高等学校化学学报 ›› 2019, Vol. 40 ›› Issue (9): 1888-1896.doi: 10.7503/cjcu20190185

• 有机化学 • 上一篇    下一篇

吸附-纤维素覆膜联合固定化酶

吴蓉1,2,董其惠1,2,孙伊伊1,苏二正1,2,*()   

  1. 1. 南京林业大学轻工与食品学院食品科学与工程系
    2. 南方现代林业协同创新中心, 南京 210037
  • 收稿日期:2019-03-27 出版日期:2019-09-10 发布日期:2019-07-29
  • 通讯作者: 苏二正 E-mail:ezhsu@njfu.edu.cn
  • 基金资助:
    江苏省“六大人才高峰”高层次人才项目(2015-JY-016);中国博士后科学基金(2016M600417, 2017T100373);江苏省第五期“333工程”培养资金(BRA20174586)

Efficient Enzyme Immobilization by Combining Adsorption and Cellulose Membrane Coating

WU Rong1,2,DONG Qihui1,2,SUN Yiyi1,SU Erzheng1,2,*()   

  1. 1. Department of Food Science and Technology, College of Light Industry and Food Engineering, Nanjing Forestry University
    2. Co-innovation Center for the Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing 210037, China
  • Received:2019-03-27 Online:2019-09-10 Published:2019-07-29
  • Contact: SU Erzheng E-mail:ezhsu@njfu.edu.cn
  • Supported by:
    ? Supported by the Six Talent Peaks Project in Jiangsu Province, China(2015-JY-016);the China Postdoctoral Science Foundation(2016M600417, 2017T100373);the 333 Project of Jiangsu Province, China(BRA20174586)

摘要:

以食品工业中常用的木瓜蛋白酶为模式酶, 建立了吸附-纤维素覆膜联合固定化酶方法. 通过对吸附载体类别、 纤维素种类及溶剂、 保护剂种类及其浓度、 干燥方式及时间等的优化, 得到最佳的吸附-纤维素覆膜联合固定化酶工艺. 以硅藻土或HPD-417(大孔树脂)作为吸附载体, 甲基纤维素(分子量40000~50000)丙酮溶液作为覆膜溶液, 加入6%(质量分数)的聚乙二醇或麦芽糖作为覆膜保护剂, 于4 ℃干燥9 h, 制得固定化木瓜蛋白酶, 硅藻土吸附-纤维素覆膜固定化酶酶活回收率达到96.50%, HPD-417吸附-纤维素覆膜固定化酶酶活回收率达到93.92%. 对吸附-纤维素覆膜固定化酶的性质进行了研究, 发现纤维素覆膜后固定化酶具有良好的热稳定性, 于80 ℃下保存12 h后, 固定化酶活残余率仍然能保持90%左右; 在pH=4.5~9.5的范围内, 固定化酶的稳定性较好; 连续使用9次后, 固定化酶活残余率仍能保持95%左右.

关键词: 固定化酶, 吸附, 纤维素覆膜, 联合固定化, 甲基纤维素, 保护剂, 木瓜蛋白酶

Abstract:

The papain was used as a model enzyme to develop an immobilized enzyme method based on combination of adsorption and cellulose membrane coating. The type of adsorption carries, cellulose category and the solvent type for dissolving cellulose, protectant category and concentration, drying mode and time were systematically optimized. The best adsorption-cellulose coating combined immobilization process was adopting diatomite or HPD-417(macroporous resin) as adsorption carrier, methylcellulose(molecular weight 40000—50000) acetone solution as coating solution, 6%(mass fraction) polyethylene glycol or maltose as the protectant for membrane coating, and drying at 4 ℃ for 9 h. The prepared immobilized papain showed high enzyme activity recovery rate, which attained to 96.50% for diatomite adsorbing-cellulose coating immobilized papain and 93.92% for HPD-417 adsorbing-cellulose coating immobilized papain. Finally, the properties of the immobilized enzyme were investigated. The results showed that the adsorbing-cellulose coating immobilized papain had better thermal stability. After being stored at 80 ℃ for 12 h, the residual activity of the immobilized enzyme remained about 90%. In the range of pH=4.5—9.5, the stability of the immobilized enzyme was better. After being continuously used for 9 batches, the residual activity of the immobilized enzyme remained about 95%.

Key words: Enzyme immobilization, Adsorption, Cellulose coating, Combined immobilization, Methyl cellulose, Protectant, Papain

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