Optimization of glucose feeding approaches for enhanced glucosamine and N-acetylglucosamine production by an engineered Escherichia coli. 2012

Xin Chen, and Long Liu, and Jianghua Li, and Jie Liu, and Guocheng Du, and Jian Chen
Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China.

In this work, a recombinant Escherichia coli was constructed by overexpressing glucosamine (GlcN) synthase and GlcN-6-P N-acetyltransferase for highly efficient production of GlcN and N-acetylglucosamine (GlcNAc). For further enhancement of GlcN and GlcNAc production, the effects of different glucose feeding strategies including constant-rate feeding, interval feeding, and exponential feeding on GlcN and GlcNAc production were investigated. The results indicated that exponential feeding resulted in relatively high cell growth rate and low acetate formation rate, while constant feeding contributed to the highest specific GlcN and GlcNAc production rate. Based on this, a multistage glucose supply approach was proposed to enhance GlcN and GlcNAc production. In the first stage (0-2 h), batch culture with initial glucose concentration of 27 g/l was conducted, whereas the second culture stage (2-10 h) was performed with exponential feeding at μ (set) = 0.20 h⁻¹, followed by feeding concentrated glucose (300 g/l) at constant rate of 32 ml/h in the third stage (10-16 h). With this time-variant glucose feeding strategy, the total GlcN and GlcNAc yield reached 69.66 g/l, which was enhanced by 1.59-fold in comparison with that of batch culture with the same total glucose concentration. The time-dependent glucose feeding approach developed here may be useful for production of other fine chemicals by recombinant E. coli.

UI MeSH Term Description Entries
D004926 Escherichia coli A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc. Alkalescens-Dispar Group,Bacillus coli,Bacterium coli,Bacterium coli commune,Diffusely Adherent Escherichia coli,E coli,EAggEC,Enteroaggregative Escherichia coli,Enterococcus coli,Diffusely Adherent E. coli,Enteroaggregative E. coli,Enteroinvasive E. coli,Enteroinvasive Escherichia coli
D005944 Glucosamine 2-Amino-2-Deoxyglucose,Dona,Dona S,Glucosamine Sulfate,Hespercorbin,Xicil,2 Amino 2 Deoxyglucose,Sulfate, Glucosamine
D005947 Glucose A primary source of energy for living organisms. It is naturally occurring and is found in fruits and other parts of plants in its free state. It is used therapeutically in fluid and nutrient replacement. Dextrose,Anhydrous Dextrose,D-Glucose,Glucose Monohydrate,Glucose, (DL)-Isomer,Glucose, (alpha-D)-Isomer,Glucose, (beta-D)-Isomer,D Glucose,Dextrose, Anhydrous,Monohydrate, Glucose
D000117 Acetylglucosamine The N-acetyl derivative of glucosamine. Acetyl Glucosamine,N-Acetyl Glucosamine,N-Acetyl-beta-D-Glucosamine,N-Acetylglucosamine,beta-N-Acetylglucosamine,2-Acetamido-2-Deoxy-D-Glucose,2-Acetamido-2-Deoxyglucose,N-Acetyl-D-Glucosamine,2 Acetamido 2 Deoxy D Glucose,2 Acetamido 2 Deoxyglucose,Glucosamine, Acetyl,Glucosamine, N-Acetyl,N Acetyl D Glucosamine,N Acetyl Glucosamine,N Acetyl beta D Glucosamine,N Acetylglucosamine,beta N Acetylglucosamine
D001709 Biotechnology Body of knowledge related to the use of organisms, cells or cell-derived constituents for the purpose of developing products which are technically, scientifically and clinically useful. Alteration of biologic function at the molecular level (i.e., GENETIC ENGINEERING) is a central focus; laboratory methods used include TRANSFECTION and CLONING technologies, sequence and structure analysis algorithms, computer databases, and gene and protein structure function analysis and prediction. Biotechnologies
D014537 Uridine Diphosphate N-Acetylglucosamine Serves as the biological precursor of insect chitin, of muramic acid in bacterial cell walls, and of sialic acids in mammalian glycoproteins. UDP Acetylglucosamine,UDPGNAc,Uridine Diphospho-N-Acetylglucosamine,Uridine Pyrophosphoacetylglucosamine,Uridine Diphosphate N Acetylglucosamine,Acetylglucosamine, UDP,Diphosphate N-Acetylglucosamine, Uridine,Diphospho-N-Acetylglucosamine, Uridine,N-Acetylglucosamine, Uridine Diphosphate,Pyrophosphoacetylglucosamine, Uridine,Uridine Diphospho N Acetylglucosamine
D015309 Glucose Clamp Technique Maintenance of a constant blood glucose level by perfusion or infusion with glucose or insulin. It is used for the study of metabolic rates (e.g., in glucose, lipid, amino acid metabolism) at constant glucose concentration. Euglycemic Clamping,Glucose Clamping,Euglycaemic Clamp,Euglycaemic Clamping,Euglycemic Clamp,Glucose Clamp,Glucose Clamp Technic,Clamp, Euglycaemic,Clamp, Euglycemic,Clamp, Glucose,Clamping, Euglycaemic,Clamping, Euglycemic,Clamping, Glucose,Clamps, Euglycaemic,Clamps, Euglycemic,Clamps, Glucose,Euglycaemic Clamps,Euglycemic Clamps,Glucose Clamp Technics,Glucose Clamp Techniques,Glucose Clamps,Technic, Glucose Clamp,Technics, Glucose Clamp,Technique, Glucose Clamp,Techniques, Glucose Clamp
D061249 Batch Cell Culture Techniques Methods for cultivation of cells, usually on a large-scale, in a closed system for the purpose of producing cells or cellular products to harvest. Batch Culture Techniques,Batch Culture,Continuous Batch Culture,Fed Batch Culture Techniques,Fed-Batch Culture,Fed-Batch Culture Techniques,Shake-Flask Culture,Batch Culture Technique,Batch Culture, Continuous,Batch Cultures,Batch Cultures, Continuous,Continuous Batch Cultures,Culture Technique, Batch,Culture Technique, Fed-Batch,Culture Techniques, Batch,Culture Techniques, Fed-Batch,Culture, Batch,Culture, Continuous Batch,Culture, Fed-Batch,Culture, Shake-Flask,Cultures, Batch,Cultures, Continuous Batch,Cultures, Fed-Batch,Cultures, Shake-Flask,Fed Batch Culture,Fed-Batch Culture Technique,Fed-Batch Cultures,Shake Flask Culture,Shake-Flask Cultures,Technique, Batch Culture,Technique, Fed-Batch Culture,Techniques, Batch Culture,Techniques, Fed-Batch Culture
D018014 Gene Transfer Techniques The introduction of functional (usually cloned) GENES into cells. A variety of techniques and naturally occurring processes are used for the gene transfer such as cell hybridization, LIPOSOMES or microcell-mediated gene transfer, ELECTROPORATION, chromosome-mediated gene transfer, TRANSFECTION, and GENETIC TRANSDUCTION. Gene transfer may result in genetically transformed cells and individual organisms. Gene Delivery Systems,Gene Transfer Technique,Transgenesis,Delivery System, Gene,Delivery Systems, Gene,Gene Delivery System,Technique, Gene Transfer,Techniques, Gene Transfer,Transfer Technique, Gene,Transfer Techniques, Gene
D030781 Organisms, Genetically Modified Organisms whose GENOME has been changed by a GENETIC ENGINEERING technique. Genetically Modified Organisms,Organisms, Transgenic,Transgenic Organisms,GMO Organism,GMO Organisms,Genetically Engineered Organisms,Genetically Modified Organism,Modified Organism, Genetically,Organism, Genetically Modified,Organism, Transgenic,Organisms, Genetically Engineered

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