生物加工过程2016，Vol.14Issue(4)：17-23,7.DOI:10.3969/j.issn.1672-3678.2016.04.004
聚乙二醇共沉淀物复溶萃取辣根过氧化物酶的条件优化
Optimization of extraction of horseradish peroxidase by aqueous two-phase system with polyethylene glycol co-precipitation and resolution
摘要
Abstract
To improve the extraction and purity of horseradish peroxidase ( HRP ) by aqueous two⁃phase extraction, polyethylene glycol( PEG6000) was used to precipitate the protein enriched with HRP in the high saturation ammonium sulfate. Then, an aqueous two phase system of PEG/phosphate was formed based on the Co⁃precipitation resolution to extracted higher composition HRP . The influence of different pH,PEG concentration,( NH4 ) 2 SO4 saturation on the enzyme activity and the effect of different liquid⁃solid ratio, NaCl and pH concentration on the distribution of the target enzyme in aqueous two⁃phase system were explored by single factor experiments. The extraction conditions were optimized by response surface method. The optimal extraction conditions were:liquid⁃solid ratio 0. 3 mL/g, pH of phosphate buffer saline 7. 02,NaCl 42 g/L.Under the optimal extraction conditions,the recovery rate of the enzyme activity was 88. 1%,and the purity was increased by 21. 7 times. The establishment of this method is of great significance for improving the economic benefit of the biological downstream process.关键词
双水相萃取/辣根过氧化物酶/响应面法Key words
aqueous two-phase extraction/horseradish peroxidase/response surface method分类
化学工程引用本文复制引用
王作铭,李英东,高敏强,赵雪君,和文燕,明子,王文佳,陈军..聚乙二醇共沉淀物复溶萃取辣根过氧化物酶的条件优化[J].生物加工过程,2016,14(4):17-23,7.基金项目
上海市大学生创新活动计划项目 （）
