表达禽网状内皮组织增生病病毒ggpp90蛋白的毕赤酵母基因工程菌发酵工艺优化研究

Optimization of Fermentation Conditions of Recombinant Pichia pastoris Strain Expressing Reticuloendotheliosis Virus gp90 Protein

禽网状内皮组织增生病(RE)是危害养禽业的重要传染病,基因工程亚单位疫苗是防治该病的最佳选择。研究对表达REV主要保护性抗原基因gp90的重组毕赤酵母基因工程菌株进行了发酵工艺优化,以实现重组gp90蛋白的高效稳定表达。发酵培养基、诱导温度和诱导酸碱度的优化结果表明,当诱导温度为28℃,诱导pH6.0,基础盐培养基中CaSO4、MgSO4、K2SO43种硫酸盐的使用量降低至常用浓度的50%时,目的蛋白获得高水平稳定表达,表达的重组蛋白具有良好生物活性和稳定性。本系列优化试验为重组gp90蛋白制备基因工程亚单位疫苗的产业化奠定了基础。

Reticuloendotheliosis is an significant infectious disease in poultry industry. The best approach for prevention of this disease is the use of subunit vaccines. In this study,the fermentation conditions of the recombinant Pichia strain expressing the reticuloendotheliosis virus（REV）gp90 protein was optimized so as to increase the protein production.The culture medium,temperature and pH used in induction process were optimized and optimal parameters were determined.The results showed that the optimal medium was the basic salt medium（BSM）with the concentration of CaSO4,MgSO4 and K2SO4 decrease by 50% compared to the common BSM. And,gp90 protein was expressed at the highest level when the culture temperature was 28 ℃ and pH was 6.0. This study laid the foundations for the production and commercial use of recombinant subunit vaccine against REV.