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核农学报  2014 

响应面法优化米曲霉蛋白酶的双水相萃取条件

DOI: 10.11869/j.issn.100-8551.2014.10.1860, PP. 1860-1866

Keywords: 双水相,萃取,米曲霉,蛋白酶,响应面

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Abstract:

为获得一种快速纯化米曲霉(Aspergillusoryzae,A.oryzae)蛋白酶的方法,采用聚乙二醇(polyethyleneglycol,PEG)/盐双水相系统,从米曲霉固态发酵粗提液中萃取蛋白酶,考察PEG分子质量、盐、PEG浓度、(NH4)2SO4浓度、pH值、粗酶液加入量对蛋白酶萃取的影响,并以Box-Behnken试验设计结合响应面分析法优化了萃取条件.结果表明,选用PEG/(NH4)2SO4双水相系统的最佳提取条件为:PEG-600浓度25%,(NH4)2SO4浓度24.82%,pH值6.97,粗酶液加入量为24.0%,在此条件下蛋白酶的提取率94.74%.双水相法为纯化米曲霉A.oryzae蛋白酶提供了一种有效的方法.

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