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›› 2010, Vol. 10 ›› Issue (6): 1181-1186.

• 生化工程专栏 • 上一篇    下一篇

咪唑和吡啶离子液体对青霉素酰化酶催化性能的影响

周华从 李伟 高红帅 贾连伟 徐芃 刘会洲   

  1. 中国科学院过程工程研究所绿色过程与工程实验室 中国科学院过程工程研究所绿色过程与工程实验室 中国科学院过程工程研究所绿色过程与工程实验室 中国科学院过程工程研究所绿色过程与工程实验室 中国科学院过程工程研究所分离工程与工程青年实验室
  • 收稿日期:2010-09-17 修回日期:2010-11-15 出版日期:2010-12-20 发布日期:2010-12-20
  • 通讯作者: 周华从

Effects of Imidazole- and Pyridine-based Ionic Liquids on Catalysis Performance of Immobilized Penicillin Acylase

ZHOU Hua-cong LI Wei GAO Hong-shuai JIA Lian-wei XU Peng LIU Hui-zhou   

  1. Laboratory of Green Process and Engineering, Institute of Process Engineering, CAS Laboratory of Green Process and Engineering, Institute of Process Engineering, CAS Laboratory of Green Process and Engineering, Institute of Process Engineering, CAS Laboratory of Green Process and Engineering, Institute of Process Engineering, CAS Institute of Process Engineering, Chinese Academy of Sciences
  • Received:2010-09-17 Revised:2010-11-15 Online:2010-12-20 Published:2010-12-20
  • Contact: ZHOU Hua-cong

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摘要： 采用两步法合成并表征了阳离子不同的咪唑型和吡啶型离子液体[C4mim]BF4, [C7mim]BF4, [C4pyr]BF4和[C7pyr]BF4，并将其应用于固定化青霉素G酰化酶(ImPGA)催化分解青霉素G钾盐的反应，考察了阳离子对酶活性、稳定性、动力学的影响，及ImPGA在不同离子液体中酶活性受底物(青霉素G钾盐)抑制情况. 结果表明，ImPGA在[C7pyr]BF4, [C4pyr]BF4和[C7mim]BF4中活性依次降低，在亲水性较强的[C4mim]BF4中易失去活性，其在3种离子液体中37℃下保存30 min活性没有降低，米氏常数Km依次为752.15, 160.57和62.74 mmol/L，最大反应速率Vmax依次为0.204, 0.042和0.024 mmol/(g×min)；底物浓度为26.85~295.32 mmol/L时酶活性不受底物抑制.

关键词: 青霉素酰化酶, 离子液体, 酶活性, 稳定性, 动力学

Abstract: In order to investigate the effect of cations of ionic liquids on catalysis performance of immobilized penicillin G acylase (ImPGA). Four kinds of imidazole- and pyridine-based ionic liquids through two-step strategy, [C4mim]BF4, [C7mim]BF4, [C4pyr]BF4 and [C7pyr]BF4 were synthesized and characterized. The hydrolysis of penicillin G potassium catalyzed by ImPGA was conducted in these ionic liquids. Enzyme activity, stability and kinetics of ImPGA were studied. Substrate inhibition of ImPGA under different substrate concentrations was also detected. The results showed that the activity of ImPGA decreased in the order of [C7pyr]BF4>[C4pyr]BF4>[C7mim]BF4. ImPGA lost its activity in [C4mim]BF4 because of high hydrophilicity of the media. Enzyme activity of ImPGA could be maintained after incubation in ionic liquids at 37℃ for 30 min in the former three ionic liquids. Two kinetic parameters, Km and Vmax, of the reaction decreased followed the order [C7pyr]BF4>[C4pyr]BF4>[C7mim]BF4. Enzyme activity was not inhibited by substrate at the concentration of 75.17~85.91 mmol/L.

Key words: penicillin acylase, ionic liquids, enzyme activity, stability, kinetics

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