Liquid-liquid extraction of bromelain and polyphenol oxidase using aqueous two-phase system
Liquid-liquid extraction of bromelain and polyphenol oxidase using aqueous two-phase system
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DOI：
10.1016/j.cep.2007.08.006
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发表时间：
2008-01-01
影响因子：
4.3
通讯作者：
Raghavarao, K. S. M. S.
中科院分区：
文献类型：
--
作者：
Babu, B. Ravindra;Rastogi, N. K.;Raghavarao, K. S. M. S.
首次采用双水相萃取法对菠萝蛋白酶和多酚氧化酶混合物进行分离纯化。研究了成相聚合物分子量（聚乙二醇1500 ~ 20000）、成相组分浓度（聚乙二醇12 ~ 18%，w/w，磷酸钾14 ~ 20%，w/w）和体系pH（6.0 ~ 9.0）等参数对纯化所需菠萝蛋白酶和多酚氧化酶差异分配的影响。菠萝蛋白酶优先分配到顶部（聚乙二醇）相，而多酚氧化酶优先分配到底部（磷酸钾）相。两种酶的分配系数随聚乙二醇分子量的增加而减小。发现体系的相组成对酶的分配和纯化程度有显著影响。聚乙二醇/磷酸钾体系（由18% PEG 1500和14%磷酸盐组成）对菠萝蛋白酶的活性回收率约为228%，纯度提高4.0倍；对多酚氧化酶的活性回收率约为90%，纯度提高2.7倍。(c) 2007爱思唯尔b.v.。版权所有。
Aqueous two-phase extraction is employed for the first time for separation and purification of mixture of enzymes (bromelain and polyphenol oxidase) from the pineapple (Ananas comosus L. Merr.). Influence of several parameters such as phase forming polyrner molecular weight (polyethylene glycol 1500-20,000), concentration of phase forming components (polyethylene glycol, 12-18%, w/w, and potassium phosphate, 14-20%, w/w) and system pH (6.0-9.0) on differential partitioning of bromelain and polyphenol oxidase required for the purification is studied. Bromelain preferentially partitioned to the top (polyethylene glycol) phase, while polyphenol oxidase to the bottom (potassium phosphate) phase. Partition coefficient of both the enzymes decreased with an increase in molecular weight of polyethylene glycol. Phase composition of the system was found to have significant effect on enzyme partitioning and degree of purification. The polyethylene glycol/potassium phosphate system (comprising of 18% PEG 1500 and 14% phosphate) resulted in about 228% activity recovery and 4.0-fold increase in purity in case of bromelain and about 90% activity recovery and 2.7-fold increase in purity of polyphenol oxidase. (c) 2007 Elsevier B.V.. All rights reserved.