双交联剂体系氨基酰化酶交联聚体的制备

摘要采用酶交联聚集体技术（CLEAs）制备氨基酰化酶(EC 3.51.14)交联聚集体，优化了制备条件。90%（体积比）乙醇为沉淀剂，戊二醛和乙二醇二缩水甘油醚（EGDE）作为混合交联剂，以酶、戊二醛和EGDE的质量比为1.00∶0.75∶2.00，在30 ℃下交联12 h。制备所得氨基酰化酶CLEA酶活保留率为63.42%。氨基酰化酶经固定化后热稳定性得到增强,热失活过程由自由酶的一阶指数失活模型变为二阶指数失活模型，分子间亚基的结合力得到显著加强。自由酶在37 ℃下保存24 h后几乎损失了所有活力，而氨基酰化酶CLEA则保留了43%的酶活力，在57 ℃下保存24 h后仍有32%的酶活力保留。此外，氨基酰化酶CLEA重复使用性能得到明显提升，在重复水解底物10次后仍保留了72%的酶活力。

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	余舜雷
	赵林
	董涛
	谭欣
	刘长光

关键词：酶交联聚集体氨基酰化酶酶活力

Abstract： The crosslinked enzyme aggregate (CLEA) of aminoacylase (EC 3.5, 1.14) from aspergillus melleus was prepared by precipitation with ethanol and crosslinking with glutaraidehyde and ethylene glycol diglycidyl ether (EGDE) as crosslinkers. The various crosslinked parameters were optimized. Almost 63.42% activity of CLEA of aminoacylase remained by using 90% (V/V) ethanol, followed by cross-linking 12 h with glutaraidehyde and EGDE at 30 ℃, the mass ratio was 1.00∶0.75∶2.00. The stability of aminoacylase was enhanced after immobilization. The thermal deactivation model was changed from oneexponential model (free enzyme) to twoexponential model. Free enzyme lost almost all of activity, while aminoacylase CLEA 43% of initial activity. Even at 57 ℃ after 24 h, 32% of initial activity was retained. Additionally, the reusability of the CLEA was evaluated. Aminoacylase CLEA showed 72% residual activity after 10 cycles of repeated use.

Keywords： cross-linked enzyme aggregate aminoacylase activity

Received 2012-05-06;

Corresponding Authors: 赵林，电话：18602612749，E-mail:zhaolin@tju.edu.cn。 Email: zhaolin@tju.edu.cn

About author: 余舜雷（1987-），男，硕士研究生，现从事酶固定化方面的研究。

引用本文:

余舜雷, 赵林, 董涛, 谭欣, 刘长光.双交联剂体系氨基酰化酶交联聚体的制备[J]. 化学工业与工程, 2013,30(3): 38-43

YU Shun-Lei, ZHAO Lin, DONG Tao, TAN Xin, LIU Chang-Guang.Preparation of CrossLinked Aggregates of Aminoacylase by Using Dual CrossLinkers[J]. Chemcial Industry and Engineering, 2013,30(3): 38-43