斜带石斑鱼生长激素基因重组蛋白的发酵  

Fermentation of Recombinant Protein of Growth Hormone Gene in Orange-spotted Grouper

作　　者：吕晴霁 潘忠超 石和荣 杨慧荣[1] 王树启[3] 赵会宏[1] LU Qingji PAN Zhongchao SHI Herong YANG HuirongI WANG Shuqi ZHAO(Animal Science College, South China Agricultural University, Guangzhou 510642, China Guangdong Provincial Key Laboratory of Marine Biotechnology, Shantou 515063, China Guangdong Haid Institute of Animal Husbandry Veterinary,Guangzhou 510630.China)

机构地区：[1]华南农业大学动物科学学院,广东广州510642 [2]广东海大畜牧兽医研究院有限公司,广东广州510630 [3]汕头大学广东省海洋生物技术重点实验室,广东汕头515063

出　　处：《水产科学》2017年第1期88-93,共6页Fisheries Science

基　　金：广东省科技计划项目(2015A020208002);广东省海洋生物技术重点实验室开放基金资助项目(GPKLMB201301);广东省海洋渔业科技推广专项(A201501A03;A201501A09;A201501A10)

摘　　要：将含有石斑鱼生长激素基因的毕赤酵母X33进行震荡培养后,装入≥50L的自控罐中,28~30.5℃、pH 5.5~6.0、溶解氧≥30mg/L条件下发酵84h,生产石斑鱼生长激素基因重组蛋白。发酵期间,通过甲醇诱导毕赤酵母获得目的蛋白的高效表达,最后通过Western Blot和SDS-PAGE聚丙酰胺凝胶电泳对目的蛋白进行定性定量检测。最终获得目的蛋白表达量约为1.5~2.0g/L,约占胞外可溶性总蛋白的16.4%。本试验用自控罐发酵生产含有斜带石斑鱼生长激素基因重组蛋白的毕赤酵母生产工艺,为实现工业化生产鱼类生长激素饵料添加剂奠定基础。Yeast X33 Pichia pastoris were applied to express recombinant of growth hormone in orange- spotted grouper Epinephelus coioides cultivated in shaking flask first, and than fermented in 50 liters or bigger auto-control fermentator under conditions of temperature from 28℃ to 30.5 ℃ ,and pH 5.5-6.0 for 76 hours. During the fermentation period, the Yeast X33 were induced to express high amount protein by methanol, and the targets were detected qualitatively and quantitatively by Western Blot and SDS- PAGE. The concentration of target protein was found to be changed from 1.5 g/L to 2.0 g/L, accounting for 16.4% of total soluble protein in vitro. The fementation technology which producted recombinant or- ange-spotted grouper growth hormone in yeast X33 P. pastoris by an automatic control fermentor lays the foundation of industrial production of fish growth hormone feed additives.

关 键 词：斜带石斑鱼 生长激素基因重组蛋白 毕赤酵母 发酵 

分 类 号：S965.334[农业科学—水产养殖]