氮掺杂中空碳球氧化物模拟酶性能研究

doi:10.15541/jim20200396

无机材料学报 ›› 2021, Vol. 36 ›› Issue (5): 527-534.DOI: 10.15541/jim20200396

氮掺杂中空碳球氧化物模拟酶性能研究

郑燕宁¹^,²(), 季军荣³, 梁雪玲¹, 赖正杰¹, 陈启帆¹, 廖丹葵¹^,²

1.广西大学化学化工学院, 南宁 530004
2.广西大学广西碳酸钙产业化工程院, 广西钙基材料创新协同中心, 南宁 530004
3.崇左南方水泥有限公司广西钙基材料创新协同中心, 崇左 532201

收稿日期:2020-07-04 修回日期:2020-08-20 出版日期:2021-05-20 网络出版日期:2021-04-19
通讯作者: 廖丹葵, 教授. E-mail: liaodk@gxu.edu.cn
作者简介:郑燕宁(1995-), 女, 硕士研究生. E-mail:ningningzy@sina.cn
基金资助:
国家自然科学基金(51372043);广西自然科学基金(2017GXNSFDA198052);广西大学钙基材料协同创新研究项目(20190962)

Performance of Nitrogen-doped Hollow Carbon Spheres as Oxidase Mimic

ZHENG Yanning¹^,²(), JI Junrong³, LIANG Xueling¹, LAI Zhengjie¹, CHENG Qifan¹, LIAO Dankui¹^,²

1. School of Chemistry and Chemical Engineering, Guangxi University, Nanning 530004, China
2. Guangxi Cooperative Innovation Centre for Calcium-based Materials (GCICCM), Guangxi Engineering Academy for Calcium Carbonate Industry, Guangxi University, Nanning 530004, China
3. Guangxi Cooperative Innovation Centre for Calcium-based Materials (GCICCM), Chongzuo South Cement Co. LTD, Chongzuo 532201, China

Received:2020-07-04 Revised:2020-08-20 Published:2021-05-20 Online:2021-04-19
Contact: LIAO Dankui, professor.E-mail: liaodk@gxu.edu.cn
About author:ZHENG Yanning(1995-), female, Master candidate. E-mail:ningningzy@sina.cn
Supported by:
National Natural Science Foundation of China(51372043);Foundation of Guangxi Natural Science(2017GXNSFDA198052);Research Project on Collaborative Innovation of Calcium-based Materials in Guangxi University(20190962)

摘要/Abstract

摘要：

纳米酶由于其独特、高效、稳定的催化性质而在生化反应中备受关注。本研究以碳酸钙微球为绿色模板剂, 多巴胺为氮源与碳源, 合成了氮掺杂中空碳球(N-HCSs)。以3,3°,5,5°-四甲基联苯胺(TMB)为底物, 采用紫外分光光度法探究了N-HCSs的类氧化物酶的催化活性, 并研究其催化机理。结果表明, N-HCSs具有氧化物模拟酶催化活性, KOH活化后氮掺杂中空碳球的催化活性提高了3倍; N-HCSs氧化物模拟酶催化反应符合Michaelis-Menten方程, 活化前后的米氏常数K_m分别为0.105和0.083, 对底物具有良好的亲和能力; N-HCSs氧化物模拟酶催化反应中起主要作用的活性氧基团是超氧阴离子(O₂ ^?-)。本研究为高活性无机非金属类氧化物模拟酶的设计和制备提供了理论依据。

关键词: 氮掺杂中空碳球, 氧化物模拟酶, 反应机理

Abstract:

Due to efficient performance and stability, nanozymes have recently attracted much attention in bioreaction. In this work, a facile approach for preparing N-doped hollow carbon sheres (N-HCSs) by using CaCO₃ spheres as green template and polydoamine as nitrogen and carbon sources was reported. Morphologies and structures of the samples were characterized. Using TMB (3,3,5,5° tetramethylbenzidine) as a substrate, UV spectra photometry was used to investigate its oxidase-like activity and catalytic mechanism. The results showed that N-HCSs displayed oxidase-like activity. The oxidase-like activity of N-HCSs increased by three times after activation by KOH. These enzymes conform to the Michaelis-Menten kinetic equation, and the K_m constant before and after activation were 0.105 and 0.083, respectively, indicating good affinity for substrates. The data demonstrate that it is superoxide anion (O₂? ^-) that plays a major role in catalytic reaction. This work data provides a theoretical basis for the design and preparation of high activity oxidase-mimicking enzymes.

Key words: N-doped hollow carbon sheres, oxidase-like enzyme, reaction mechanism

中图分类号:

TQ174

郑燕宁, 季军荣, 梁雪玲, 赖正杰, 陈启帆, 廖丹葵. 氮掺杂中空碳球氧化物模拟酶性能研究[J]. 无机材料学报, 2021, 36(5): 527-534.

ZHENG Yanning, JI Junrong, LIANG Xueling, LAI Zhengjie, CHENG Qifan, LIAO Dankui. Performance of Nitrogen-doped Hollow Carbon Spheres as Oxidase Mimic[J]. Journal of Inorganic Materials, 2021, 36(5): 527-534.

图/表 10

图1 N-HCSs的形貌和元素分析 (a) Schematic presentation for the oxide enzyme mimetic activity of N-HCSs; (b) STEM image of N-HCS; (c) C, (d) N and (e) O EDX mappings of N-HCS

Fig. 1 Morphologies and element mappings of N-HCSs

图2 (a)N-HCS和(b)N-HCS-1的SEM照片; (c)CaCO3@PDA和(d)N-HCS的TEM照片

Fig. 2 SEM images of (a) N-HCS and (b) N-HCS-1, and TEM images of (c) CaCO3@PDA and (d) N-HCS

图3 (a)N-HCS和N-HCS-1的XPS全谱, (b)N-HCS和(c)N-HCS-1的N1s谱, (d)N-HCS和N-HCS-1的N元素组成

Fig. 3 (a) XPS full spectra of N-HCS and N-HCS-1; N1s spectra of (b) N-HCS and (c) N-HCS-1; (d) N species contents of N-HCS and N-HCS-1

图4 (a)N-HCS和N-HCS-1的N2吸附-脱附等温曲线及(b)HK模型孔径分布图

Fig. 4 (a) Nitrogen absorption-desorption isotherm curves of N-HCS and N-HCS-1, (b) HK model corresponding pore size distribution curves of N-HCS and N-HCS-1

表1 N-HCS和N-HCS-1材料的孔结构参数

Table 1 Pore structure parameters of N-HCS and N-HCS-1

Sample	Specfic suface/(m²•g^-1)		V_t/ (cm³•g^-1)	V_m/ (cm³•g^-1)	S_m/ (m²•g^-1)	(S_m/S_t)/ %
Sample	Langmuir	BET	V_t/ (cm³•g^-1)	V_m/ (cm³•g^-1)	S_m/ (m²•g^-1)	(S_m/S_t)/ %
N-HCS	685.6	490.1	0.651	0.109	230.51	45.49
N-HCS-1	730.0	518.4	0.732	0.111	228.16	44.01

图5 (a)不同体系的紫外-可见吸收光谱, (b)催化剂浓度、(c)pH和(d)温度对N-HCSs氧化物模拟酶活性的影响

Fig. 5 (a) UV-Vis absorption spectra of different systems, and effects of (b) catalyst concentration, (c) pH and (d) temperature on the oxidase-like activity of N-HCSs

图6 N-HCS和N-HCS-1稳态动力学分析

Fig. 6 Steady-state kinetic assay of N-HCS and N-HCS-1

表2 N-HCSs与其它氧化物模拟酶动力学参数的比较

Table 2 Comparison of the Km and Vmax between N-HCSs and other oxidase-like

Catalyst	Substrate	K_m/ (mmol•L^-1)	V_max/(×10^-8, mol•L^-1•s^-1)	Ref.
N-PCS	TMB	0.095	5.20	[4]
His@AuNCs	TMB	0.041	6.21	[31]
Pt Au DNPs	TMB	0.22	2.82	[32]
Acr⁺-Mes	TMB	0.129	2.68	[33]
N-HCS	TMB	0.1049	4.69	This work
N-HCS-1	TMB	0.0825	5.98	This work

图7 在体系中(a)通入氮气后的吸收光图谱, 及加入不同活性氧清除剂(b)异丙醇(IPA)、(c)过氧化氢酶(CAT)、(d)超氧化物歧化酶(SOD)对N-HCS和N-HCS-1类氧化物酶活性的影响

Fig. 7 (a) Absorption spectra of the solution containing TMB and N-HCS and N-HCS-1 under N2-saturated conditions, and effect of scavengers (b) IPA, (c) CAT and (d) SOD on the catalytic oxidation of TMB by the N-HCS and N-HCS-1

图8 (a)N-HCSs氧化物模拟酶的ESR图谱及其(b)催化机理图

Fig. 8 (a) ESR spectra and (b) catalytic mechanism diagram for the oxidase-like activity of the N-HCSs

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氮掺杂中空碳球氧化物模拟酶性能研究

Performance of Nitrogen-doped Hollow Carbon Spheres as Oxidase Mimic

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