In silico profiling of cell growth and succinate production in Escherichia coli NZN111

Jian, Xingxing; Li, Ningchuan; Zhang, Cheng Cheng; Hua, Qiang

doi:10.1186/s40643-016-0125-5

Cited by 10 publications

(8 citation statements)

References 40 publications

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“…When cultured in semi‐anaerobic conditions, EMY‐333DCBDA showed defective growth patterns compared to EMY‐33DCBDA (Figure S1a, Supporting Information). The pflB mutant strains exhibited a longer lag phase and a 33% decrease in growth, which agrees with previous research findings . Although EMY‐333DCBDA displayed poor fitness, 78% less ethanol was produced compared to EMY‐33DCBDA, and a minor amount of acetate was generated (Figure ).…”

Section: Resultssupporting

confidence: 90%

“…Furthermore, decreasing the precursor of ethanol, acetyl‐CoA, can inhibit cell growth because acetyl‐CoA is involved in fatty acid synthesis and ATP generation through the acetate formation pathway . We found that a reduced amount of ethanol was produced in the pflB mutant; however, growth defects similar to those observed in previous studies were also present …”

Section: Discussionsupporting

confidence: 74%

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Formate and Nitrate Utilization in Enterobacter aerogenes for Semi‐Anaerobic Production of Isobutanol

Jung

Kim

2017

Biotechnology Journal

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Anaerobic bioprocessing is preferred because of its economic advantages. However, low productivity and decreased growth of the host strain have limited the use of the anaerobic process. Anaerobic respiration can be applied to anoxic processing using formate and nitrate metabolism to improve the productivity of value-added metabolites. A isobutanol-producing strains is constructed using Enterobacter aerogenes as a host strain by metabolic engineering approaches. The byproduct pathway (ldhA, budA, and pflB) is knocked out, and heterologous keto-acid decarboxylase (kivD) and alcohol dehydrogenase (adhA) are expressed along with the L-valine synthesis pathway (ilvCD and budB). The pyruvate formate-lyase mutant shows decreased growth rates when cultivated in semi-anaerobic conditions, which results in a decline in productivity. When formate and nitrate are supplied in the culture medium, the growth rates and amount of isobutanol production is restored (4.4 g L , 0.23 g g glucose, 0.18 g L h ). To determine the function of the formate and nitrate coupling reaction system, the mutant strains that could not utilize formate or nitrate is contructed. Decreased growth and productivity are observed in the nitrate reductase (narG) mutant strain. This is the first report of engineering isobutanol-producing E. aerogenes to increase strain fitness via augmentation of formate and nitrate metabolism during anaerobic cultivation.

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Section: Resultssupporting

confidence: 90%

Section: Discussionsupporting

confidence: 74%

Formate and Nitrate Utilization in Enterobacter aerogenes for Semi‐Anaerobic Production of Isobutanol

Jung

Kim

2017

Biotechnology Journal

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“…Succinate producing microorganisms and fermentation strategies are developed to reduce or even finally replace petroleum-based succinate production. Natural succinate production bacteria, including Anaerobiospirillum succiniciproducens (Bretz 2015), Actinobacillus succinogenes (Carvalho et al 2016;Zou et al 2011), and Mannheimia succiniciproducens (Lee et al 2016;Ahn et al 2017), and non-native strains, such as Escherichia coli (Chen et al 2014;Meng et al 2016;Jian et al 2016;Li et al 2017b) and Corynebacterium glutamicum (Okino et al 2008;Chung et al 2017) have been studied for succinate production.…”

Section: Introductionmentioning

confidence: 99%

Production of succinate from simply purified crude glycerol by engineered Escherichia coli using two-stage fermentation

Huang

et al. 2018

Bioresour. Bioprocess.

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Background: Crude glycerol is a main by-product from biodiesel production, and efficient utilization of crude glycerol will bring significant economic and environmental benefits. However, the complex compositions of crude glycerol may impair the cellular growth and inhibit the crude glycerol consumption. Therefore, it is necessary to find a simple method to treat the crude glycerol and release the inhibition on cell metabolism. Results: The simply purified crude glycerol by activated carbon can be used as the carbon source to produce succinate in two-stage fermentation by the engineered Escherichia coli strain, MLB (ldhA − , pflB −) expressing phosphoenolpyruvate carboxykinase. In the flask experiments, succinate production from crude glycerol without treatment was less than that from pure glycerol. However, in the experiments of 1.5-L bioreactor, little succinate was produced in crude glycerol. The simply purified crude glycerol was used as carbon source for succinate production, and the glycerol consumption and succinate production were enhanced greatly. The succinate produced from the simply purified crude glycerol reached 566.0 mM, which was about ten times higher as that of non-purified one (50.3 mM). The succinate yield of the anaerobic stage achieved 0.97 mol/mol, which was 97% of the theoretical yield. Conclusion: The treatment of crude glycerol by activated carbon could effectively release the inhibition on the glycerol consumption and succinate production of the engineered E. coli strains, so that the fermentation result of the treated crude glycerol was similar as the pure glycerol. The results showed that the metabolically engineered E. coli strains have great potential to produce succinate from crude glycerol.

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“…In microbial production of many natural products, increasing precursor and cofactor availability has become an effective approach in achieving efficient production platform (Chemler, Fowler, Mchugh, & Koffas 2010;Du & Shao, 2011;Jian, Li, Zhang, & Hua, 2016;Wang, San, & Bennett, 2013). In our previous studies, efforts for increasing production of fatty acid in E. coli by breaking the TCA cycle to reduce precursor drainage and by enhancing the enzymes that involved in the fatty acid elongation cycle such as overexpression FabZ and FabA had achieved higher titers and yields (Li, Zhang, Agrawal, & San, 2012;Wu, Karanjikar, & San, 2014).…”

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confidence: 99%

Effect of NADPH availability on free fatty acid production in Escherichia coli

et al. 2017

Biotech & Bioengineering

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Microbial conversion of renewable carbon sources to free fatty acids has attracted significant attention in recent years. Accumulation of free fatty acids in Escherichia coli by overexpression of an acyl-ACP thioesterase which can break the fatty acid elongation has been well established. Various efforts have been made to increase fatty acid production in E. coli by enhancing the enzymes involved in the fatty acid synthesis cycle or host strain manipulations. The current study focused on the effect of NADPH availability on free fatty acids (FFAs) productivity. There are two reduction steps in the fatty acid elongation cycle which are catalyzed by beta keto-ACP reductase (FabG) and enoyl-ACP reductase (FabI), respectively. It is reported that FabI can use either NADH or NADPH as cofactor, while FabG only uses NADPH in E. coli. Fatty acid production dropped dramatically in the glucose-6-phosphate dehydrogenase (encoded by the zwf gene) deficient strain. Similarly, the pntB (which encodes one of the subunit of proton-translocating membrane bounded transhydrogenase PntAB) and udhA (which encodes the energy dependent cytoplasmic transhydrogenase UdhA) double mutant strain also showed an 88.8% decrease in free fatty acid production. Overexpression of PntAB and NadK restored the fatty acid production capability of these two mutant strains. These results indicated that the availability of NADPH played a very important role in fatty acid production.

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In silico profiling of cell growth and succinate production in Escherichia coli NZN111

Cited by 10 publications

References 40 publications

Formate and Nitrate Utilization in Enterobacter aerogenes for Semi‐Anaerobic Production of Isobutanol

Formate and Nitrate Utilization in Enterobacter aerogenes for Semi‐Anaerobic Production of Isobutanol

Production of succinate from simply purified crude glycerol by engineered Escherichia coli using two-stage fermentation

Effect of NADPH availability on free fatty acid production in Escherichia coli

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