Pathway construction and metabolic engineering for fermentative production of β-alanine in Escherichia coli

Zou, Xinyu; Guo, Laixian; Huang, Lilong; Li, Miao; Zhang, Sheng; Yang, Anren; Zhang, Yu; Zhu, Lei; Zhang, Hongxia; Zhang, Juan; Feng, Zhibin

doi:10.1007/s00253-020-10359-8

Cited by 40 publications

(38 citation statements)

References 52 publications

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“…When pFT28-ectABC-EclysC*-aspC was transferred into MWZ004, the resulting strain MWZ004/pFT28-ectABC-EclysC*-aspC produced almost the same amount of ectoine as MWZ003/pFT28-ectABC-EclysC* (data not shown). Therefore, overexpression of aspC did not significantly increase the production of ectoine, which was consistent with previously reported studies on other aspartic acid-derived products [25,26].…”

Section: Introduce a Novel Aspartate Dehydrogenase From P Aeruginosa Pao1 To Promote Ectoine Synthesis In E Colisupporting

confidence: 92%

“…As previously reported, aspartate dehydrogenase encoded by aspDH from Pseudomonas aeruginosa PAO1 can aminate oxaloacetate directly to form aspartate [28]. This enzyme has been heterologously introduced to E. coli to enhance the accumulation of aspartate and promote the production of β-alanine [26].…”

Section: Introductionmentioning

confidence: 93%

“…The enzymatic activity deficiency of the aspartate kinase caused by thrA deletion can be compensated by overexpression of feedback-resistant lysC, which also encodes aspartokinase [20]. It is well documented that phosphoenolpyruvate carboxylase encoded by the gene ppc is the key enzyme catalyzing the biosynthesis of oxaloacetate [26], and the expression of ppc has an optimal level for l-threonine production [27]. As previously reported, aspartate dehydrogenase encoded by aspDH from Pseudomonas aeruginosa PAO1 can aminate oxaloacetate directly to form aspartate [28].…”

Section: Introductionmentioning

confidence: 99%

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Metabolic engineering of Escherichia coli for efficient ectoine production

Zhang

Fang

Zhu

et al. 2021

Syst Microbiol and Biomanuf

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Ectoine is a high-value stabilizer and protective agent with various applications in enzyme industry, cosmetics, and biomedicine. In this study, rational engineering strategies have been implemented in Escherichia coli to efficiently produce ectoine. First, the synthetic pathway of ectoine was constructed in E. coli MG1655 by introducing an artificial thermal switch system harboring the ectABC cluster from Halomonas elongate, and the resulting strain produced 1.95 g/L ectoine. Second, crr encoding the glucose-specific enzyme II domain A of phosphotransferase system and iclR encoding the glyoxylate shunt transcriptional repressor were deleted in E. coli for enhancing the oxaloacetate supply, leading to the increasement of the ectoine titer to 9.09 g/L. Third, thrA encoding the bifunctional aspartokinase/homoserine dehydrogenase was removed from the genome to weaken the competitive pathway; simultaneously, an endogenous feedback-resistant lysC was overexpressed to complement the enzymatic activity deficiency of the aspartate kinase, leading to 30.36% increase of ectoine titer. Next, the expression of phosphoenolpyruvate carboxylase was modulated with varying gradient strength promoters to accelerate the biosynthesis efficiency of ectoine. Finally, aspDH encoding aspartate dehydrogenase from Pseudomonas aeruginosa PAO1 was overexpressed to further improve the biosynthesis of ectoine. The final strain MWZ003/pFT28-ectABC-EclysC*-aspDH-ppc3 produced 30.37 g/L ectoine after 36-h fed-batch fermentation with a yield of 0.132 g/g glucose and a productivity of 0.844 g/(L h).

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Section: Introduce a Novel Aspartate Dehydrogenase From P Aeruginosa Pao1 To Promote Ectoine Synthesis In E Colisupporting

confidence: 92%

Section: Introductionmentioning

confidence: 93%

Section: Introductionmentioning

confidence: 99%

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Metabolic engineering of Escherichia coli for efficient ectoine production

Zhang

Fang

Zhu

et al. 2021

Syst Microbiol and Biomanuf

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“…Simple overexpression of aspartate decarboxylase and lactate dehydrogenases in R. opacus allowed us to demonstrate the production of beta-alanine and lactic acid from CO 2 but economically viable production titers would clearly require more elaborate metabolic engineering efforts. As an example, in Escherichia coli , efficient production of beta-alanine from glucose requires overexpression of all genes encoding enzymes of the reductive branch of TCA cycle and deletion of several pathways for side products (Piao et al 2019; Zou et al 2020). In E. coli , the uptake of beta-alanine is performed by an active amino acid transporter (Schneider et al 2004).…”

Section: Discussionmentioning

confidence: 99%

Production of biopolymer precursors beta-alanine and L-lactic acid from CO₂with metabolically versatileRhodococcus opacusDSM 43205

Salusjärvi

Ojala

Gopalacharyulu

et al. 2020

Preprint

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Hydrogen oxidizing autotrophic bacteria are promising hosts for CO2 conversion into chemicals. In this work, we engineered the metabolically versatile lithoautotrophic bacterium Rhodococcus opacus strain DSM 43205 for synthesis of polymer precursors. Aspartate decarboxylase (panD) or lactate dehydrogenase (ldh) were expressed for beta-alanine or L-lactic acid production, respectively. The heterotrophic cultivations on glucose produced 25 mg L-1 beta-alanine and 742 mg L-1 L-lactic acid, while autotrophic cultivations with CO2, H2 and O2 resulted in the production of 1.8 mg L-1 beta-alanine and 146 mg L-1 L-lactic acid. Beta-alanine was also produced at 345 µg L-1 from CO2 in electrobioreactors, where H2 and O2 were provided by water electrolysis. This work demonstrates that R. opacus DSM 43205 can be readily engineered to produce chemicals from CO2 and provides base for its further metabolic engineering.

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“…Large amounts of high-value-added products have been successfully produced by engineered microbes. As for β-alanine biosynthesis, two approaches belonging to synthetic biology are mainly applied: whole-cell biocatalysts [5] and metabolic engineering [6]. The method of whole-cell biocatalysts emphasizes the reaction from L-aspartate to β-alanine, so this method relies on the addition of L-aspartate, which leads to a higher cost and higher yield [5].…”

Section: Introductionmentioning

confidence: 99%

Enhancement of β-Alanine Biosynthesis in Escherichia coli Based on Multivariate Modular Metabolic Engineering

Zhou

2021

Biology

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β-alanine is widely used as an intermediate in industrial production. However, the low production of microbial cell factories limits its further application. Here, to improve the biosynthesis production of β-alanine in Escherichia coli, multivariate modular metabolic engineering was recruited to manipulate the β-alanine biosynthesis pathway through keeping the balance of metabolic flux among the whole metabolic network. The β-alanine biosynthesis pathway was separated into three modules: the β-alanine biosynthesis module, TCA module, and glycolysis module. Global regulation was performed throughout the entire β-alanine biosynthesis pathway rationally and systematically by optimizing metabolic flux, overcoming metabolic bottlenecks and weakening branch pathways. As a result, metabolic flux was channeled in the direction of β-alanine biosynthesis without huge metabolic burden, and 37.9 g/L β-alanine was generated by engineered Escherichia coli strain B0016-07 in fed-batch fermentation. This study was meaningful to the synthetic biology of β-alanine industrial production.

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Pathway construction and metabolic engineering for fermentative production of β-alanine in Escherichia coli

Cited by 40 publications

References 52 publications

Metabolic engineering of Escherichia coli for efficient ectoine production

Metabolic engineering of Escherichia coli for efficient ectoine production

Production of biopolymer precursors beta-alanine and L-lactic acid from CO₂with metabolically versatileRhodococcus opacusDSM 43205

Enhancement of β-Alanine Biosynthesis in Escherichia coli Based on Multivariate Modular Metabolic Engineering

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Pathway construction and metabolic engineering for fermentative production of β-alanine in Escherichia coli

Cited by 40 publications

References 52 publications

Metabolic engineering of Escherichia coli for efficient ectoine production

Metabolic engineering of Escherichia coli for efficient ectoine production

Production of biopolymer precursors beta-alanine and L-lactic acid from CO2with metabolically versatileRhodococcus opacusDSM 43205

Enhancement of β-Alanine Biosynthesis in Escherichia coli Based on Multivariate Modular Metabolic Engineering

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Product

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Production of biopolymer precursors beta-alanine and L-lactic acid from CO₂with metabolically versatileRhodococcus opacusDSM 43205