乙型脑炎病毒NS1_(△63)蛋白发酵工艺优化研究  

Optimization of the Fermentation Process for the Production of JEV NS1_(△63)Protein

作　　者：陶飞飞 卞佳宁 余文捡 刘仕国 熊海月 钟鸣 孔令保[1] 王婷[1] TAO Feifei;BIAN Jianing;YU Wenjian;LIU Shiguo;XIONG Haiyue;ZHONG Ming;KONG Lingbao;WANG Ting(School of Bioscience and Bioengineering/Nanchang City Key Laboratory of Animal Virus and Genetic Engineering,Jiangxi Agricultural University,Nanchang 330045,China)

机构地区：[1]江西农业大学生物科学与工程学院/南昌市动物病毒与基因工程重点试验室,江西南昌330045

出　　处：《生物灾害科学》2023年第2期242-248,共7页Biological Disaster Science

基　　金：国家自然科学基金项目(31960699);江西省自然科学基金重点项目(20224ACB205004)。

摘　　要：【目的】乙型脑炎病毒NS1蛋白能诱导机体产生保护性免疫,具有开发成亚单位疫苗的潜力。为提高重组蛋白NS1_(△63)的原核表达量,研究进行发酵条件优化以开发稳定高产的蛋白生产工艺。【方法】采用单因素与Plackett-Burman试验对培养基成分进行优化,同时探究诱导条件对发酵的影响,并在5 L发酵罐中进行初步的工艺放大验证。【结果】最佳的培养基成分为乳糖10 g/L、酵母粉19 g/L、蛋白胨25 g/L、磷酸盐37.5 mmol/L、氯化钾1.25 g/L。最佳发酵条件是诱导温度36℃、诱导时间12 h、0.6 mmol/L IPTG、5%接种量。蛋白产量提高到优化前的3.25倍,5 L发酵罐蛋白产量可达3022.23 mg/L。【结论】研究显著提高了NS1△63蛋白的原核产量,为该蛋白应用于亚单位疫苗和检测试剂盒的相关研究奠定了基础。[Objective]The NS1 protein of Japanese encephalitis virus can induce protective immunity and has the potential to be prepared into subunit vaccines.To improve the production of NS1 protein expressed by prokaryocyte,this study intends to optimize fermentation conditions to develop the stable and high-yielding protein production process.[Method]The single factor test and Plackett-Burman test were used to optimize the medium components.Meanwhile,the effects of fermentation conditions were explored,and the preliminary test was verified in 5 L fermentor.[Result]The optimal medium compositions were determined as lactose 10 g/L,yeast powder 19 g/L,peptone 25 g/L,phosphate 37.5 mmol/L,and KCl 1.25 g/L.The optimal fermentation conditions were as follows:an induction temperature of 36℃,IPTG with 0.6 mmol/L concentration,a 5%inoculum and an induction time of 12 h.The yield of the protein was increased to 3.25-fold of the initial conditions.The protein yield reached 3022.23 mg/L in 5 L fermentor.[Conclusion]The production of the NS1_(△63)protein expressed by prokaryocyte had been significantly improved,laying a foundation for the application of this protein in related studies of subunit vaccines and detection kits.

关 键 词：乙型脑炎病毒 NS1_(△63) 大肠杆菌 发酵优化 Plackett-Burman试验 

分 类 号：S855.3[农业科学—临床兽医学] TQ920.6[农业科学—兽医学]