Increasing<scp>l</scp>-isoleucine production in<i>Corynebacterium glutamicum</i>by overexpressing global regulator Lrp and two-component export system BrnFE

Yin, Lianghong; Shi, Feng; Hu, Xiaoqing; Chen, C.; Wang, X.

doi:10.1111/jam.12141

Cited by 50 publications

(31 citation statements)

References 35 publications

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“…In C. glutamicum ssp. lactofermentum, Ile is synthesized from L-aspartate and pyruvate by a ten-step reaction (Shi et al 2012) and then exported effectively by the transporter BrnFE (Yin et al 2013). Sufficient supply of OAA and pyruvate is important for Ile biosynthesis.…”

Section: Discussionmentioning

confidence: 99%

4-Hydroxyisoleucine production of recombinant Corynebacterium glutamicum ssp. lactofermentum under optimal corn steep liquor limitation

Shi

Niu

Fang

2015

Appl Microbiol Biotechnol

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4-Hydroxyisoleucine (4-HIL) is a nonproteinogenic amino acid that exhibits insulinotropic biological activity. Here, L-isoleucine dioxygenase gene (ido) derived from Bacillus thuringiensis YBT-1520 was cloned and expressed in an L-isoleucine-producing strain, Corynebacterium glutamicum ssp. lactofermentum SN01, in order to directly convert its endogenous L-isoleucine (Ile) into 4-HIL through single-step fermentation. The effects of corn steep liquor limitation as well as ido and truncated idoΔ6 overexpression on 4-HIL production were researched. 4-HIL production by ido-overexpressing strain was improved to 65.44 ± 2.27 mM after fermented for 144 h under corn steep liquor-subsufficient condition, obviously higher than that under corn steep liquor-rich and insufficient conditions. The conversion ratio of Ile to 4-HIL increased to 0.85 mol/mol. In addition, 4-HIL production by ido-overexpressing strain was higher than that by idoΔ6-overexpressing strain, in accord with the relatively higher affinity of Ido as compared to IdoΔ6. This research generated a novel system for 4-HIL de novo biosynthesis and demonstrated corn steep liquor limitation as a useful strategy for improving 4-HIL production in recombinant C. glutamicum ssp. lactofermentum.

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Section: Discussionmentioning

confidence: 99%

4-Hydroxyisoleucine production of recombinant Corynebacterium glutamicum ssp. lactofermentum under optimal corn steep liquor limitation

Shi

Niu

Fang

2015

Appl Microbiol Biotechnol

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“…Another expectation for improving process productivity derives from recently revealed research on the global regulators governing the expression of genes involved in sugar uptake, glycolysis, the pentose phosphate pathway, and the TCA cycle [64,68,76]. On l-isoleucine production, overexpression of the gene encoding the global regulator Lrp (which regulates the expression of several operons involved in the metabolism of branched-chain amino acid) was shown to increase the productivity [87].…”

Section: Discussionmentioning

confidence: 99%

Recent advances in the metabolic engineering of Corynebacterium glutamicum for the production of lactate and succinate from renewable resources

Tsuge

Hasunuma

Kondo

2015

Journal of Industrial Microbiology and Biotechnology

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Recent increasing attention to environmental issues and the shortage of oil resources have spurred political and industrial interest in the development of environmental friendly and cost-effective processes for the production of bio-based chemicals from renewable resources. Thus, microbial production of commercially important chemicals is viewed as a desirable way to replace current petrochemical production. Corynebacterium glutamicum, a Gram-positive soil bacterium, is one of the most important industrial microorganisms as a platform for the production of various amino acids. Recent research has explored the use of C. glutamicum as a potential cell factory for producing organic acids such as lactate and succinate, both of which are commercially important bulk chemicals. Here, we summarize current understanding in this field and recent metabolic engineering efforts to develop C. glutamicum strains that efficiently produce L- and D-lactate, and succinate from renewable resources.

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“…For example, overexpressing the feedbackresistant TD and AHAS in C. glutamicum IWJ001 produced 30.7 g/L l-isoleucine after 72 H fermentation in a 3-L fermentor [8]. Overexpressing the global regulator Lrp, the export system BrnFE, and the ribosome elongation and recycling factors could also increase L-isoleucine production in C. glutamicum [9][10][11][12].…”

Section: Metabolic Pathways Associated With the L-isoleucine Biosynthmentioning

confidence: 96%

Enhancing pentose phosphate pathway in Corynebacterium glutamicum to improve l‐isoleucine production

Wang

et al. 2016

Biotech and App Biochem

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Three genes, gnd, pgl, and fbp, relevant to the pentose phosphate pathway (PPP) were overexpressed in Corynebacterium glutamicum IWJ001, leading to increase of l-isoleucine production. The transcriptional levels of gnd, pgl, and fbp significantly increased in IWJ001/pDXW-8-gnd-fbp-pgl. Compared with the control strain IWJ001/pDXW-8, intracellular NADPH/NADP ratios in IWJ001/pDXW-8-gnd and IWJ001/pDXW-8-gnd-fbp cells grown for 36 H increased threefold and fourfold, respectively, indicating that overexpression of gnd and fbp redirected the carbon flux to PPP. Intracellular NADPH/NADP ratio in IWJ001/pDXW-8-gnd-fbp-pgl grown for 36 H was similar to IWJ001/pDXW-8, suggesting that the NADPH produced by PPP could be quickly consumed for l-isoleucine production. 10.9 and 28.96 g/L of l-isoleucine was produced in IWJ001/pDXW-8-gnd-fbp-pgl in shake flask cultivation and fed-batch fermentation, respectively. In addition, IWJ001/pDXW-8-gnd-fbp-pgl grew fast, its dry cell weight reached 49 g/L after 48 H, whereas the start strain IWJ001/pDXW-8 reached only 40 g/L. After 96 H fermentation, l-isoleucine yield on glucose in IWJ001/pDXW-8-gnd-fbp-pgl reached 0.138 g/g. The results demonstrate that carbon flux redirection to PPP is an efficient approach to enhance l-isoleucine production in C. glutamicum.

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Increasingl-isoleucine production inCorynebacterium glutamicumby overexpressing global regulator Lrp and two-component export system BrnFE

Cited by 50 publications

References 35 publications

4-Hydroxyisoleucine production of recombinant Corynebacterium glutamicum ssp. lactofermentum under optimal corn steep liquor limitation

4-Hydroxyisoleucine production of recombinant Corynebacterium glutamicum ssp. lactofermentum under optimal corn steep liquor limitation

Recent advances in the metabolic engineering of Corynebacterium glutamicum for the production of lactate and succinate from renewable resources

Enhancing pentose phosphate pathway in Corynebacterium glutamicum to improve l‐isoleucine production

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