Abstract
Corynebacterium glutamicum has a long and successful history in the biotechnological production of l-lysine. Besides the adjustment of metabolic pathways, intracellular and extracellular transport systems are critical for the cellular metabolism of l-lysine or its by-products. Here, three amino acid transmembrane transporters, namely, GluE, BrnE/BrnF, and LysP, which are widely present in C. glutamicum strains, were each investigated by gene knockout. In comparison with that in the wild-type strain, the yield of l-lysine increased by 9.0%, 12.3%, and 10.0% after the deletion of the gluE, brnE/brnF, and lysP genes, respectively, in C. glutamicum 23,604. Moreover, the amount of by-product amino acids decreased significantly when the gluE and brnE/brnF genes were deleted. It was also demonstrated that there was no effect on the growth of the strain when the gluE or lysP gene was deleted, whereas the biomass of C. glutamicum WL1702 (ΔbrnE/ΔbrnF) in the fermentation medium was significantly reduced in comparison with that of the wild type. These results also provide useful information for enhancing the production of l-lysine or other amino acids by C. glutamicum.
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taken from the fermentation medium every 4 h to measure the l-glutamate, l-lysine, glucose and OD600 production of C. glutamicum 23604 and C. glutamicumWL1701, symbols: filled square strain C. glutamicum 23604, hollow triangle strain C. glutamicum WL1701. The standard errors are shown as bars
taken from the fermentation medium every 4 h to measure the l-glutamate concentration of C. glutamicum 23604 and C. glutamicum WL1702, symbols: filled square strain C. glutamicum 23604, filled triangle strain C. glutamicum WL1702. The standard errors are shown as bars
taken from the fermentation medium every 4 h to measure the l-lysine, glucose and OD600 of C. glutamicum 23604 and C. glutamicum WL1702, symbols: filled square strain C. glutamicum 23604, filled triangle strain C. glutamicum WL1702. The standard errors are shown as bars
taken from the fermentation medium every 4 h to measure the l-lysine concentration of C. glutamicum 23604 and C. glutamicum WL1702, symbols: filled square strain C. glutamicum 23604, filled triangle strain C. glutamicum WL1702. The standard errors are shown as bars
taken from the fermentation medium every 4 h to measure the l-lysine, glucose and OD600 production of C. glutamicum 23604 and C. glutamicum WL1703, symbols: hollow square strain C. glutamicum 23604, filled triangle strain C. glutamicum WL1703. The standard errors are shown as bars
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Abbreviations
- AcCoA:
-
Acetyl coenzyme A
- Ala:
-
l-Alanine
- Asn:
-
l-Asparagine
- Asp:
-
l-Aspartate
- AspSa:
-
Aspartate semialdehyde
- Chor:
-
Chorismate
- Cit:
-
Citrate
- Fum:
-
Fumaric acid
- G6P:
-
Glucose 6-phosphate
- GDL:
-
Gluconolactone
- Glc:
-
Glucose
- Gln:
-
l-Glutamine
- Glu:
-
l-Glutamate
- Gluc:
-
d-Gluconic acid
- Gluc6P:
-
6-Phospho-d-gluconate
- Glx:
-
Glyoxylate
- Gly:
-
l-Glycine
- Hcys:
-
Homocysteine
- His:
-
l-Histidine
- Hser:
-
Homoserine
- ICit:
-
Isocitrate
- Ile:
-
l-Isoleucine
- αKG:
-
α-Ketoglutarate
- Kval:
-
2-Ketovaline
- Leu:
-
l-Leucine
- Lys:
-
l-Lysine
- Mal:
-
Malic acid
- OA:
-
Oxaloacetic acid
- Met:
-
l-Methionine
- Orn:
-
Ornithine
- PEP:
-
Phosphoenolpyruvate
- PG3:
-
Glycerate 3-phosphate
- Phe:
-
l-Phenylalanine
- Pro:
-
l-Proline
- PRPP:
-
5-Phospho-d-ribosylpyrophosphate
- Pyr:
-
Pyruvate
- R5P:
-
Ribulose 5-phosphate
- Ser:
-
l-Serine
- Suc:
-
Succinate
- SucCoA:
-
Succinate coenzyme A
- Thr:
-
l-Threonine
- Trp:
-
l-Tryptophan
- Tyr:
-
l-Tyrosine
- Val:
-
l-Valine
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Acknowledgements
This work was supported by the National Science Foundation of China (31801527), Focus on Research and Development Plan in Shandong Province (2019JZZY011003, 2018YFJH0401, 2016CYJS07A01), Taishan industry leading talent (tscy20180103), Shandong Provincial Natural Science Foundation (ZR2016CB04), Major Program of National Natural Science Foundation of Shandong (ZR2017ZB0208) and Cultivation Project of Shandong Synthetic Biotechnology Innovation Center (sdsynbio-2018-PY-02).
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Xiao, J., Wang, D., Wang, L. et al. Increasing l-lysine production in Corynebacterium glutamicum by engineering amino acid transporters. Amino Acids 52, 1363–1374 (2020). https://doi.org/10.1007/s00726-020-02893-6
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DOI: https://doi.org/10.1007/s00726-020-02893-6