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Biosynthesis of (R)-(+)-perillyl alcohol by Escherichia coli expressing neryl pyrophosphate synthase.

ABSTRACT: (R)-(+)-perillyl alcohol is widely used in agricultural and anticarcinogenic fields. Microbial production of (R)-(+)-perillyl alcohol was investigated in this study. We optimized biosynthesis of (R)-(+)-perillyl alcohol in Escherichia coli by using neryl pyrophosphate synthase and NADPH regeneration. Engineering neryl pyrophosphate (NPP)-supplied pathway resulted in a 4-fold improvement of (R)-(+)-perillyl alcohol titer. Subsequently, combined engineering of p-cymene monooxygenase (CymA) expression and module for NADPH regeneration exhibited a 15.4-fold increase of titer over the initial strain S02. Finally, 453 mg/L (R)-(+)-perillyl alcohol was achieved in fed-batch fermentation, which is the highest (R)-(+)-perillyl alcohol titer in E. coli.

SUBMITTER: Sun C

PROVIDER: S-EPMC9077825 | biostudies-literature | 2022 May

REPOSITORIES: biostudies-literature

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Biosynthesis of (R)-(+)-perillyl alcohol by Escherichia coli expressing neryl pyrophosphate synthase.

Sun Chao C Zhang Rubing R Xie Congxia C

Engineering in life sciences 20220213 5

(R)-(+)-perillyl alcohol is widely used in agricultural and anticarcinogenic fields. Microbial production of (R)-(+)-perillyl alcohol was investigated in this study. We optimized biosynthesis of (R)-(+)-perillyl alcohol in Escherichia coli by using neryl pyrophosphate synthase and NADPH regeneration. Engineering neryl pyrophosphate (NPP)-supplied pathway resulted in a 4-fold improvement of (R)-(+)-perillyl alcohol titer. Subsequently, combined engineering of p- ...[more]

PMID: 35573132

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Effectiveness of recombinant Escherichia coli on the production of (R)-(+)-perillyl alcohol.

Project description:Background(R)-(+)-perillyl alcohol is a naturally oxygenated monoterpene widely used as the natural flavor additives, insecticides, jet fuels and anti-cancer therapies. It was also readily available monoterpene precursors. However, this natural product is present at low concentrations from plant sources which are not economically viable. Therefore, alternative microbial production methods are rapidly emerging as an attractive alternative to make (R)-(+)-perillyl alcohol production more sustainable and environmentally friendly.ResultsWe engineered Escherichia coli to possess a heterologous mevalonate (MVA) pathway, including limonene synthase, P-cymene monoxygenase hydroxylase and P-cymene monoxygenase reductase for the production of (R)-(+)-perillyl alcohol. The concentration of (R)-(+)-limonene (the monoterpene precursor to (R)-(+)-perillyl alcohol) reached 45 mg/L from glucose. Enhanced (R)-(+)-perillyl alcohol production was therefore achieved. The strain produced (R)-(+)-perillyl alcohol at a titer of 87 mg/L and a yield of 1.5 mg/g glucose in a 5 L bioreactor fed batch system.ConclusionsThese datas highlight the efficient production of (R)-(+)-perillyl alcohol through the mevalonate pathway from glucose. This method serves as a platform for the future production of other monoterpenes.

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Preliminary X-ray diffraction analysis of octaprenyl pyrophosphate synthase from Escherichia coli.

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Enhancing epi-cedrol production in Escherichia coli by fusion expression of farnesyl pyrophosphate synthase and epi-cedrol synthase.

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Efficient Synthesis of (R)-(+)-Perillyl Alcohol From (R)-(+)-Limonene Using Engineered Escherichia coli Whole Cell Biocatalyst.

Project description:(R)-(+)-perillyl alcohol is a much valued supplemental compound with a wide range of agricultural and pharmacological characteristics. The aim of this study was to improve (R)-(+)-perillyl alcohol production using a whole-cell catalytic formula. In this study, we employed plasmids with varying copy numbers to identify an appropriate strain, strain 03. We demonstrated that low levels of alKL provided maximal biocatalyst stability. Upon determination of the optimal conditions, the (R)-(+)-perillyl alcohol yield reached 130 mg/L. For cofactor regeneration, we constructed strain 10, expressing FDH from Candida boidinii, and achieved (R)-(+)-perillyl alcohol production of 230 mg/L. As a result, 1.23 g/L (R)-(+)-perillyl alcohol was transformed in a 5 L fermenter. Our proposed method facilitates an alternative approach to the economical biosynthesis of (R)-(+)-perillyl alcohol.

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