基于硼酸-二醇特异性识别作用的层层组装薄膜对肌红蛋白的吸入及其电化学研究

doi:10.7503/cjcu20150943

摘要/Abstract

摘要：

合成了既含有苯硼酸(PBA)基团又含有羧酸基团的聚电解质PAA-PBA(PAA: 聚丙烯酸). 采用层层组装(LbL)技术, 利用PAA-PBA和葡聚糖(Dex)之间的硼酸-二醇特异性识别作用, 在热解石墨电极(PG)表面构筑了{PAA-PBA/Dex}_nLbL薄膜, 该薄膜从溶液中吸入肌红蛋白(Mb)形成{PAA-PBA/Dex}_n-Mb薄膜. 采用循环伏安方法研究了{PAA-PBA/Dex}_n薄膜中Mb的直接电化学及对氧气和过氧化氢的电催化还原过程. 结果表明, 该薄膜为保持Mb的生物活性提供了良好的微环境, 是一种新型的可固定蛋白质的LbL薄膜, 为设计基于酶的直接电化学生物传感器提供了新思路.

关键词: 直接电化学, 电催化, 层层组装, 硼酸-二醇特异性识别作用, 肌红蛋白

Abstract:

A polyelectrolyte PAA-PBA containing both phenylboronic acid(PBA) moieties and carboxylic acid groups[PAA=poly(acrylic acid)] was synthesized. PAA-PBA and dextran(Dex) were then assembled into {PAA-PBA/Dex}_n layer-by-layer(LbL) films by LbL assemble technique on pyrolytic graphite(PG) electrode surface through boronic acid-diol specific recognition between them. Myoglobin(Mb) in a buffer solution(pH=5.0) was loaded into the films, forming stable {PAA-PBA/Dex}_n-Mb films. By cyclic voltammetry(CV) technique, the {PAA-PBA/Dex}_n-Mb film electrodes demonstrated the direct electrochemistry of Mb and could be used to electrocatalyze reduction of various substrates(e.g. O₂ and H₂O₂).

Key words: Direct electrochemistry, Electrocatalysis, Layer-by-layer assembly, Boronic acid-diol specific recognition, Myoglobin

中图分类号:

O646
O657.1

TrendMD:

姚惠琴, 黄珊, 苏巧玲, 史可人, 甘倩倩, 王明科. 基于硼酸-二醇特异性识别作用的层层组装薄膜对肌红蛋白的吸入及其电化学研究. 高等学校化学学报, 2016, 37(5): 938.

YAO Huiqin, HUANG Shan, SU Qiaoling, SHI Keren, GAN Qianqian, WANG Mingke. Loading of Myoglobin into Layer-by-layer Films Assembled Through Boronic Acid-diol Specific Recognition and Its Electrochemical Study^†. Chem. J. Chinese Universities, 2016, 37(5): 938.

图/表 11

Scheme 1 Synthetic route of PAA-PBA

Fig.1 CVs of 1.0 mmol/L Fe(CN)63- at a scan rate of 0.1 V/s in pH=7.0 buffers at bare PG electrode(a), PG/CS film electrode(b) and PG/CS/{PAA-PBA/Dex}n film electrodes with n=1—7(c—h)

Fig.2 EIS responses of bare PG(a), PG/CS film(b), PG/CS {PAA-PBA/Dex}6 film(c) and PG/CS{PAA-PBA/Dex}6-Mb film(d) at 0.17 V in pH=7.0 buffers containing 5.0 mmol/L Fe(CN)3-/4-(A) and magnification of curves a and b of (A) (B)

Fig.3 CVs in pH=7.0 buffers at a scan rate of 0.2 V/s for {PAA-PBA/Dex}6-Mb film after {PAA-PBA/Dex}6 film was immersed in 1 mg/mL Mb solutions at pH=5.0 for different time(A) and effect of the immersing time in Mb solution on reduction peak currents of electroactive Mb(Ipc) for {PAA-PBA/Dex}6-Mb film(B)(A) t/h: a. 0; b. 0.5; c. 1.0; d. 2.0; e. 3.0; f. 4.0; g. 5.0; h. 6.0; i. 7.0; j. 9.0; k. 15.0.

Fig.4 Effect of different number of bilayers(n) on the surface concentration of electroactive Mb(Γ*) after {PAA-PBA/Dex}n films were immersed in 1 mg/mL Mb solutions for 9 h

Fig.5 CVs at a scan rate of 0.2 V/s for {PAA-PBA/Dex}6-Mb films in pH=7.0 buffersa. The first cycle; b. after 200 cycles; c. after the films were immersed in pH=7.0 buffers for 7 d.

Fig.6 SEM top views of {PAA-PBA/Dex}6(A), {PAA-PBA/Dex}6/PAA-PBA(B) and {PAA-PBA/Dex}6-Mb films(C) assembled on PG/CS surface

Fig.7 CVs of {PAA-PBA/Dex}6-Mb films at a scan rate of 0.2 V/s in buffers at different pHpH: a. 4.0; b. 6.0; c. 7.0; d. 8.5. Inset: influence of pH of testing solutions on the formal potential(E°') estimated by CV at a scan rate of 0.2 V/s for {PAA-PBA/Dex}6-Mb film.

Fig.8 CVs at before and after different volume of air was injected 0.2 V/s for {PAA-PBA/Dex}6-Mb films in pH=7.0 buffers containing 0.1 mol/L different supporting electrolytesa. NaCl; b. KCl; c. NaBr.

Fig.9 CVs at 0.2 V/s in 10 mL of pH=7.0 buffers for {PAA-PBA/Dex}6(a), {PAA-PBA/Dex}6-Mb films before(b) and {PAA-PBA}6-Mb films(c—j) after different volume of air was injectedV(air)/nL: b. 60; c. 50; d. 10; e. 15; f. 20; g. 30; h. 40; i. 60; j. 90.

Fig.10 CVs at a scan rate of 0.2 V/s for {PAA-PBA/Dex}6 films in buffers(pH=7) containing 100 μmol/L H2O2(a), {PAA-PBA/Dex}6-Mb films(b), {PAA-PBA/Dex}6-Mb films with 20(c), 40(d), 60(e), 80(f), 100(g) and 150 μmol/L(h) H2O2 in pH=7.0 buffers, respectively(A) and dependence of CV Ipc on the concentration of H2O2 for {PAA-PBA/Dex}6-Mb film at 0.2 V/s(B)

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