Optimization of Protein Extraction from Hepatopancreas of Litopenaeus vannamei and Effect of Environmental Stress during Waterless Transportation on Apoptosis-Related Proteins
ZHENG Xiaoxian, WU Jiaxin, XU Defeng, SUN Lijun, WANG Yaling, QIN Xiaoming, FAN Xiuping
1.Guangdong Provincial Engineering Technology Research Center of Marine Food, Guangdong Provincial Key Laboratory of Aquatic Product Processing and Safety, College of Food Science and Technology, Guangdong Ocean University, Zhanjiang 524088, China; 2.Collaborative Innovation Center of Seafood Deep Processing, Dalian Polytechnic University, Dalian 116034, China
Abstract：In the present study, in order to improve the reliability of Western blotting results, the effects of extraction buffers, grinding methods, inhibitor type and concentration, and extraction time on protein extraction from the hepatopancreas of Litopenaeus vannamei were explored by considering protein dissolution rate and the completeness of protein bands in electrophoretic gels. Next, the expression levels of apoptosis-related proteins in the hepatopancreas of Litopenaeus vannamei were detected with Western blotting. The results showed that the protein dissolution rate was higher using RIPA lysis buffer compared with phosphate buffer saline (PBS) or water as the extraction solvent. Electric homogenization and liquid nitrogen grinding provided more complete protein bands. The degradation of proteins by endogenous proteases was obviously suppressed by addition of 4% protease and phosphatase inhibitor cocktail. In addition, Western blotting results revealed that in addition to inducing Litopenaeus vannamei into dormancy, low temperatures induced slight apoptosis in the hepatopancreas tissue. Furthermore, the apoptosis level increased with waterless duration and this effect was alleviated after removal of environmental stress.
郑晓娴，吴嘉鑫，徐德峰，孙力军，王雅玲，秦小明，范秀萍. 南美白对虾肝胰腺蛋白提取方法优化及无水保活环境胁迫对细胞凋亡蛋白的影响[J]. 肉类研究, 2022, 36(6): 9-15.
ZHENG Xiaoxian, WU Jiaxin, XU Defeng, SUN Lijun, WANG Yaling, QIN Xiaoming, FAN Xiuping. Optimization of Protein Extraction from Hepatopancreas of Litopenaeus vannamei and Effect of Environmental Stress during Waterless Transportation on Apoptosis-Related Proteins. Meat Research, 2022, 36(6): 9-15.
2022, Vol. 36 
