key enzyme CndF is PLP-dependent and catalyzes synthesis of (S)-2-amino-6-oxoheptanoate 3 that is in equilibrium with the cyclic Schiff base. The second enzyme CndE is a stereoselective imine reductase that gives 1. Biochemical characterization of CndF showed this enzyme performs γ-elimination of O-acetyl-L-homoserine to generate the vinylglycine ketimine, which is subjected to nucleophilic attack by acetoacetate to form the new Cγ-Cδ bond in 3 and complete the γ-substitution reaction. CndF displays promiscuity towards different β-keto carboxylate and esters. Using an Aspergillus strain expressing CndF and CndE, feeding various alkyl-β-keto esters led to the biosynthesis of 6-substituted L-pipecolates. The discovery of CndF expands the repertoire of reactions that can be catalyzed by PLP-dependent enzymes. |
fulltext |
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last changed |
2020/06/16 10:12 |
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