Central pathway engineering for enhanced succinate biosynthesis from acetate in <i>Escherichia coli</i>

Huang, Bing; Yang, Hao; Fang, Guochen; Zhang, Xing; Wu, Hui; Li, Zhimin; Ye, Qin

doi:10.1002/bit.26528

Cited by 61 publications

(54 citation statements)

References 42 publications

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“…2 ). The activation of each molecule of acetate for acetyl-CoA synthesis in acs pathway requires two molecule of ATP, while pta - ackA pathway consumes only one molecule of ATP for acetyl-CoA synthesis [ 22 , 33 ]. It was reported also that the growth of E. coli stains on low concentrations of acetate depends on acs pathway, while growth on high concentrations requires pta - ackA pathway [ 28 , 29 ].…”

Section: Resultsmentioning

confidence: 99%

Metabolic engineering of Escherichia coli for the synthesis of polyhydroxyalkanoates using acetate as a main carbon source

Chen

Zhang

et al. 2018

Microb Cell Fact

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BackgroundHigh production cost of bioplastics polyhydroxyalkanoates (PHA) is a major obstacle to replace traditional petro-based plastics. To address the challenges, strategies towards upstream metabolic engineering and downstream fermentation optimizations have been continuously pursued. Given that the feedstocks especially carbon sources account up to a large portion of the production cost, it is of great importance to explore low cost substrates to manufacture PHA economically.ResultsEscherichia coli was metabolically engineered to synthesize poly-3-hydroxybutyrate (P3HB), poly(3-hydroxybutyrate-co-4-hydroxybutyrate) (P3HB4HB), and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) using acetate as a main carbon source. Overexpression of phosphotransacetylase/acetate kinase pathway was shown to be an effective strategy for improving acetate assimilation and biopolymer production. The recombinant strain overexpressing phosphotransacetylase/acetate kinase and P3HB synthesis operon produced 1.27 g/L P3HB when grown on minimal medium supplemented with 10 g/L yeast extract and 5 g/L acetate in shake flask cultures. Further introduction succinate semialdehyde dehydrogenase, 4-hydroxybutyrate dehydrogenase, and CoA transferase lead to the accumulation of P3HB4HB, reaching a titer of 1.71 g/L with a 4-hydroxybutyrate monomer content of 5.79 mol%. When 1 g/L of α-ketoglutarate or citrate was added to the medium, P3HB4HB titer increased to 1.99 and 2.15 g/L, respectively. To achieve PHBV synthesis, acetate and propionate were simultaneously supplied and propionyl-CoA transferase was overexpressed to provide 3-hydroxyvalerate precursor. The resulting strain produced 0.33 g/L PHBV with a 3-hydroxyvalerate monomer content of 6.58 mol%. Further overexpression of propionate permease improved PHBV titer and 3-hydroxyvalerate monomer content to 1.09 g/L and 10.37 mol%, respectively.ConclusionsThe application of acetate as carbon source for microbial fermentation could reduce the consumption of food and agro-based renewable bioresources for biorefineries. Our proposed metabolic engineering strategies illustrate the feasibility for producing polyhydroxyalkanoates using acetate as a main carbon source. Overall, as an abundant and renewable resource, acetate would be developed into a cost-effective feedstock to achieve low cost production of chemicals, materials, and biofuels.Electronic supplementary materialThe online version of this article (10.1186/s12934-018-0949-0) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

Metabolic engineering of Escherichia coli for the synthesis of polyhydroxyalkanoates using acetate as a main carbon source

Chen

Zhang

et al. 2018

Microb Cell Fact

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“…The second one is diverting AA towards SA production. The AA produced, an undesirable product, can be combined with xylose utilization for SA production and this co-fermentation will be bene cial for e cient utilization of lignocellulosic hydrolysate containing substantial amount of AA [34][35][36]. In order to understand the robustness of the recombinant Y. lipolytica PSA02004PP strain to withstand adverse condition such as low pH, the pH was not controlled in fermentations carried out in this study.…”

Section: Discussionmentioning

confidence: 99%

Bioproduction of succinic acid from xylose by engineered Yarrowia lipolytica without pH control

Prabhu

Amaro

Lin

et al. 2020

Preprint

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Background : Xylose is a most prevalent sugar available in hemicellulose fraction of lignocellulosic biomass (LCB) and of great interest for the green economy. Unfortunately, most of the cell factories cannot inherently metabolize xylose as sole carbon source. Yarrowia lipolytica is a non-conventional yeast to produce industrially important metabolites. The yeast is able to metabolize a large variety of substrates including both hydrophilic and hydrophobic carbon sources. However, Y. lipolytica lacks effective metabolic pathway for xylose uptake and only scarce information is available on utilization of xylose. For the economically feasibility of LCB-based biorefineries, effective utilization of both pentose and hexose sugars is obligatory. Results : In the present study, succinic acid (SA) production from xylose by Y. lipolytica was examined. To this end, Y. lipolytica PSA02004 strain was engineered by overexpressing pentose pathway cassette comprising of xylose reductase ( XR ), xylitol dehydrogenase ( XDH ) and xylulose kinase ( XK ) gene. The recombinant strain exhibited a robust growth on xylose as sole carbon source and produced substantial amount of SA. The inhibition of cell growth and SA formation was observed above 60 g/L xylose concentration. The batch cultivation of recombinant strain in bioreactor resulted in a maximum biomass concentration of 7.3 g/L and SA titer of 11.2 g/L with the yield of 0.19 g/g. Similar results in term of cell growth and SA production were obtained with xylose-rich hydrolysate derived from sugarcane bagasse. The fed-batch fermentation yielded biomass concentration of 11.8 g/L (OD 600 : 56.1) and SA titer of 22.3 g/L with a gradual decrease in pH below 4.0. Acetic acid was obtained as a main byproduct in all the fermentations. Conclusion : The recombinant strain displayed potential for bioconversion of xylose to SA. Further, this study provided a new insight on conversion of lignocellulosic biomass into value-added products. To the best of our knowledge, this is the first study on SA production by Y. lipolytica using xylose as a sole carbon source.

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“…Then, glucose can be fermented in the following anaerobic stage and 60.1 g L −1 of succinic acid was obtained with 1.00 g L −1 h −1 productivity . Optimization was carried out to enhance the acetic acid assimilation pathway, to increase the aerobic Adenosine triphosphate (ATP) supply, reduce OAA decarboxylation, and to engineer the TCA cycle; 3.6 g L −1 of succinic acid was accumulated directly from acetic acid, which reached the maximum theoretical yield …”

Section: Succinic Acid Production By Metabolically Engineered Strainsmentioning

confidence: 99%

Bio‐based succinic acid: an overview of strain development, substrate utilization, and downstream purification

Dai

Guo

Zhang

et al. 2019

Biofuels Bioprod Bioref

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Succinic acid is an industrially important platform chemical that could be used as precursor for the establishment of a sustainable chemical industry. Due to the depletion of crude oil and the need for sustainable development, the biological production of succinic acid from renewable resources has attracted wide attention. To establish an economical bio‐based succinic acid production process, various metabolic engineering strategies have been developed and applied to improve the strain performance, including efficient CO2 fixation, by‐product elimination, use of metabolic modeling approaches, engineering of transcriptional regulations and optimization of reducing power balance. The utilization of renewable waste resources and the optimization of purification process have also been investigated to decrease the cost. This review will provide insights for current advances in succinic acid production and perspectives on further research to make bio‐based succinic acid production more efficient and economical. © 2019 Society of Chemical Industry and John Wiley & Sons, Ltd

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Central pathway engineering for enhanced succinate biosynthesis from acetate in Escherichia coli

Cited by 61 publications

References 42 publications

Metabolic engineering of Escherichia coli for the synthesis of polyhydroxyalkanoates using acetate as a main carbon source

Metabolic engineering of Escherichia coli for the synthesis of polyhydroxyalkanoates using acetate as a main carbon source

Bioproduction of succinic acid from xylose by engineered Yarrowia lipolytica without pH control

Bio‐based succinic acid: an overview of strain development, substrate utilization, and downstream purification

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