Efficient Escherichia coli platform for L-proline biosynthesis via systematic metabolic engineering.
5 affiliations
- 1National and Local United Engineering Research Center of Industrial Microbiology and Fermentation Technology, Engineering Research Center of Industrial Microbiology of Ministry of Education, College of Life Sciences, Fujian Normal University, Fujian, People's Republic of China.
- 2National and Local United Engineering Research Center of Industrial Microbiology and Fermentation Technology, Engineering Research Center of Industrial Microbiology of Ministry of Education, College of Life Sciences, Fujian Normal University, Fujian, People's Republic of China. [email protected].
- 3State Key Laboratory of Microbial Diversity and Innovative Utilization, Institute of Microbiology, Chinese Academy of Sciences, Beijing, People's Republic of China.
- 4National and Local United Engineering Research Center of Industrial Microbiology and Fermentation Technology, Engineering Research Center of Industrial Microbiology of Ministry of Education, College of Life Sciences, Fujian Normal University, Fujian, People's Republic of China. [email protected].
- 5National and Local United Engineering Research Center of Industrial Microbiology and Fermentation Technology, Engineering Research Center of Industrial Microbiology of Ministry of Education, College of Life Sciences, Fujian Normal University, Fujian, People's Republic of China. [email protected].
Abstract
Despite its prominent industrial use for L-proline production, microbial fermentation by Corynebacterium glutamicum achieves a suboptimal yield of glucose conversion, far below the theoretical maximum of 0.63 g/g. This study aimed to develop an efficient engineered Escherichia coli chassis to produce L-proline from glucose by addressing the rate-limiting enzymatic step and high glucose conversion. The γ-glutamyl kinase encoding gene (proB) was screened by directed evolution, and the catalytic efficiency of the ScProB** variant toward L-glutamate increased 2.75-fold. After channeling 2-oxoglutarate flux and supplying sufficient NADPH, L-proline production reached 86.27 ± 2.58 g/L, with a 0.33 ± 0.01 g/g glucose yield in 10-L fed-batch fermentation. Further increasing the flux through central carbon metabolism by implementing a non-PTS glucose uptake system, amplifying the anaplerotic reaction, and redirecting the flux away from the glyoxylate cycle, the resulting strain BP23 achieved 119.89 ± 5.66 g/L L-proline, with a productivity of 1.93 ± 0.03 g/L/h. The glucose conversion yield improved to 0.49 ± 0.01 g/g after 62 h of fed-batch fermentation, indicating that the E. coli strain is a promising workhorse for producing L-proline with high glucose conversion.
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BibTeX
@article{cui2026efficient,
title = {{Efficient Escherichia coli platform for L-proline biosynthesis via systematic metabolic engineering.}},
author = {Shumei Cui and Xiaolu Hu and Xinwei Yang and Yongtao Chen and Tianqi Bao and Bingyan Wang and Yong Tao and Jianzhong Huang and Chongrong Ke},
journal = {Microbial cell factories},
year = {2026},
month = jul,
doi = {10.1186/s12934-026-03075-x},
url = {https://europepmc.org/article/MED/42958551},
note = {CC BY-NC-ND},
}
RIS
TY - JOUR
TI - Efficient Escherichia coli platform for L-proline biosynthesis via systematic metabolic engineering.
AU - Shumei Cui
AU - Xiaolu Hu
AU - Xinwei Yang
AU - Yongtao Chen
AU - Tianqi Bao
AU - Bingyan Wang
AU - Yong Tao
AU - Jianzhong Huang
AU - Chongrong Ke
PY - 2026
DA - 2026/07/29
JO - Microbial cell factories
DO - 10.1186/s12934-026-03075-x
UR - https://europepmc.org/article/MED/42958551
AB - Despite its prominent industrial use for L-proline production, microbial fermentation by Corynebacterium glutamicum achieves a suboptimal yield of glucose conversion, far below the theoretical maximum of 0.63 g/g. This study aimed to develop an efficient engineered Escherichia coli chassis to produce L-proline from glucose by addressing the rate-limiting enzymatic step and high glucose conversion. The γ-glutamyl kinase encoding gene (proB) was screened by directed evolution, and the catalytic efficiency of the ScProB** variant toward L-glutamate increased 2.75-fold. After channeling 2-oxoglutarate flux and supplying sufficient NADPH, L-proline production reached 86.27 ± 2.58 g/L, with a 0.33 ± 0.01 g/g glucose yield in 10-L fed-batch fermentation. Further increasing the flux through central carbon metabolism by implementing a non-PTS glucose uptake system, amplifying the anaplerotic reaction, and redirecting the flux away from the glyoxylate cycle, the resulting strain BP23 achieved 119.89 ± 5.66 g/L L-proline, with a productivity of 1.93 ± 0.03 g/L/h. The glucose conversion yield improved to 0.49 ± 0.01 g/g after 62 h of fed-batch fermentation, indicating that the E. coli strain is a promising workhorse for producing L-proline with high glucose conversion.
SN - 1475-2859
ER -