Study On Production Of Recombinant Human Lysozyme Via Fermentation In Pichia Pastoris

Compared of egg lysozyme usually widely utilized, human lysozyme has more safety and less byeffect to human body when it has been injected into human body. But, the row material of human lysozyme, which is only obtained from person's milk, placenta or urine, is very limitative. In order to meet the requement of market, genetically engineered yeast Pichia PastorisGS115 which boring human lysozyme gene was used to express rhLysozyme in our lab.Here we reported the results on fermentation condition optimization and purification of recombination human lysozyme.We investigated the different component of carbon and nitrogen in industry medium influence on cell growth as well as the different pH and carbon source influence on gene expression. When reduced both carbon source from 40g/L to 35g/L and nitrogen source from 10g/l to 5g/L, the cell density had not been reduced obviously compared with the traditional medium. Moreover,the inducer methanol was addited into medium at about 36hrs when the glycerol was completely exhausted. It was more favorable to express the target protein when the medium pH was maintained at pH4.0.For purification of the target protein, a couple operation of both SP-Sepharose FF strong cation-resin exchange chromatography and DEAE Sepharose FF weak anion-exchange resin chromatography had been employed to obtain the purified target protein about 50mg/L. For...