Extraction of papain in aqueous two-phase polyethylene glycol/sodium sulfate systems
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摘要: 采用聚乙二醇(PEG)/硫酸钠(Na2SO4)双水相体系对木瓜蛋白酶进行分离提取。考察各成相试剂及pH对木瓜蛋白酶活性的影响,研究体系中不同相对分子量PEG、硫酸钠和PEG的质量分数、pH、酶添加量等因素对木瓜蛋白酶在两相体系中分配行为的影响,采用响应面法优化双水相萃取木瓜蛋白酶的最佳分离条件,并进行了实验室规模的放大。结果表明:Na2SO4的质量分数对分配行为的影响极显著,其次是PEG的质量分数,pH对分配行为的影响不显著。当两相体系中PEG的相对分子量为6000,质量分数为16.0%、Na2SO4质量分数为18.0%、pH7.0、酶添加量为1.0mg/g时,木瓜蛋白酶在两相体系中的分配系数最高可达3.1,可获得木瓜蛋白酶活性回收率82.6%。实验证明:响应面法优化双水相萃取木瓜蛋白酶的条件是可行的,可用于实际预测。Abstract: The papain was separated and extracted by PEG/Na2SO4 aqueous two-phase system in this study.The influence of reagent and pH value on activity of papain were investigated, in the meantime, the influence of PEG with different molecular weights, the mass fraction of Na2SO4 and PEG, pH and enzyme added on distribution behavior in aqueous two-phase system was discussed. The separation condition of papain in aqueous two-phase system was optimizated by response surface method, and amplified in laboratory scale.The study showed that:the effect of Na2SO4 mass fraction on the distribution behavior was significant, followed by PEG mass fraction, the effect of pH on the distribution behavior was not significant. The partition coefficient and recovery rate of papain in aqueous two-phase system achieved 3.1 and 82.6% under the condition of molecular weights of PEG6000, mass fraction of PEG 16.0%, mass fraction of Na2SO418.0%, pH7.0, enzyme added 1.0mg/g. The results showed that:the optimization on extraction of papain by aqueous two-phase system was feasible and could be used to predict the actual.
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