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Engineering Escherichia coli for D-allulose biosynthesis from glycerol
Qiang Guo1, Zhen-Xing Dong1, Xuan Luo1
1College of Chemical Engineering, Fujian Engineering Research Center of Advanced Manufacturing Technology for Fine Chemicals, Fuzhou University, Fuzhou 350108, China.
Journal of Biotechnology
|August 24, 2024
Summary
Microbial fermentation enables efficient D-allulose production from glycerol using engineered Escherichia coli. This novel approach achieved a high titer of 7.02 g/L, offering a promising alternative to traditional enzyme catalysis for this low-calorie sweetener.
Area of Science:
- Biotechnology
- Metabolic Engineering
- Synthetic Biology
Background:
- D-allulose is a naturally occurring monosaccharide with valuable low-calorie sweetener properties.
- Current production methods primarily rely on in vitro enzyme catalysis.
- Microbial fermentation presents an advantageous alternative by integrating enzyme production and synthesis.
Purpose of the Study:
- To develop an efficient microbial fermentation process for D-allulose biosynthesis.
- To engineer Escherichia coli for the production of D-allulose from glycerol.
- To optimize the D-allulose synthesis pathway and minimize its consumption.
Main Methods:
- Metabolic engineering of Escherichia coli by introducing D-allulose synthesis genes (FbaA, Fbp, AlsE, A6PP).
- Disruption of key metabolic pathways (PfkA, PfkB, Pgi) to prevent intermediate (fructose-6-phosphate) loss.
- Inactivation of D-allulose consuming enzymes (GalE, FryA).
- Implementation of a fed-batch fermentation strategy.
Main Results:
- Successful construction of a D-allulose synthesis pathway in engineered E. coli.
- Achieved a D-allulose titer of 7.02 g/L.
- Obtained a maximum D-allulose yield of 0.287 g/g from glycerol.
- Demonstrated reduced D-allulose consumption by the engineered strain.
Conclusions:
- Metabolically engineered Escherichia coli can efficiently produce D-allulose from glycerol via microbial fermentation.
- The developed fed-batch process significantly enhances D-allulose production.
- This engineered microbial cell factory offers a scalable and sustainable method for D-allulose synthesis.
