介孔二氧化钛固载γ-谷氨酰转肽酶的制备及性质

摘要/Abstract

摘要：

γ-谷氨酰转肽酶（GGT）在临床诊断和生物催化方面具有重要的应用价值。本文以介孔氧化钛晶须为载体进行GGT的固定化，考察了载体结构特性、吸附时间和给酶量对固定化效果的影响，并对固定化酶的催化特性及其稳定性进行了研究。结果显示，以最可几孔径为30 nm的介孔TiO₂为载体，载体载酶量可达5.07mg·g^-1。在给酶量为18.99 U·g^-1时，经室温吸附2.5 h，固定化酶活性回收率可达73.05%。固定化酶的pH稳定性和热稳定性均显著优于游离酶，在4℃下保温贮藏60 d、转化22个批次后，固定化酶活力仍可保持初始值的71.30%。经测定，游离酶和固定化酶的米氏常数K_m分别为0.79 mmol·L^-1和1.05 mmol·L^-1，酰基化反应活化能分别为13.59 kJ·mol^-1和15.42 kJ·mol^-1；固定化GGT的失活反应活化能E_d为92.80 kJ·mol-1，相比于游离酶（49.61 kJ·mol^-1）有明显的增加。

关键词: γFONT-FAMILY: Calibri,sans-serif, mso-bidi-font-size: 11.0pt, mso-bidi-font-family: 宋体, mso-font-kerning: 1.0pt, mso-ansi-language: EN-US, mso-fareast-language: ZH-CN, mso-bidi-language: AR-SA, mso-fareast-font-family: 宋体, mso-hansi-font-family: 宋体">-FONT-FAMILY: 宋体, mso-bidi-font-size: 11.0pt, mso-ascii-font-family: Calibri, mso-bidi-font-family: 宋体, mso-font-kerning: 1.0pt, mso-ansi-language: EN-US, mso-fareast-language: ZH-CN, mso-bidi-language: AR-SA">谷氨酰转肽酶；固定化；介孔二氧化钛；酶学性质

Abstract:

γ-Glutamyltranspeptidase (GGT）is an important enzyme with wide applications in biocatalysis and clinical diagnosis.In this work, mesoporous fibrous titania (M-TiO₂）was used for immobilization of GGT from B.subtilis NX-2 and the properties of immobilized GGT were also investigated.When the M-TiO₂support with average pore diameter of 30 nm was used, the amount of immobilized protein was 5.07 mg·g^-1, and the yield of activity was 73.05% at the ratio of GGT/support was 18.99 U·g^-1 after incubation at room temperature for 2.5 h.The thermal and pH stability of the immobilized GGT was higher than that in its free form.After storage at 4℃ for 60 days and repeated use for 22 batches, the activity of the immobilized GGT remained 71.30% of its initial activity.The kinetic parameters (K_m）for free and immobilized GGT were determined as 0.79 mmol·L^-1 and 1.05 mmol·L^-1, respectively.The activation energy (E_a）values of glutamylation were 15.42 kJ·mol^-1^and 13.59 kJ·mol^-1 for immobilized and free GGT.The thermal inactivation energy (E_d）values of GGT for immobilized and free enzyme were also calculated to be 92.80 kJ·mol^-1 and 49.61 kJ·mol^-1, respectively.

Key words: γFONT-FAMILY: Calibri,sans-serif, mso-bidi-font-size: 11.0pt, mso-bidi-font-family: 宋体, mso-font-kerning: 1.0pt, mso-ansi-language: EN-US, mso-fareast-language: ZH-CN, mso-bidi-language: AR-SA, mso-fareast-font-family: 宋体, mso-hansi-font-family: 宋体">-FONT-FAMILY: 宋体, mso-bidi-font-size: 11.0pt, mso-ascii-font-family: Calibri, mso-bidi-font-family: 宋体, mso-font-kerning: 1.0pt, mso-ansi-language: EN-US, mso-fareast-language: ZH-CN, mso-bidi-language: AR-SA">谷氨酰转肽酶；固定化；介孔二氧化钛；酶学性质

秦丹华, 姚忠, 王浩琦, 肖易凡, 杨敬, 徐虹, 韦萍. 介孔二氧化钛固载γ-谷氨酰转肽酶的制备及性质 [J]. 化工学报, 2011, 62(2): 378-385.

QIN Danhua, YAO Zhong, WANG Haoqi, XIAO Yifan, YANG Jing, XU Hong, WEI Ping. mmobilization of GGT on meso-TiO₂: reparation and properties[J]. CIESC Journal, 2011, 62(2): 378-385.

[1]	胡朝发, 贾力. 脉动热管气液塞振荡运动模型 [J]. 化工学报, 2011, 62(S1): 113-117.
[2]	李旺, 王情愿, 李瑞龙, 李超, 宇波, 张劲军, 代鹏飞. 大型浮顶油罐温度场数值模拟 [J]. 化工学报, 2011, 62(S1): 108-112.
[3]	刘绩伟, 吴玉庭, 马重芳. 管肋式空间辐射器设计 [J]. 化工学报, 2011, 62(S1): 103-107.
[4]	何奎, 汪双凤, 黄间珍. 气液两相流在微小T型三通中的相分配特性 [J]. 化工学报, 2011, 62(S1): 92-96.
[5]	崔福庆, 何雅玲, 程泽东, 李东, 陶于兵. 有压腔式吸热器内辐射传播过程的Monte Carlo模拟 [J]. 化工学报, 2011, 62(S1): 60-65.
[6]	马维刚, 王海东, 张兴, 王玮. 多晶金薄膜电子-声子耦合的实验研究 [J]. 化工学报, 2011, 62(S1): 48-53.
[7]	王建立, 朱建军, 宋辰兴, 张兴. 3ω法测量纳米流体热导率 [J]. 化工学报, 2011, 62(S1): 42-47.
[8]	任楠, 王涛, 吴玉庭, 马重芳. 混合碳酸盐的DSC测量与比热容分析 [J]. 化工学报, 2011, 62(S1): 197-202.
[9]	褚燕彬, 曹柳林, 王晶. 利用组合B样条神经网络实现对间歇反应过程的动态建模 [J]. 化工学报, 2011, 62(S1): 157-162.
[10]	曾庆, 郭印诚, 牛振祺, 林文漪. 填料塔中氨水吸收二氧化碳的传质性能 [J]. 化工学报, 2011, 62(S1): 146-150.
[11]	张艾萍, 谢立强, 徐志明. 超声空化对换热器协同场的影响 [J]. 化工学报, 2011, 62(S1): 130-133.
[12]	吴海波, 方岩雄, 纪红兵. 助溶剂促进的β-环糊精与肉桂醛的包结性能 [J]. 化工学报, 2011, 62(S1): 168-173.
[13]	王勤辉，杨秋辉，吴学成，骆仲泱，岑可法. 多相流中颗粒旋转运动特性的研究进展 [J]. CIESC Journal, 2011, 62(9): 2381-2390.
[14]	刘夺, 杜瑾, 赵广荣, 元英进. 合成生物学在医药及能源领域的应用 [J]. 化工学报, 2011, 62(9): 2391-2397.
[15]	马淳安，储诚普，徐颖华，张虹. 3,6-二氯吡啶甲酸合成方法进展 [J]. CIESC Journal, 2011, 62(9): 2398-2405.