关键词: 生物分子, 电化学现场, 拉曼光谱学

Abstract: Electron?transfer reaction is known to be one of the key reactions for generating biological functions. Mechanism revelation at a molecular level of such kind reactions is to be very helpful for us to understand life essence. In fact, surface enhanced Raman scattering (SERS) is one of the most powerful tools for the study on metal?electrolyte and metal?vacuum interfaces since 1970's. Moreover, Raman spectroscopic study in enzymology has provided attractive results during last twenty?five years. For the study of electron?transfer reaction mechanism of some oxidoreductases and SERS of some other biological macromolecules, an electrochemical in situ Raman spectroscopic technique was established in author's lab and some research works have been done on it in the past two years. A brief review of these works is given in this paper. The electrochemical in situ Raman spectroscopic measurements were carried out using a Super LABRAM Raman spectrometer (Dilor, France) coupled with a CHI604A Electrochemical Analyzer (CH Instr., USA). A Teflon spectroelectrochemical cell with a quartz plate window was designed for the in situ measurements. The working electrode was pretreated with oxidation?reduction cycles for each measurement. The electrolyte solutions were purged with nitrogen prior to all measurements, and all the measurements were carried out under the nitrogen atmosphere. Copper, zinc superoxide dismutase (SOD) is an important oxidoreductase for organism metabolism. The established spectroelectrochemical technique was first used to characterize the cyclic voltammetric process of SOD at L?cysteine modified gold electrode as well as the process of electrochemical modification of L?cysteine molecules on a gold electrode. The obtained Raman spectra reveal that the L?cysteine modified gold electrode improves effectively the reversibility of electron?transfer reactions of SOD. Besides L?cysteine molecules, it was interesting that adenine was also an effective electron?transfer promoter for SOD at gold electrode. A strong peak at 355 cm -1 can be observed in the Raman spectrum of adenine molecules adsorbed on gold electrode. It was inferred that the peak maybe related to the chemical interaction between adenine molecules adsorbed and gold electrode surface. As shown in Fig.1, for the mixture of SOD and adenine at gold electrode under a polarization potential 55 mV (vs. SCE), both the characteristic Raman lines of SOD and adenine molecules appeared. Therefore it was reasonable to conclude that SOD and adenine molecules should be co?adsorbed on gold electrode surface under such a potential, which is slightly lower than the reduction peak potential of SOD on adenine?modified gold electrode. Moreover, two new peaks appeared remarkably at 445 cm -1 and 610 cm -1 are likely to be related to the active site of SOD. It suggests that the co?adsorption mechanism of SOD and adenine molecules on the gold electrode surface results in effective approaching of the active site of SOD to the electrode surface.