Efficient conversion of acetate into phloroglucinol by recombinant Escherichia coli

Xu, Xin; Xian, Ming; Liu, Huizhou

doi:10.1039/c7ra09519h

Cited by 18 publications

(23 citation statements)

References 37 publications

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“…Microbial chemical production from acetate is an emerging eld and commonly relies on the use of complex media additives [17,18,20,21]. Therefore, it was surprising that the addition of yeast extract did not allow 2,3-butanediol and acetoin formation from acetate, but only from yeast extract.…”

Section: Discussionmentioning

confidence: 99%

“…Comparing this study to recent literature [14,15,17,18] shows that the major difference is the metabolic intermediate, from which the product is derived. While the precursor for other products such as acetone, isopropanol, 3-hydroxypropionic acid and phloroglucinol is acetyl-CoA [14,17,20,21], 2,3-butanediol is derived from pyruvate. Therefore, to produce 2,3-butanediol from acetate, acetate must rst be converted into pyruvate.…”

Section: Discussionmentioning

confidence: 99%

“…Overexpression of either of these pathways has been shown to increase acetate uptake and product formation [14,17,20,21]. Other strategies that resulted in increased product formation were the deletion of icdA, which led to blocking of the TCA cycle and increased acetone synthesis [21] or deletion of iclR, which activated the glyoxylate shunt and improved 3-hydroxypropionic acid production in combination with acs overexpression [14].…”

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confidence: 99%

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Microbial upgrading of acetate into 2,3-butanediol and acetoin by E. coli W

Novak

Kutscha

Pflügl

2020

Preprint

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Background: Acetate is an abundant carbon source and its use as an alternative feedstock has great potential for the production of fuel and platform chemicals. Acetoin and 2,3-butanediol represent two of these potential platform chemicals.Results: The aim of this study was to produce 2,3-butanediol and acetoin from acetate in Escherichia coli W. The key strategies to achieve this goal were: strain engineering, in detail the deletion of mixed acid fermentation pathways E. coli W ΔldhA ΔadhE Δpta ΔfrdA 445_Ediss and the development of a new defined medium containing five amino acids and seven vitamins. Stepwise reduction of the media additives further revealed that diol production from acetate is mediated by the availability of aspartate. Other amino acids or TCA cycle intermediates did not enable growth on acetate. Cultivation under controlled conditions in batch and pulsed fed-batch experiments showed that aspartate was consumed before acetate, indicating that co-utilization is not a prerequisite for diol production. The addition of aspartate gave cultures a start-kick and was not required for feeding. Pulsed fed-batches resulted in the production of 1.43 g l-1 from aspartate and acetate and 1.16 g l-1 diols (2,3-butanediol and acetoin) from acetate alone. The yield reached 0.09 g diols per g acetate, which accounts for 26 % of the theoretical maximum.Conclusion: This study for the first time showed acetoin and 2,3-butanediol production from acetate as well as the use of chemically defined medium for product formation from acetate in E. coli. Hereby, we provide a solid base for process intensification and the investigation of other potential products.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Microbial upgrading of acetate into 2,3-butanediol and acetoin by E. coli W

Novak

Kutscha

Pflügl

2020

Preprint

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“…Previously, we have accomplished the production of PG and mevalonate from acetate in recombinant E. coli with ACS overexpression [21,22]. In this study, we constructed an engineered E. coli strain growing with ethanol as sole carbon source by introduction of AdhE A267T/E568K mutant, and achieved the conversion of ethanol into various bioproducts including polyhydroxybutyrate (PHB), PG and 3HP.…”

Section: Introductionmentioning

confidence: 99%

Comparison of glucose, acetate and ethanol as carbon resource for production of acetyl-CoA derived chemicals in engineered Escherichia coli

Sun

Ding

Liu

et al. 2020

Preprint

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Background: Acetyl-CoA is a fundamental metabolite in Escherichia coli, and also a precursor for biosynthesis of chemicals and materials suitable for multiple applications. The acetyl-CoA synthesis route from glucose presents low atomic economy due to the release of CO2 in pyruvate decarboxylation reaction. Because ethanol and acetate, both ordinary and inexpensive chemicals, can be converted into acetyl-CoA directly, they could be alternative substrates for production of acetyl-CoA derivatives. Results: In this study, the bifunctional reductase AdhE mutant (A267T/E568K), which converts ethanol into acetyl-CoA, was used to enable E. coli to grow on ethanol, and AMP-forming acetyl-CoA synthetase ACS was employed to enhance the ability of E. coli to utilize acetate. Several products derived from acetyl-CoA, including polyhydroxybutyrate, 3-hydroxypropionate, and phloroglucinol, were produced from glucose, ethanol, and acetate, respectively, by engineered E. coli strains. Compared with glucose and acetate, the strains grown on ethanol presented the highest production and yield of carbon source, and metabolome analysis revealed the reasons of high yield from ethanol. Conclusions: The conversion of ethanol into acetyl-CoA presents high atomic economy along with generation of reducing power, and the yield of target chemical from ethanol is much higher than those from glucose and acetate. All these results suggested that ethanol could be a suitable carbon source for production of acetyl-CoA derived bioproducts.

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“…Acetate uptake in E. coli is possible via two routes: the high a nity, irreversible acetyl-CoA synthetase (acs) or the low a nity, reversible acetate kinase -phosphate acetyl transferase (ackA-pta) system [18]. Overexpression of either of these pathways has been shown to increase acetate uptake and product formation [13,16,19,20]. Other strategies that resulted in increased product formation were the deletion of icdA, which led to blocking of the TCA cycle and increased acetone synthesis [20] or deletion of iclR, which activated the glyoxylate shunt and improved 3-hydroxypropionic acid production in combination with acs overexpression [13].…”

Section: Introductionmentioning

confidence: 99%

Microbial Upgrading of Acetate into 2,3-Butanediol and Acetoin by E. coli W

Novak

Kutscha

Pflügl

2020

Preprint

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BackgroundAcetate is an abundant carbon source and its use as an alternative feedstock has great potential for the production of fuel and platform chemicals. Acetoin and 2,3-butanediol represent two of these potential platform chemicals. ResultsThe aim of this study was to produce 2,3-butanediol and acetoin from acetate in Escherichia coli W. The key strategies to achieve this goal were: strain engineering, in detail the deletion of mixed acid fermentation pathways E. coli W ΔldhA ΔadhE Δpta ΔfrdA 445_Ediss and the development of a new defined medium containing five amino acids and seven vitamins. Stepwise reduction of the media additives further revealed that diol production from acetate is mediated by the availability of aspartate. Other amino acids or TCA cycle intermediates did not enable growth on acetate. Cultivation under controlled conditions in batch and fed-batch experiments showed that aspartate was consumed before acetate, indicating that co-utilization is not a prerequisite for diol production. The addition of aspartate gave cultures a start-kick and was not required for feeding. Pulsed fed-batches resulted in the production of 1.43 g l-1 from aspartate and acetate and 1.16 g l-1 diols (2,3-butanediol and acetoin) from acetate alone. The yield reached 0.09 g diols per g acetate, which accounts for 26 % of the theoretical maximum.ConclusionThis study for the first time showed acetoin and 2,3-butanediol production from acetate as well as the use of chemically defined medium for product formation from acetate in E. coli. Hereby, we provide a solid base for process intensification and the investigation of other potential products.

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Efficient conversion of acetate into phloroglucinol by recombinant Escherichia coli

Abstract: Phloroglucinol, an important fine chemical, was attempted to be produced by a recombinant Escherichia coli, using acetate, a less costly feedstock, as a alternative carbon source.

Cited by 18 publications

References 37 publications

Microbial upgrading of acetate into 2,3-butanediol and acetoin by E. coli W

Microbial upgrading of acetate into 2,3-butanediol and acetoin by E. coli W

Comparison of glucose, acetate and ethanol as carbon resource for production of acetyl-CoA derived chemicals in engineered Escherichia coli

Microbial Upgrading of Acetate into 2,3-Butanediol and Acetoin by E. coli W

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