Discovery of 2-Naphthoic Acid Monooxygenases by Genome Mining and their Use as Biocatalysts
Discovery of 2-Naphthoic Acid Monooxygenases by Genome Mining and their Use as Biocatalysts
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DOI：
10.1002/cssc.200900054
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发表时间：
2009-01-01
期刊：
影响因子：
8.4
通讯作者：
Kino, Kuniki
中科院分区：
文献类型：
--
作者：
Furuya, Toshiki;Kino, Kuniki
The large pool of cytochrome P450 (P450) open-reading frames identified in genome sequences has attracted much attention as a resource for new oxidation biocatalysts. P450 genes were cloned from genome-sequenced bacteria and coexpressed with putidaredoxin and its reductase genes to provide the redox partners of P450 in Escherichia coli. Whole-cell assays were performed with 2-napthoic acid as a substrate. Hydroxylated naphthoic acid products were rapidly detected with two reagents showing different colors in the presence of the products. Two P450s, CYP199A1 and CYP199A2 were found to hydroxylate the substrate to 7- and 8-hydroxy-2-naphthoic acids. The CYP199A1 whole-cell biocatalyst converted 1 mM 2-napthoic acid to 0.27 mM 7-hydroxy-2-naphthoic acid and 0.53 mM 8-hydroxy-2-naphthoic acid CYP199A2 exhibited similar regioselectivity to CYP199A1. Furthermore, we found that 8-hydroxy-2-naphthoic acid emits near-white fluorescence when exposed to UV light. These P450s will provide a facile and environmentally friendly synthetic approach to the hydroxynaphthoic acids.
