Increasing l-threonine production in Escherichia coli by engineering the glyoxylate shunt and the l-threonine biosynthesis pathway

Zhao, Hui; Fang, Yu; Wang, Xiaoyuan; Zhao, Lei; Wang, Jianli; Li, Ye

doi:10.1007/s00253-018-9024-3

Cited by 48 publications

(58 citation statements)

References 43 publications

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“…Genome sequence of E. coli MG1655 was used to design the primer sequences because the strain TWF001 [9] was derived from MG1655. The thr operon regulatory region was amplified using the genome sequence of MG1655 as template because the sequence of thr operon regulatory region in TWF001 was mutated.…”

Section: Bacterial Growth Conditionmentioning

confidence: 99%

“…In previous work, we have developed an l-threonineproducing strain TWF003 from E. coli TWF001 by deleting iclR. The l-threonine production in TWF003 reached 11.76 g/L, which is a 26% increase compared to TWF001 [9]. Deleting fadR [6] or arcA [46] in TWF003 could further increase l-threonine production.…”

Section: Expression Regulation Of Iclr To Increase L-threonine Producmentioning

confidence: 99%

“…Escherichia coli is an important strain for l-threonine production [2][3][4]. Efforts have been made to improve the productivity of l-threonine in E. coli, such as increasing the carbon flux to l-threonine [5,6], enhancing the export of l-threonine [7], weakening the competitive pathway and reducing the consumption of l-threonine [8,9].…”

Section: Introductionmentioning

confidence: 99%

“…From the 6th codons of this leader peptide, it starts to encode l-threonine residues, therefore, it can regulate the transcription of the thr operon through the availability of l-threonine, by forming different base-paired RNA structure [11]. Since thrL inhibits l-threonine accumulation it has been mutated in l-threonine producing E. coli, such as TWF001 [9]. On the other hand, some biosynthetic pathways in E. coli can be activated in the presence of high concentration of l-threonine, such as the sulfate metabolism branch of the cysteine biosynthetic pathway which includes the genes cysDN, cysJI and cysH [12].…”

Section: Introductionmentioning

confidence: 99%

“…Based on this principle, various biosensors for amino acids and their precursors, including l-methionine, l-leucine, l-isoleucine, l-valine [13,14], l-lysine, l-arginine, l-serine, O-acetyl-l-serine [15], O-acetyl homoserine [16], and oxygen [17] have been constructed, and some of these sensors have been successfully applied to the high-throughput screening [12,18,19] and systems metabolic engineering [20]. l-Threonine production in E. coli can be enhanced by regulating IclR [8,9], a transcription factor. IclR is a repressor for the aceBAK operon ( Fig.…”

Section: Introductionmentioning

confidence: 99%

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Expression regulation of multiple key genes to improve l-threonine in Escherichia coli

Zhao

Yang

et al. 2020

Microb Cell Fact

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Background: Escherichia coli is an important strain for l-threonine production. Genetic switch is a ubiquitous regulatory tool for gene expression in prokaryotic cells. To sense and regulate intracellular or extracellular chemicals, bacteria evolve a variety of transcription factors. The key enzymes required for l-threonine biosynthesis in E. coli are encoded by the thr operon. The thr operon could coordinate expression of these genes when l-threonine is in short supply in the cell. Results: The thrL leader regulatory elements were applied to regulate the expression of genes iclR, arcA, cpxR, gadE, fadR and pykF, while the threonine-activating promoters P cysH , P cysJ and P cysD were applied to regulate the expression of gene aspC, resulting in the increase of l-threonine production in an l-threonine producing E. coli strain TWF001. Firstly, different parts of the regulator thrL were inserted in the iclR regulator region in TWF001, and the best resulting strain TWF063 produced 16.34 g l-threonine from 40 g glucose after 30 h cultivation. Secondly, the gene aspC following different threonine-activating promoters was inserted into the chromosome of TWF063, and the best resulting strain TWF066 produced 17.56 g l-threonine from 40 g glucose after 30 h cultivation. Thirdly, the effect of expression regulation of arcA, cpxR, gadE, pykF and fadR was individually investigated on l-threonine production in TWF001. Finally, using TWF066 as the starting strain, the expression of genes arcA, cpxR, gadE, pykF and fadR was regulated individually or in combination to obtain the best strain for l-threonine production. The resulting strain TWF083, in which the expression of seven genes (iclR, aspC, arcA, cpxR, gadE, pykF, fadR and aspC) was regulated, produced 18.76 g l-threonine from 30 g glucose, 26.50 g l-threonine from 40 g glucose, or 26.93 g l-threonine from 50 g glucose after 30 h cultivation. In 48 h fed-batch fermentation, TWF083 could produce 116.62 g/L l-threonine with a yield of 0.486 g/g glucose and productivity of 2.43 g/L/h. Conclusion: The genetic engineering through the expression regulation of key genes is a better strategy than simple deletion of these genes to improve l-threonine production in E. coli. This strategy has little effect on the intracellular metabolism in the early stage of the growth but could increase l-threonine biosynthesis in the late stage.

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Section: Bacterial Growth Conditionmentioning

confidence: 99%

Section: Expression Regulation Of Iclr To Increase L-threonine Producmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Expression regulation of multiple key genes to improve l-threonine in Escherichia coli

Zhao

Yang

et al. 2020

Microb Cell Fact

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Metabolic Engineering of Escherichia coli

Luo

Ahn

Chae

et al. 2021

Metabolic Engineering

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Bioretrosynthesis of Functionalized N‐Heterocycles from Glucose via One‐Pot Tandem Collaborations of Designed Microbes

Feng

Zhang

et al. 2020

Advanced Science

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The design of multistrain systems has markedly expanded the prospects of using long biosynthetic pathways to produce natural compounds. However, the cooperative use of artificially engineered microbes to synthesize xenobiotic chemicals from renewable carbohydrates is still in its infancy. Here, a microbial system is developed for the production of high‐added‐value N‐heterocycles directly from glucose. Based on a retrosynthetic analysis, eleven genes are selected, systematically modulated, and overexpressed in three Escherichia coli strains to construct an artificial pathway to produce 5‐methyl‐2‐pyrazinecarboxylic acid, a key intermediate in the production of the important pharmaceuticals Glipizide and Acipimox. Via one‐pot tandem collaborations, the designed microbes remarkably realize high‐level production of 5‐methyl‐2‐pyrazinecarboxylic acid (6.2 ± 0.1 g L−1) and its precursor 2,5‐dimethylpyrazine (7.9 ± 0.7 g L−1). This study is the first application of cooperative microbes for the total biosynthesis of functionalized N‐heterocycles and provides new insight into integrating bioretrosynthetic principles with synthetic biology to perform complex syntheses.

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Increasing l-threonine production in Escherichia coli by engineering the glyoxylate shunt and the l-threonine biosynthesis pathway

Cited by 48 publications

References 43 publications

Expression regulation of multiple key genes to improve l-threonine in Escherichia coli

Expression regulation of multiple key genes to improve l-threonine in Escherichia coli

Metabolic Engineering of Escherichia coli

Bioretrosynthesis of Functionalized N‐Heterocycles from Glucose via One‐Pot Tandem Collaborations of Designed Microbes

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