食品科学

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交联壳聚糖载体固定化柚苷酶工艺

雷生姣,余海立,黄 超,郑兴龙   

  1. 三峡大学化学与生命科学学院,湖北 宜昌 443002
  • 出版日期:2013-12-15 发布日期:2014-01-03
  • 通讯作者: 雷生姣
  • 基金资助:

    宜昌市夷陵区科学技术局合作研究项目(0720123200);三峡大学博士科研启动基金项目(0620120050)

Cross-linked Chitosan as a Carrier for Immobilization of Naringinase

LEI Sheng-jiao,YU Hai-li,HUANG Chao,ZHENG Xing-long   

  1. College of Chemistry and Life Science, China Three Gorges University, Yichang 443002, China
  • Online:2013-12-15 Published:2014-01-03
  • Contact: LEI Sheng-jiao

摘要:

以戊二醛为交联剂,壳聚糖为载体,采用交联-吸附偶联法固定柚苷酶,通过单因素和正交试验优化确定最佳固定化工艺。结果表明,柚苷酶的最佳固定化条件为:以质量浓度为3.5g/100mL的壳聚糖制备的凝胶微球为载体,凝结剂NaOH质量浓度1.0g/100mL、戊二醛体积分数7.0%、交联时间2.0h、pH 4.0、酶液质量浓度2.0mg/mL、25℃时吸附交联3.0h,得到固定化酶最高酶比活力为7.37U/g;与游离酶相比而言,固定化酶最适pH值与最适反应温度均无明显变化;固定化酶在不同温度(40、50、60℃)条件下重复使用7次,相对酶活力仍能保持在70%、60%和50%以上。

关键词: 壳聚糖, 柚苷酶, 固定化

Abstract:

Naringinase (EC 3.2.1.40) was immobilized onto glutaraldehyde cross-linked chitosan beads by an adsorption
method. Using one-factor-at-a-time and orthogonal array designs, the optimal immobilization conditions were determined as
follows: gel microspheres prepared with 3.5 g/100 mL chitosan as the carrier, 1.0 g/100 mL NaOH as the coagulant, 7.0%
glutaraldehyde solution, 2.0 h of cross-linking, 2.0 mg/mL of naringinase concentration and 3.0 h of adsorption at 25 ℃.
The maximum specific activity of the immobilized naringinase under these conditions was 7.37 U/g (based on dry chitosan)
without showing obvious differences in the optimum pH and temperature compared to the free enzyme. After 7 cycles of
repeated use at 40, 50 ℃ and 60 ℃, the immobilized enzyme maintained above 70%, 60% and 50% of its original activity.

Key words: chitosan, naringinase, immobilization

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