期刊
ACS SYNTHETIC BIOLOGY
卷 6, 期 9, 页码 1784-1792出版社
AMER CHEMICAL SOC
DOI: 10.1021/acssynbio.7b00129
关键词
vanillyl alcohol; aromatic compounds; shikimate pathway; caffeate O-methyltransferase; enzyme promiscuity; microbial synthesis
资金
- National Natural Science Foundation of China [21406010, 21606012, 21636001]
- Programme of Introducing Talents of Discipline to Universities [B13005]
- Program for Changjiang Scholars and Innovative Research Team in Universities in China [IRT13045]
- Academic Leader of Beijing Polytechnic [DTR201601]
- Beijing Polytechnic [YZK028]
- College of Engineering
- University of Georgia, Athens
- Beijing University of Chemical Technology
Vanillyl alcohol is a phenolic alcohol and is used as a flavoring agent in foods and beverages. In this paper, we propose a novel artificial pathway for microbial production of vanillyl alcohol from simple carbon sources. The pathway extends from 4-hydroxybenzoic acid (4-HBA), and needs only three heterologous enzymes, p-hydroxybenzoate hydroxylase (PobA), carboxylic acid reductase (CAR) and caffeate O-methyltransferase (COMT). First, we examined the promiscuous activity of COMT toward 3,4-dihydroxybenzyl alcohol and found a k(cat) value of 0.097 s(-1). Meanwhile, 499.36 mg/L vanillyl alcohol was produced by COMT in vivo catalysis when fed with 1000 mg/L 3,4-dihydroxybenzyl alcohol. In the following experiment, de novo biosynthesis of vanillyl alcohol was carried out and 240.69 mg/L vanillyl alcohol was produced via modular optimization of pathway genes. This work was to date the first achievement for microbial production of vanillyl alcohol. Additionally, the present study demonstrates the application of enzyme promiscuity of COMT in the design of an artificial pathway for the production of high-value methylated aromatic compounds.
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