基于多级孔金属有机骨架构筑HRP固定化酶反应器及其染料降解应用

doi:10.7503/cjcu20200130

高等学校化学学报 ›› 2020, Vol. 41 ›› Issue (7): 1591.doi: 10.7503/cjcu20200130

基于多级孔金属有机骨架构筑HRP固定化酶反应器及其染料降解应用

高霞^1,^2,^*(),潘会宾³,乔成芳^1,^*(),陈凤英¹,周元¹,杨文华¹

1.商洛学院化学工程与现代材料学院, 陕西省尾矿资源综合利用重点实验室, 商洛 726000
2.陕西师范大学化学化工学院, 大分子科学陕西省重点实验室, 西安 710119
3.商洛职业技术学院公共基础教学部, 商洛726000

收稿日期:2020-03-09 出版日期:2020-07-10 发布日期:2020-04-15
通讯作者: 高霞,乔成芳 E-mail:xiagao2007@163.com;xiaoqiaoqcf@126.com
基金资助:
国家自然科学基金(21703135);陕西省教育厅自然科学基金(17JS034);商洛学院自然科学计划项目(19SKY003)

Construction of HRP Immobilized Enzyme Reactor Based on Hierarchically Porous Metal-organic Framework and Its Dye Degradation Application^†

GAO Xia^1,^2,^*(),PAN Huibin³,QIAO Chengfang^1,^*(),CHEN Fengying¹,ZHOU Yuan¹,YANG Wenhua¹

1. Shaanxi Key Laboratory of Comprehensive Utilization of Tailings Resources, Department of Chemical Engineering & Modern Materials, Shangluo University, Shangluo 726000, China
2. Key Laboratory of Macromolecular Science of Shaanxi Province, School of Chemistry & Chemical Engineering, Shaanxi Normal University, Xi’an 710119, China
3. Public Basic Teaching Division, Shangluo Vocational & Technical College, Shangluo 726000, China

Received:2020-03-09 Online:2020-07-10 Published:2020-04-15
Contact: Xia GAO,Chengfang QIAO E-mail:xiagao2007@163.com;xiaoqiaoqcf@126.com
Supported by:
† National Natural Science Foundation of China(21703135);Natural Science Foundation of the Department of Education of Shaanxi Province, China(17JS034);Natural Science Program Funded by Shangluo University, China(19SKY003)

摘要/Abstract

摘要：

以不稳定的Cu-金属有机骨架(Cu-MOF)为模板剂, 利用自组装模板法制备多级孔Zr-MOF, 再通过物理吸附法在多级孔Zr-MOF的介孔孔道中负载辣根过氧化物酶(HRP)构筑了HRP@Zr-MOF固定化酶反应器. 通过改变孔径调节剂苯甲酸(HBC)的浓度调控孔径大小, 研究了孔径对固定化酶反应器催化活性的影响; 考察了固定化体系缓冲溶液pH值、固定化时间及温度对固定效果的影响. 以HRP催化降解结晶紫染料为模型反应, 探讨了HRP@Zr-MOF的操作稳定性和重复使用性. 结果表明, pH=3.0、固定化时间为60 min、固定化温度为30 ℃是固定化HRP的最佳条件, 固载量最高可达61.6 mg/g. 与游离酶相比, HRP@Zr-MOF固定化酶反应器表现出更好的热稳定性、酸碱稳定性、 H₂O₂稳定性和储存稳定性; 重复使用10次后, HRP@Zr-MOF的催化活性仍能保持62.3%. 将HRP@Zr-MOF应用于实际水样中结晶紫染料的催化降解, 在5 min内降解率高达90%以上, 表现出非常高效的催化效率.

关键词: 金属有机骨架, 辣根过氧化物酶, 固定化酶反应器, 染料降解

Abstract:

Using the unstable Cu-MOF as template agent, the hierarchically porous Zr-MOF was prepared by a self-assembled template method. Horseradish peroxidase(HRP) was successfully immobilized into the hierarchically porous Zr-MOF via physical adsorption. The effect of pore size on catalytic activity of immobilized enzyme reactor was studied by changing the concentration of the pore size regulator benzoic acid(HBC). The effects of pH value of buffer solution, immobilized time and temperature on immobilized effect were investigated. The HRP-catalyzed degradation of crystal violet was used as a model reaction to study the operational stability and reusability of HRP@Zr-MOF. The results showed that the optimal conditions are the buffer pH=3.0, the immobilization time of 60 min and the temperature of 30 ℃, and the maximum immobilization was 61.6 mg/g. HRP@Zr-MOF showed better operational stability than the free enzyme. After 10 cycles, HRP@Zr-MOF can still remain 62.3% of the catalytic activity. When HRP@Zr-MOF was applied to the degradation of crystal violet in real water samples, they show very high catalytic efficiency with the degradation rate of more than 90% within 5 min.

Key words: Metal-organic framework(MOF), Horseradish peroxidase(HRP), Immobilized enzyme reactor, Dye degradation

中图分类号:

TrendMD:

高霞,潘会宾,乔成芳,陈凤英,周元,杨文华. 基于多级孔金属有机骨架构筑HRP固定化酶反应器及其染料降解应用. 高等学校化学学报, 2020, 41(7): 1591.

GAO Xia,PAN Huibin,QIAO Chengfang,CHEN Fengying,ZHOU Yuan,YANG Wenhua. Construction of HRP Immobilized Enzyme Reactor Based on Hierarchically Porous Metal-organic Framework and Its Dye Degradation Application^†. Chem. J. Chinese Universities, 2020, 41(7): 1591.

图/表 14

Table 1 Pore sizes of Zr-MOF and catalytic activity of HRP@Zr-MOF

Carrier	n(HBC)/mmol	Catalytic activity(%)	Hierarchically porous size/nm	Adsorption average pore diameter/nm
Zr-MOF-1	5	56.88	1.4/2.8/4.5	4.2
Zr-MOF-2	10	98.32	1.5/2.8/8.8	8.6
Zr-MOF-3	15	90.35	1.3/11.8/18.5	12.8

Fig.1 Effects of the pH values(A), time(B) and temperatures(C) on the immobilization of HRP with Zr-MOF

Fig.2 N2 adsorption-desorption isotherms(A) and pore size distributions(B) of Zr-MOF

Fig.3 PXRD patterns of Zr-MOF(a), Zr-MOF after activation(b) and simulate UiO-66(c)(A) and PXRD patterns of Zr-MOF after activation(a) and HRP@Zr-MOF(b)(B)

Fig.4 FESEM images of Zr-MOF(A) and HRP@Zr-MOF(B)

Fig.5 TEM images of Zr-MOF(A) and HRP@Zr-MOF(B)

Fig.6 Laser confocal microscopy images of the Zr-MOF(A) and FITC-HRP@Zr-MOF(B,C)

Fig.7 UV-Vis absorbance spectra of oxidation degradation of crystal violet catalyzed by free HRP and HRP@Zr-MOF

Fig.8 Temperature stability of free HRP and HRP@Zr-MOF

Fig.9 Effects of pH values on the catalytic activity of free HRP and HRP@Zr-MOF

Fig.10 Effects of H2O2 concentration on the catalytic activity of free HRP and HRP@Zr-MOF

Fig.11 Storage stability of free HRP and HRP@Zr-MOF

Fig.12 Operational reusability of HRP@Zr-MOF

Fig.13 Decolorization of crystal violet with HRP@Zr-MOF in actual wastewater

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基于多级孔金属有机骨架构筑HRP固定化酶反应器及其染料降解应用

Construction of HRP Immobilized Enzyme Reactor Based on Hierarchically Porous Metal-organic Framework and Its Dye Degradation Application^†

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基于多级孔金属有机骨架构筑HRP固定化酶反应器及其染料降解应用

Construction of HRP Immobilized Enzyme Reactor Based on Hierarchically Porous Metal-organic Framework and Its Dye Degradation Application†

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Construction of HRP Immobilized Enzyme Reactor Based on Hierarchically Porous Metal-organic Framework and Its Dye Degradation Application^†