CIESC Journal ›› 2015, Vol. 66 ›› Issue (1): 179-185.DOI: 10.11949/j.issn.0438-1157.20140555

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Partitioning behavior and structural characterization of papain in ionic liquid aqueous two-phase system

WANG Weitao, JIANG Zhiguo, ZHANG Haide, PENG Jian, XU Yinghao, DONG Anhua, YANG Xuefang, JIANG Xinxin   

  1. School of Food Science and Technology, Hainan University, Haikou 570228, Hainan, China
  • Received:2014-04-08 Revised:2014-10-12 Online:2015-01-05 Published:2015-01-05
  • Supported by:

    supported by the National Natural Science Foundation of China (31260401).

木瓜蛋白酶在离子液体双水相中的分配行为

王伟涛, 蒋志国, 张海德, 彭健, 许英豪, 董安华, 杨雪芳, 蒋欣欣   

  1. 海南大学食品学院, 海南 海口 570228
  • 通讯作者: 张海德
  • 基金资助:

    国家自然科学基金项目(31260401)。

Abstract:

Papain was extracted by ionic liquid aqueous two-phase system. Firstly, the influence of concentration, pH, and temperature of ionic liquid on the activity of papain was investigated. Secondly, the effects of different ionic liquid aqueous two-phase systems, alkyl chain lengths and concentrations of the ionic liquid, dosage of papain, pH, temperature on the partitioning behavior of papain were discussed. [C4mim]Cl and [C4mim]Br systems were better than [C4mim]BF4 system in extraction of papain, and it was disadvantageous to extract papain at a high temperature (60℃ or higher). Activity recovery of enzyme reached 95.16% and purification factor reached 1.5 under the optimum conditions: [C4mim]Cl 0.25 g·ml-1, K2HPO4 0.35 g·ml-1, pH 8.0, enzyme addition 2.0 mg·ml-1, 30℃. The result laid the experimental basis for further scale-up research and commercial production.

Key words: ionic liquid, aqueous two-phase system, papain, activity recovery of enzyme, purification factor, extraction, protein

摘要:

采用离子液体双水相提取木瓜蛋白酶。首先考察不同浓度、pH、温度的离子液体对木瓜蛋白酶活性的影响, 其次考察不同离子液体双水相体系、离子液体侧烷基链长度及浓度、酶添加量、pH、温度对木瓜蛋白酶分配行为的影响。结果表明:[C4mim]Cl和[C4mim]Br体系萃取木瓜蛋白酶的效果比[C4mim]BF4体系好;高温(≥60℃)对离子液体双水相体系萃取木瓜蛋白酶不利。离子液体双水相萃取木瓜蛋白酶的最佳工艺条件:0.25 g·ml-1的[C4mim]Cl, 0.35 g·ml-1的K2HPO4, pH 8.0, 酶添加量2.0 mg·ml-1, 30℃。此条件下木瓜蛋白酶的酶活性回收率达到95.16%, 纯化因子达到1.5。为今后进一步研究该体系的放大实验或规模化生产奠定了基础。

关键词: 离子液体, 双水相, 木瓜蛋白酶, 酶活性回收率, 纯化因子, 萃取, 蛋白质

CLC Number: