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›› 2010, Vol. 10 ›› Issue (4): 743-749.

• 生化工程专栏 • 上一篇    下一篇

静电纺丝制备苯乙烯马来酸酐共聚物纳米纤维及其固定化b-D-半乳糖苷酶

郑国建 张松平 张羽飞   

  1. 中国科学院过程工程研究所生化工程国家重点实验室 中国科学院过程工程研究所生化工程国家重点实验室 中国科学院过程工程研究所生化工程国家重点实验室
  • 收稿日期:2010-05-12 修回日期:2010-06-10 出版日期:2010-08-20 发布日期:2010-08-20
  • 通讯作者: 郑国建

Preparation of Electrospun Poly(styrene-co-maleic anhydride) Nanofibers and Their Use in Immobilization of b-D-Galactosidase

ZHENG Guo-jian, ZHANG Song-ping ZHANG Yu-fei,   

  1. 1State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, CAS National Key Laboratory of Biochemical Engingeering, Institute of Process Engineering, Chinese Academy of Sciences State Key Laboratory of Biochemical Engingeering, Institute of Process Engineering, Chinese Academy of Sciences
  • Received:2010-05-12 Revised:2010-06-10 Online:2010-08-20 Published:2010-08-20
  • Contact: ZHENG Guo-jian,

摘要: 采用静电纺丝技术制备苯乙烯-马来酸酐共聚物纳米纤维,最佳电纺条件为:聚合物浓度0.35 g/mL、针尖到接收板距离25 cm、电纺液流量250 mL/h、电压21 kV. 该条件下获得了直径约300 nm且分布均一的纳米纤维. 利用该纳米纤维固定b-D-半乳糖苷酶,固定化反应的最适pH值为4.0,此时酶负载量为(15.1±0.5) mg/g. 固定化酶催化2-硝基苯酚-b-D-半乳吡喃糖苷水解反应的米氏常数Km=2.7 mmol/L,略大于游离酶的Km值(2.2 mmol/L);最大反应速率Vmax为97.2 mmol/(min×mg),为游离酶的47.8%. 固定化酶在37℃下重复操作21次后活性损失仅为15%. 在连续搅拌式反应器中将固定化酶用于催化乳糖的水解反应,连续使用17 d仍能稳定运行.

关键词: 静电纺丝, 纳米纤维, 苯乙烯-马来酸酐共聚物, b-D-半乳糖苷酶, 乳糖水解

Abstract: Unique poly(styrene-co-maleic anhydride) nanofibers with a uniform diameter of about 300 nm were prepared by eletrospinning a solution containing 0.35 g/mL polymer, 25 cm gap between ejection needle tip and collector, flow rate of 250 L/h, and applied voltage of 21 kV. The electrospun nanofibers were applied subsequently for immobilization of b-D-galactosidase, an enzyme loading of (15.1±0.5) mg protein/g membrane was obtained. The immobilized enzyme demonstrated a Michealis constant (Km) of 2.7 mmol/L, slightly higher than that of the native enzyme (2.2 mmol/L). The maximum reaction velocity (Vmax) of the immobilized enzyme was found as 97.2 mol/(min×mg), 47.8% that of native enzyme. The immobilized enzyme showed an excellent reusability such that 21 reusing cycles only led to 15% loss of the enzyme activity. Furthermore, the immobilized b-D-galactosidase was suitable for hydrolysis of lactose in a continuous stirring tank reactor that was operated steadily for 17 d.

Key words: electrospinning, nanofiber, poly(styrene-co-maleic anhydride), b-D-galactosidase, lactose hydrolysis

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