Preliminary Optimization Of L-Arginine Fermentation Technology On The Genetically Engineered Bacteria Corynebacterium Glutamicum

In this paper,the reconstructed strain ATCC14067-T18-argB was used to produce L-arginine.The method of L-arginine detection was studied.The fermentation medium was studied and optimized with the Box-Bohnken method.The fed-batched cultivation with glucose was studied and the inoculation was studied.The main research contents and results are as follows:1.The High Performance Liquid Chromatography method was used to evaluate the concentration of L-arginine.The proper derivation reagent and the HPLC columns were studied and the elution program was optimized.Then test the linear relationship,the repeatability and the robust.2.Aiming at improving the production of L-arginine,several factors including soya peptone,glucose,urea,calcium carbonate and liquid volume were studied on the one-variable-at-a-time.Three main factors,soya peptone,glucose and urea were studied by the path of steepest ascent and response surface method.The optimal medium which containing soya peptone 9.0 g/L,glucose 80.0 g/L and urea 8.0 g/L was applied.The mutant ATCC14067-T18-argB could accumulate L-arginine after 72 h fermentation at 10.77 g/L,which increased 195.07%.The mutant 14067-T18-argB-E-lysE increased the yield of L-arginine 193.49%,the mutant 14067-T18-argJBD-lysE increased the yield of L-arginine 46.92 %,the mutant 14067-T18-14-21 increased the yield of L-arginine 154.53 %.3.Based on the flask-shaking batch fermentation condition,the batch fermentation was investigated with strain ATCC14067-T18-argB in 3 liter fermenter.The fed-batched cultivation with glucose was studied,the initial inoculum and the addition concentration of glucose were optimized,the yield of L-arginine increased 21.82% finally.