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Abstract
Phenylacetic acid (PAA) is a fine chemical with a high industrial demand for its widespread uses. Whereas, microorganic synthesis of PAA is impeded by the formation of by-product phenethyl alcohol due to quick, endogenous, and superfluous conversion of aldehydes to their corresponding alcohols, which resulted in less conversation of PAA from aldehydes. In this study, an Escherichia coli K-12 MG1655 strain with reduced aromatic aldehyde reduction (RARE) that does duty for a platform for aromatic aldehyde biosynthesis was used to prompt more PAA biosynthesis. We establish a microbial biosynthetic pathway for PAA production from the simple substrate phenylalanine in E. coli with heterologous coexpression of aminotransferase (ARO8), keto acid decarboxylase (KDC) and aldehyde dehydrogenase H (AldH) gene. It was found that PAA transformation yield was up to ~94% from phenylalanine in E. coli and there was no by-product phenethyl alcohol was detected. Our results reveal the high efficiency of the RARE strain for production of PAA and indicate the potential industrial applicability of this microbial platform for PAA biosynthesis.
Highlights
Phenylacetic acid (PAA) has received much attention on account of its extensive applications, which offer the huge demand
The phenylacetaldehyde dehydrogenase (FeaB, GenBank: 945933), aldehyde dehydrogenase B (AldB, GenBank: 948104) and aldehyde dehydrogenase H (AldH, GenBank: 8183735) gene were individually amplified by PCR from E. coli BL21 genomic DNA using primers FeaB-XbaI/FeaB-NheI–BamHI, AldB-XbaI/ AldB-SacI–BamHI and AldH-XbaI/AldH-NheI–BamHI separately, and individually inserted into pET28a(+) to give pDG3, pDG4 and pDG5
Construction of PAA biosynthetic pathway from glucose in E. coli The precursor substrate phenylpyruvate is an intermediates of shikimate pathway in E. coli
Summary
Phenylacetic acid (PAA) has received much attention on account of its extensive applications, which offer the huge demand. It has lots of applicable uses in medicine, pesticides, disinfectants and other industries (Dongamanti et al 2012; Duan et al 2000; Huang et al 2014a), and has been investigated as a kind of industrial raw material. Nowadays it is procured mainly by chemical methods. There are some strategies using enzyme catalytic synthesis of PAA, for example
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