近平滑假丝酵母ATCC 7330中羰基还原酶双水相萃取工艺研究

Aqueous Two-Phase Extraction Process of Carbonyl Reductase in Candida parapsilosis ATCC 7330

研究了聚乙二醇(PEG)/硫酸铵[(NH4)2SO4]双水相体系萃取近平滑假丝酵母ATCC 7330粗酶液中羰基还原酶的工艺,考察了无机盐种类、PEG分子量及其质量分数、(NH4)2SO4质量分数、萃取温度、萃取时间等因素对羰基还原酶纯化倍数的影响。确定最优的双水相萃取条件为:(NH4)2SO4质量分数15%、PEG1000质量分数19%、萃取温度16℃、萃取时间15 min,在此条件下,羰基还原酶的纯化倍数可达6倍。为近平滑假丝酵母ATCC 7330中羰基还原酶在手性化合物的绿色合成中的应用奠定了基础。

We studied the extraction process of carbonyl reductase in crude enzyme of Candida parapsilosis ATCC 7330 by polyethylene glycol(PEG)/ammonium sulfate[(NH4)2SO4]aqueous two-phase system,and investigated the effects of types of inorganic salts,molecular weight and mass fraction of PEG,(NH4)2SO4 mass fraction,extraction temperature,and extraction time on the purification multiple of carbonyl reductase.The optimum aqueous two-phase extraction conditions are determined as follows:the(NH4)2SO4 mass fraction of 15%,the PEG1000 mass fraction of 19%,the extraction temperature of 16℃,and the extraction time of 15 min.Under above conditions,the purification multiple of carbonyl reductase can reach 6 times.The study lays a foundation for the application of carbonyl reductase in Candida parapsilosis ATCC 7330 in the green synthesis of chiral compounds.