酸土脂环酸芽孢杆菌可溶性全细胞蛋白提取工艺优化

研究了超声波破碎法、SDS沸水浴、溶菌酶和细胞裂解液处理对酸土脂环酸芽孢杆菌破碎和可溶性全细胞蛋白提取效果的影响,并采用SDS-PAGE电泳进行蛋白成分分析,通过单因素试验和响应面法对可溶性全细胞蛋白的提取工艺条件进行了分析与优化。结果表明:细胞裂解液处理提取效果最好;细胞裂解液处理参数对全细胞蛋白提取量有显著影响,因素影响主次顺序为液料比、处理时间、处理温度;细胞裂解液处理优化工艺参数为:处理时间37.4 min、处理温度18.9℃和液料比27.0 mL/g,可溶性全细胞蛋白提取量达到161.18 mg/g。试验所得可溶性全细胞蛋白提取回归模型高度显著(R2=0.989 2),拟合性好,可用于预测。

Optimization of Soluble Whole-cell Protein Extraction from Alicylobacillus acidoterrestris

The yield of soluble whole-cell protein from Alicylobacillus acidoterrestris DSM 3922T cells is dependent on the effectiveness of cell lysis post fermentation.Various cell lysis approaches were investigated,including ultrasound,SDS,boiling,lysozyme,and cell lysis reagent treatment,and the different samples of soluble protein were determined by SDS protein polyacrylamide gel electrophoresis.The soluble whole-cell protein by one-factor-at-a-time experiments and response surface method was optimized.The results showed that the cell lysis reagent treatment was the most effective in extracting soluble whole-cell protein.The most significant processing parameters,namely incubation temperature,reaction time,and the ratio of cell lysis reagent volume and bacteria weight(A/B ratio),were optimized by using the Box-Behnken with a quadratic regression model built by response surface methodology(RSM).The influence order of factors to protein extraction rate was: A/B ratio,time and temperature.The highest soluble whole-cell protein yield at 161.18 mg/g was attained under optimal time,temperature,and A/B ratio at 37.4 min,18.9℃,and 27.0 mL/g,respectively.The regression model of soluble whole-cell protein extraction was highly significant(R2=0.989 2),the experimental and simulated results were found to be in good agreement,which could be used to forecast.