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CS-Cu酶膜的酶固定化工艺及其特性

作者：葛振，蔡邦肖¹，²

单位： 1.浙江工商大学食品与生物工程学院膜科学与工程研究所, 浙江杭州 310035;

关键词：酶膜反应器; 固定化酶; 壳聚糖铜膜; 多酚氧化酶; 响应面

DOI号：

分类号： TQ028.8,R978.1

出版年,卷(期):页码： 2012,32(1):64-70

摘要：

固定多酚氧化酶（PPO）的酶膜反应器（EMR）由于其优良的分离与生化特性而受到广泛关注。本文以壳聚糖（CS）膜为载体，通过PPO与CS-Cu膜上的Cu2+鳌合，制备固定化PPO的CS-Cu酶膜。响应面法研究了酶膜在固定化过程中的Cu2+螯合量（QCu2+）、PPO浓度（CE）、pH三因素对固定化PPO的活力的影响，分别测定了溶液PPO和酶膜上固定化PPO的酶活性，并对CS-Cu–PPO膜的特性作了初步讨论。结果表明，PPO固定化的最优条件是：温度40℃，pH 7.67，QCu2+ 30.07 mg/mL，CE 2.12 mg/mL；酶膜中固定化PPO的米氏常数及其相对活力都比溶液PPO的大，半衰期约30d，酶膜重复使用8次后的PPO活力降至79%。研究结果为开发特定功能的EMR提供了重要的基础。

Enzymatic membrane reactor(EMR) with immobilization of polyphenol oxidase(PPO) is paid more attention due to its isolation and biochemical characteristics. In this paper, a enzymatic membrane immobilized PPO was prepared based on the chelation of PPO and Cu2+ in chitosan(CS)-Cu complex(CCC) with CS membrane as the carrier. The response surface methodology was applied to investigate the influence of three factors of copper ion sequestration, QCu2+, PPO concentration, CE, pH in immobilization process on the vitality of PPO immobilized in CCC. The relative activity of PPO in the solution and immobilized in the membrane was determined respectively. The characteristics of CS-Cu–PPO membrane were discussed primarily. The results show that the optimal conditions for immobilization of PPO into the CCC were temperature 40℃, pH 7.67, QCu2+ 30.07 mg/mL, CE 2.12 mg/mL. Michaelis constant and the relative activity of PPO immobilized in CCC were larger than that of the solution. Enzyme half-decline period was 30 days, and the relative activity of PPO after repeated use of eight times reduced to 79% for CS-Cu–PPO membrane. This work offers the significantly basis for development of EMR with special function.

基金项目：

国家科技支撑计划项目(2007BAD62B05), 国家农业部行业科技开发专项(nyhyzx07-21)

作者简介：

葛振(1985-), 男, 山东人, 食品科学硕士研究生，研究方向：新型分离膜及其应用工程. *通讯联系人: 蔡邦肖, 教授. 13958002662, caibangxiao@163.com

参考文献：

[1] S. T. Bouwer, F. P. Cuperus, and J. T. P. Derksen. The performance of enzyme-membrane reactors with immobilized lipase[J]. Enzyme and Microbial Technology, 1997, 21(1):291-296.
[2] G. M. Rios, M. P. Belleville, D. Paolucci, et. al. Progress in enzymatic membrane reactors – a review[J]. Journal Membrane Science, 2004, 242(1-2):189-196.
[3] L. J. Kennedy, P. K. Selvi, A. Padmanabhan, et al. Immobilization of polyphenol oxidase onto
mesoporous activated carbons – isotherm and kinetic studies [J]. Chemosphere, 2007, 69 (2):
262-270.
[4] S. Kiralp, L.Toppare, Y.Yagci. Immobilization of polyphenol oxidase in conducting copolymers
and determination of phenolic compounds in wines with enzyme electrodes[J]. International
Journal of Biological Macromolecules, 2003, 33 (1-3): 37-41.
[5] I. Mielgo, C. Palma, J. M. Guisan, et al. Covalent immobilization of manganese peroxidases (MnP) from Phanerochaete chrysosporium and Bjerkandera sp. BOS55[J]. Enzyme and Microbial Technology, 2003, 32 (7): 769-775.
[6] T. Gumi, D. Paolucci-Jeanjean, B. Marie-Pierre, et al. Enzymatic membrane reactor involving a hybrid membrane in supercritical carbon dioxide[J]. Journal of Membrane Science, 2007, 297(1-2): 98-103.
[7] B. Krajewska. Application of chitin- and chitosan-based materials for enzyme immobilizations: a
review [J]. Enzyme and Microbial Technology, 2004, 2(35): 126-139.
[8] N. Duran, E. Esposito. Potential application of oxidative enzymes and phenoloxidase-like compounds in wastewater and soil treatment: a review[J]. Applied Catalysis B: Environmental, 2000, 28 (2): 83-99.
[9] J. Z. Lu, X. F. Duan, Q. L. Wu, et al. Chelating e?ciency and thermal, mechanical and decay resistance performances of chitosan copper complex in wood–polymer composites[J]. Bioresource Technology, 2008, 99 (13 ) :5906-5914.
[10] 姜梅, 王善荣, 季勤. 壳聚糖膜固定化葡萄糖氧化酶的特性研究[J]. 食品科学, 2003, 24(4): 38-40.
[11] 黄意欢. 茶学实验技术 [M]. 北京:中国农业出版社, 1995: 213-214.
[12] 雷福厚. 壳聚糖铜固定化多酚氧化酶的研究 [J]. 林产化学与工业, 2000, 2 (20): 23-26.
[13] 罗贵民. 酶工程 [M] . 北京:化学工业出版社, 2003 : 26-27.
[14] S. Vishwanath, D. Bhattacharyya, W. Huang, et al. Site-directed and random enzyme immobilization on functionalized membranes: kinetic studies and models[J]. Journal of Membrane Science, 1995, 108 (2): 1-13.

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