Multivariate modular metabolic engineering of Escherichia coli to produce resveratrol from l-tyrosine

Wu, Junjun; Liu, Peiran; Fan, Yongming; Bao, Han; Du, Guocheng; Zhou, Jingwen; Chen, Jian

doi:10.1016/j.jbiotec.2013.07.030

Cited by 116 publications

(99 citation statements)

References 23 publications

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“…Therefore, balancing relative gene expression is a potential route to the rational and efficient regulation of the expression of genes in the overall pathway. Our previous studies showed that coumaroyl-CoA can inhibit TAL enzyme activity, and the combination of pCDF-TAL-4CL and pET-CHS-CHI was shown to be the optimal combination for flavonoid biosynthesis (16,41). In this study, pACYC-tF3=H-tCPR was shown to be a better choice than pRSF-tF3=H-tCPR for optimization of the expression of tF3=H-tCPR.…”

Section: Discussionmentioning

confidence: 68%

Efficient Synthesis of Eriodictyol from l -Tyrosine in Escherichia coli

Zhu

et al. 2014

Appl Environ Microbiol

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The health benefits of flavonoids for humans are increasingly attracting attention. Because the extraction of high-purity flavonoids from plants presents a major obstacle, interest has emerged in biosynthesizing them using microbial hosts. Eriodictyol is a flavonoid with anti-inflammatory and antioxidant activities. Its efficient synthesis has been hampered by two factors: the poor expression of cytochrome P450 and the low intracellular malonyl coenzyme A (malonyl-CoA) concentration in Escherichia coli. To address these issues, a truncated plant P450 flavonoid, flavonoid 3=-hydroxylase (tF3=H), was functionally expressed as a fusion protein with a truncated P450 reductase (tCPR) in E. coli. This allowed the engineered E. coli to produce eriodictyol from L-tyrosine by simultaneously coexpressing the fusion protein with tyrosine ammonia lyase (TAL), 4-coumarate-CoA ligase (4CL), chalcone synthase (CHS), and chalcone isomerase (CHI). In addition, metabolic engineering was employed to enhance the availability of malonyl-CoA so as to achieve a new metabolic balance and rebalance the relative expression of genes to enhance eriodictyol accumulation. This approach made the production of eriodictyol 203% higher than that in the control strain. By using these strategies, the production of eriodictyol from L-tyrosine reached 107 mg/liter. The present work offers an approach to the efficient synthesis of other hydroxylated flavonoids from L-tyrosine or even glucose in E. coli.

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Section: Discussionmentioning

confidence: 68%

Efficient Synthesis of Eriodictyol from l -Tyrosine in Escherichia coli

Zhu

et al. 2014

Appl Environ Microbiol

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“…6). The presented strategy could be employed for the efficient production of many useful compounds that require malonyl-CoA as a precursor, such as the flavanones (2, 18), resveratrol (29), and polyketides (6). In E. coli, malonyl-CoA is consumed only for synthesizing fatty acids (6,7).…”

Section: Discussionmentioning

confidence: 99%

Fine-Tuning of the Fatty Acid Pathway by Synthetic Antisense RNA for Enhanced (2 S )-Naringenin Production from l -Tyrosine in Escherichia coli

et al. 2014

Appl Environ Microbiol

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cMalonyl coenzyme A (malonyl-CoA) is an important precursor for the synthesis of natural products, such as polyketides and flavonoids. The majority of this cofactor often is consumed for producing fatty acids and phospholipids, leaving only a small amount of cellular malonyl-CoA available for producing the target compound. The tuning of malonyl-CoA into heterologous pathways yields significant phenotypic effects, such as growth retardation and even cell death. In this study, fine-tuning of the fatty acid pathway in Escherichia coli with antisense RNA (asRNA) to balance the demands on malonyl-CoA for target-product synthesis and cell health was proposed. To establish an efficient asRNA system, the relationship between sequence and function for asRNA was explored. It was demonstrated that the gene-silencing effect of asRNA could be tuned by directing asRNA to different positions in the 5=-UTR (untranslated region) of the target gene. Based on this principle, the activity of asRNA was quantitatively tailored to balance the need for malonyl-CoA in cell growth and the production of the main flavonoid precursor, (2S)-naringenin. Appropriate inhibitory efficiency of the anti-fabB/fabF asRNA improved the production titer by 431% (391 mg/liter). Therefore, the strategy presented in this study provided a useful tool for the fine-tuning of endogenous gene expression in bacteria.

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“…In a study by Wu et al, resveratrol production was improved by multivariate modular optimization, which involved the following three modules: (i) module one consists of genes encoding TAL and 4CL for the production of p-coumaroyl-CoA from tyrosine; (ii) module two consists of genes encoding STS for resveratrol production from p-coumaroyl-CoA; (iii) module three consists of genes encoding MatB and MatC for increasing the intracellular malonylCoA pool. [132] The authors adjusted the expression level of each module using different promoter strengths (T7 or Trc) and gene copy numbers determined by origins of replication (pACYCDuet-1 harboring p15A origin, pCDFDuet-1 harboring CloDF13 (CDF) origin, pETDuet-1 harboring pBR322 origin, and pRSFDuet-1 harboring RSF 1030 (RSF) origin) in a previously reported manner. [104a] After comparing multiple combinations of promoter strengths and gene copy numbers, the optimum strain produced 35.02 mg L −1 of resveratrol from tyrosine.…”

Section: Modular Optimizationmentioning

confidence: 99%

“…[104a] After comparing multiple combinations of promoter strengths and gene copy numbers, the optimum strain produced 35.02 mg L −1 of resveratrol from tyrosine. [132] …”

Section: Modular Optimizationmentioning

confidence: 99%

Metabolic Engineering of Microorganisms for the Production of Natural Compounds

Park

Yang

et al. 2017

Adv. Biosys.

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Natural products have been attracting much interest around the world for their diverse applications, especially in drug and food industries. Plants have been a major source of many different natural products. However, plants are affected by weather and environmental conditions and their successful extraction is rather limited. Chemical synthesis is inefficient due to the complexity of their chemical structures involving enantioselectivity and regioselectivity. For these reasons, an alternative means of overproducing valuable natural products using microorganisms has emerged. In recent years, various metabolic engineering strategies have been developed for the production of natural products by microorganisms. Here, the strategies taken to produce natural products are reviewed. For convenience, natural products are classified into four main categories: terpenoids, phenylpropanoids, polyketides, and alkaloids. For each product category, the strategies for establishing and rewiring the metabolic network for heterologous natural product biosynthesis, systems approaches undertaken to optimize production hosts, and the strategies for fermentation optimization are reviewed. Taken together, metabolic engineering has enabled microorganisms to serve as a prominent platform for natural compounds production. This article examines both the conventional and novel strategies of metabolic engineering, providing general strategies for complex natural compound production through the development of robust microbial‐cell factories.

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Multivariate modular metabolic engineering of Escherichia coli to produce resveratrol from l-tyrosine

Cited by 116 publications

References 23 publications

Efficient Synthesis of Eriodictyol from l -Tyrosine in Escherichia coli

Efficient Synthesis of Eriodictyol from l -Tyrosine in Escherichia coli

Fine-Tuning of the Fatty Acid Pathway by Synthetic Antisense RNA for Enhanced (2 S )-Naringenin Production from l -Tyrosine in Escherichia coli

Metabolic Engineering of Microorganisms for the Production of Natural Compounds

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