Stepwise modular pathway engineering of Escherichia coli for efficient one-step production of (2S)-pinocembrin

Wu, Junjun; Zhang, Xia; Dong, Mingsheng; Zhou, Jingwen

doi:10.1016/j.jbiotec.2016.06.007

Cited by 31 publications

(25 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Here, a rational modular design approach was developed for grouping and optimizing modules. Compared to our previous studies 14, 16, 17 , this rational modular design approach demonstrated that choosing separating node at cinnamic acid rather than previous cinnamoyl-CoA lead to a dramatic increase in final production titer. Final pinosylvin titers were improved to 281 mg/L, which represented the highest titer reported to date.…”

Section: Introductionmentioning

confidence: 69%

“…One was selected from the red yeast Rhodotorula glutinis (RgPAL), which was successfully used in our previous studies 16, 19 . The other one was chosen from Trichosporon cutaneum (TcPAL), which was a novel phenylalanine/tyrosine ammonia-lyases exhibiting high activities toward both L-phenylalanine and L-tyrosine 20 .…”

Section: Resultsmentioning

confidence: 99%

“…Based on our previous study, aroF wt and pheA fbr under T7 promoter in pCOLADuet-1 (pCOLA-aroF wt -pheA fbr ), RgPAL or TcPAL and 4CL under Trc promoter in pCDFDuet-1 (pCDFD-Trc-RgPAL-Trc-4CL or pCDFD-Trc-TcPAL-Trc-4CL), STS under T7 promoter in pETDuet-1 (pETD-STS) were overexpressed in E. coli BL21(DE3) strain to construct the initial fermentation platform, as this combination always led to the highest flavonoid 16, 21 or stilbene titer 14 . We found that the combination containing TcPAL produced higher pinosylvin (10.5 mg/L) than RgPAL (3.7 mg/L).…”

Section: Resultsmentioning

confidence: 99%

“…This genetic recalcitrance restricted the ability to balance the overall pathway. Previously, we demonstrated that reducing the 5′ region secondary structure of the open reading frame of target gene mRNA could function in improving protein expression with no additional metabolic burden 16 . Hence, in this study, this strategy was further developed to optimizing and balancing the heterologous gene expression coding for enzymes of the pinosylvin production pathway.…”

Section: Resultsmentioning

confidence: 99%

“…Based on our previous study 16 , changing synonymous codon usage in the first 14 amino acids of N-terminal protein was performed. For each codon, the first and second positions were kept constant while the third base position mutated randomly.…”

Section: Resultsmentioning

confidence: 99%

See 4 more Smart Citations

Rational modular design of metabolic network for efficient production of plant polyphenol pinosylvin

Zhang

Zhu

et al. 2017

Sci Rep

Self Cite

View full text Add to dashboard Cite

Efficient biosynthesis of the plant polyphenol pinosylvin, which has numerous applications in nutraceuticals and pharmaceuticals, is necessary to make biological production economically viable. To this end, an efficient Escherichia coli platform for pinosylvin production was developed via a rational modular design approach. Initially, different candidate pathway enzymes were screened to construct de novo pinosylvin pathway directly from D-glucose. A comparative analysis of pathway intermediate pools identified that this initial construct led to the intermediate cinnamic acid accumulation. The pinosylvin synthetic pathway was then divided into two new modules separated at cinnamic acid. Combinatorial optimization of transcriptional and translational levels of these two modules resulted in a 16-fold increase in pinosylvin titer. To further improve the concentration of the limiting precursor malonyl-CoA, the malonyl-CoA synthesis module based on clustered regularly interspaced short palindromic repeats interference was assembled and optimized with other two modules. The final pinosylvin titer was improved to 281 mg/L, which was the highest pinosylvin titer even directly from D-glucose without any additional precursor supplementation. The rational modular design approach described here could bolster our capabilities in synthetic biology for value-added chemical production.

show abstract

Section: Introductionmentioning

confidence: 69%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

See 3 more Smart Citations

Rational modular design of metabolic network for efficient production of plant polyphenol pinosylvin

Zhang

Zhu

et al. 2017

Sci Rep

Self Cite

View full text Add to dashboard Cite

show abstract

Microbial Metabolic Pathways in the Production of Valued‐added Products

Pereira

Finco

Letti

et al. 2018

Microbial Sensing in Fermentation

View full text Add to dashboard Cite

Expanding the Chemical Palette of Industrial Microbes: Metabolic Engineering for Type III PKS‐Derived Polyketides

Palmer

Alper

2018

Biotechnology Journal

View full text Add to dashboard Cite

Polyketides are a unique class of molecules with attractive bioactive and chemical properties. As a result, biorenewable production is being explored with these molecules as potential pharmaceutical, fuel, and material precursors. In particular, type III polyketide synthases enable access to a diverse class of chemicals using a relatively simple biochemical synthesis pathway. In this review, the recent advances in the engineering of microbial hosts for the production of type III PKS‐derived polyketides are highlighted. In particular, the field has moved beyond simple proof‐of‐concept and has been exploring engineering efforts that have led to improved production scales. This review details engineering progress for the production of acetyl‐CoA‐ and malonyl‐CoA‐derived polyketides including the products triacetic acid lactone and phloroglucinol as well as polyphenolic, phenylpropanoid‐derived compounds including flavonoids, stilbenoids, and curcuminoids. Specifically, the authors focus on enumerating the metabolic engineering strategies employed and product titers achieved for these molecules. Finally, the authors highlight tools and strategies that can be leveraged to realize the potential of microbial production and diversification of these molecules.

show abstract

Stepwise modular pathway engineering of Escherichia coli for efficient one-step production of (2S)-pinocembrin

Cited by 31 publications

References 40 publications

Rational modular design of metabolic network for efficient production of plant polyphenol pinosylvin

Rational modular design of metabolic network for efficient production of plant polyphenol pinosylvin

Microbial Metabolic Pathways in the Production of Valued‐added Products

Expanding the Chemical Palette of Industrial Microbes: Metabolic Engineering for Type III PKS‐Derived Polyketides

Contact Info

Product

Resources

About