摘要
以新鲜生姜为原料,以生姜蛋白酶酶活为考察指标,对生姜蛋白酶的提取及纯化工艺进行研究。结果表明:以pH6.0的磷酸氢二钠-磷酸二氢钠缓冲液为提取溶剂所得酶活力较高,最佳工艺为:以pH6.0磷酸氢二钠-磷酸二氢钠缓冲液25℃提取2次,料液比1∶1;应用硫酸铵盐析及透析袋透析对粗酶液进行纯化,粗酶液再以60%硫酸铵盐析24h,透析袋(14000)低温透析12h,冻干;酶学性质研究表明:生姜蛋白酶以酪蛋白为底物其最适pH值为6.0,30℃保温30min,酶活稳定。
In this paper fresh ginger was used as raw material to investigate the extraction and purification processes of the ginger proteinase. The proteinase activity determination was coomassie brilliant blue G-250. Firstly, different extract solvents were studied, and the results showed that the ginger proteinase activity was the highest when it was pH 6.0 with sodium hydrogen phosphate-sodium and sodium dihydrogen phosphate as extract solvents. Orthogonal experiment was used to optimize the extraction process, and the results was shown that at 25%, extract 2 times with material： solvent 1：1. Finally using ammonium sulfate as salting-out salt and different degree of saturation was observed. The results were shown that when the degree of saturation was 60 % salting-out 24 h, low temperature dialysis 12 h, vacuum freeze-drying the ginger proteinase activity was the highest. The ginger proteinase characteristic was also preliminary studied, and the results indicated that the ginger proteinase, take the casein as substrate and its most suitable pH is 6.0, in 30 ℃ keeps warm for 30 min, the enzyme vigor cannot be reduced, along with temperature elevating, the enzyme activity droped.
出处
《中国酿造》
CAS
北大核心
2010年第4期98-100,共3页
China Brewing
关键词
生姜蛋白酶
提取
正交试验
盐析
透析
ginger protease
extraction
orthogonal
salting-out
dialysis
