Efficient production of L-homoserine in Corynebacterium glutamicum ATCC 13032 by redistribution of metabolic flux

Li, Ning; Xu, Sha; Du, Guocheng; Chen, Jian; Zhou, Jingwen

doi:10.1016/j.bej.2020.107665

Cited by 23 publications

(25 citation statements)

References 47 publications

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“…The culture conditions were described as our previous study [ 17 ]. C. glutamicum strains were grown in LBHIS medium at 30°C.…”

Section: Methodsmentioning

confidence: 99%

“…L-Homoserine can be efficiently accumulated in C. glutamicum , indicating that OAH should also be efficiently accumulated through efficient acetyl transfer [ 19 ]. Unfortunately, the engineered C. glutamicum strain only efficiently accumulated L-homoserine but not OAH without knock-out of the metX gene in our previous studies [ 17 , 18 ], suggesting that some problems need to be solved to achieve OAH accumulation. These problems may include the total enzyme activity, specific enzyme activity, heat resistance of L-homoserine acetyltransferase (MetX), and even the supply of acetyl-CoA [ 20 – 22 ].…”

Section: Introductionmentioning

confidence: 94%

“…Reports have shown that L-homoserine and OAH have been produced efficiently in E. coli [12][13][14][15][16]. However, thus far, C. glutamicum has only achieved efficient production of L-homoserine [17,18]. L-Homoserine and acetyl-CoA are the substrates for OAH biosynthesis, whereas the production of OAH in C. glutamicum has not been reported.…”

Section: Open Accessmentioning

confidence: 99%

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O-Acetyl-L-homoserine production enhanced by pathway strengthening and acetate supplementation in Corynebacterium glutamicum

Zeng

Zhou

et al. 2022

Biotechnol Biofuels

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Background O-Acetyl-L-homoserine (OAH) is an important potential platform chemical. However, low levels of production of OAH are greatly limiting its industrial application. Furthermore, as a common and safe amino acid-producing strain, Corynebacterium glutamicum has not yet achieved efficient production of OAH. Results First, exogenous L-homoserine acetyltransferase was introduced into an L-homoserine-producing strain, resulting in the accumulation of 0.98 g/L of OAH. Second, by comparing different acetyl-CoA biosynthesis pathways and adding several feedstocks (acetate, citrate, and pantothenate), the OAH titer increased 2.3-fold to 3.2 g/L. Then, the OAH titer further increased by 62.5% when the expression of L-homoserine dehydrogenase and L-homoserine acetyltransferase was strengthened via strong promoters. Finally, the engineered strain produced 17.4 g/L of OAH in 96 h with acetate as the supplementary feedstock in a 5-L bioreactor. Conclusions This is the first report on the efficient production of OAH with C. glutamicum as the chassis, which would provide a good foundation for industrial production of OAH.

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“…The culture conditions were described as our previous study [ 17 ]. C. glutamicum strains were grown in LBHIS medium at 30°C.…”

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 94%

Section: Open Accessmentioning

confidence: 99%

See 1 more Smart Citation

O-Acetyl-L-homoserine production enhanced by pathway strengthening and acetate supplementation in Corynebacterium glutamicum

Zeng

Zhou

et al. 2022

Biotechnol Biofuels

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“…Among the BCAAs, BrnFE transports Leu and Ile more efficiently than Val (Kennerknecht et al 2002 ). It was only in later work that BrnFE was also shown to function as a Met and HomoSer exporter (Trotschel et al 2005 ; Li et al 2020 ). In the case of Met, this function was identified in a complementary approach through microarray analysis of the gene expression profile and functional testing in a Met overproducing metD -deficient C. glutamicum strain.…”

Section: Brnfe (Tc 2a7812)mentioning

confidence: 99%

“…BrnFE and its E. coli orthologues YgaZH have been overexpressed in the production of several amino acids, including Val (Park et al 2007 ), Ile (Xie et al 2012 ; Yin et al 2013 ), HomoSer (Li et al 2020 ), and Met (Park et al 2006 ; Qin et al 2015 ), in either E. coli or C. glutamicum . In these studies, addition of the exporter to strains with elevated intracellular amino acid levels resulted in increased extracellular amino acid yields by 20 to 50%.…”

Section: Brnfe (Tc 2a7812)mentioning

confidence: 99%

Microbial methionine transporters and biotechnological applications

Mohany

Totti

Naylor

et al. 2021

Appl Microbiol Biotechnol

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Methionine (Met) is an essential amino acid with commercial value in animal feed, human nutrition, and as a chemical precursor. Microbial production of Met has seen intensive investigation towards a more sustainable alternative to the chemical synthesis that currently meets the global Met demand. Indeed, efficient Met biosynthesis has been achieved in genetically modified bacteria that harbor engineered enzymes and streamlined metabolic pathways. Very recently, the export of Met as the final step during its fermentative production has been studied and optimized, primarily through identification and expression of microbial Met efflux transporters. In this mini-review, we summarize the current knowledge on four families of Met export and import transporters that have been harnessed for the production of Met and other valuable biomolecules. These families are discussed with respect to their function, gene regulation, and biotechnological applications. We cover methods for identification and characterization of Met transporters as the basis for the further engineering of these proteins and for exploration of other solute carrier families. The available arsenal of Met transporters from different species and protein families provides blueprints not only for fermentative production but also synthetic biology systems, such as molecular sensors and cell-cell communication systems. Key points • Sustainable production of methionine (Met) using microbes is actively explored. • Met transporters of four families increase production yield and specificity. • Further applications include other biosynthetic pathways and synthetic biology.

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Optimization of CRISPR‐Cas9 through promoter replacement and efficient production of L‐homoserine in Corynebacterium glutamicum

Wang

et al. 2021

Biotechnology Journal

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Background Corynebacterium glutamicum is an important chassis for industrial applications. The low efficiency of commonly used genome editing methods for C. glutamicum limits the rapid multiple engineering of the bacterium. Main Methods and Major Results In this study, chromosome‐borne expression of cas9 and recET from Escherichia coli K12‐MG1655 was achieved to avoid toxicity to the strain, increase the probability of homologous recombination, and reduce loss of viability caused by double‐strand breaks. Constitutive strong promoters, such as P45, Ptrc, and PH36, were used to replace PglyA and to expand the application of the CRISPR‐Cas9 system. By using this system, a C. glutamicum strain producing L‐homoserine to 22.1 g per L in a 5‐L bioreactor after 96 h was obtained. Conclusions and Implications Through the application of visualized fluorescent protein, the process of plasmid curing was optimized, obtain a continuous and rapid CRISPR‐Cas9 genome editing system. The method described here could be useful to construct C. glutamicum mutant rapidly.

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Efficient production of L-homoserine in Corynebacterium glutamicum ATCC 13032 by redistribution of metabolic flux

Cited by 23 publications

References 47 publications

O-Acetyl-L-homoserine production enhanced by pathway strengthening and acetate supplementation in Corynebacterium glutamicum

O-Acetyl-L-homoserine production enhanced by pathway strengthening and acetate supplementation in Corynebacterium glutamicum

Microbial methionine transporters and biotechnological applications

Optimization of CRISPR‐Cas9 through promoter replacement and efficient production of L‐homoserine in Corynebacterium glutamicum

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