食品科学 ›› 2017, Vol. 38 ›› Issue (10): 123-129.doi: 10.7506/spkx1002-6630-201710021

• 生物工程 • 上一篇    下一篇

壳聚糖-埃洛石纳米管微球的制备及其对 木瓜蛋白酶的固定化

丁利君,瞿叶辉,刘 丹   

  1. 广东工业大学轻工化工学院,广东 广州 510006
  • 出版日期:2017-05-25 发布日期:2017-05-23
  • 基金资助:
    广东省教育部产学研合作重大专项(2013A090100009)

Preparation of Chitosan-Halloysites Nanotube Microspheres and Their Application in Immobilization of Papain

DING Lijun, QU Yehui, LIU Dan   

  1. School of Chemical Engineering and Light Industry, Guangdong University of Technology, Guangzhou 510006, China
  • Online:2017-05-25 Published:2017-05-23

摘要: 运用交联-吸附法制备壳聚糖-埃洛石纳米管(chitosan-halloysites nanotube,CTS-HNTs)复合微球固定木瓜 蛋白酶,并通过傅里叶红外光谱、扫描电子显微镜、荧光标记等方法进行表征。2% CTS+1% HNTs制备的微球对 木瓜蛋白酶的固定量最高。固定化条件为木瓜蛋白酶质量浓度1 mg/mL、固定化时间10 h。固定化木瓜蛋白酶最适 pH 6.8(游离酶pH 7.2)、最适温度60 ℃(游离酶50 ℃),保存30 d该酶相对活性为62%(游离酶27%),使用4 次 后,木瓜蛋白酶的相对活性仍然保留26.26%。CTS-HNTs微球固定化木瓜蛋白酶耐贮存,操作稳定性强,可以提高 酶的利用率,降低酶解的成本,提高生产效率。

关键词: 壳聚糖-埃洛石, 微球, 固定化, 木瓜蛋白酶

Abstract: Chitosan-halloysites nanotube (CTS-HNTs) composite microspheres were successfully prepared by crosslinkingadsorption method, and used to immobilize papain. The successful immobilization of papain on CTS-HNTs composite microspheres was confirmed through infrared spectroscopy, scanning electron microscopy, and fluorescent labeling. The microspheres prepared with 2% CTS and 1% HNTs displayed the highest immobilization efficiency for papain. The optimal immobilization conditions were determined as 1 mg/mL and 10 h for papain concentration and immobilization time, respectively. The optimal pH and temperature for the immobilized enzyme were 6.8 and 60 ℃, respectively, while those for the free enzyme were 7.2 and 50 ℃, respectively. After 30 days of storage, the relative activity of the immobilized and free papain was 62% and 27%, respectively. The immobilized enzyme retained 26.26% of the initial activity after fourth repeated use. The immobilized papain had good storage stability and operational stability, enhancing the utility efficiency of the enzyme, cutting down the cost of enzymatic hydrolysis and enhancing the production efficiency.

Key words: chitosan-halloysites, composite microspheres, immobilized enzyme, papain

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