红外(IR)光谱近年来逐渐展示了它在生物医学领域中的应用,它不但能很好地用于体外酶催化活力分析和酶抑制剂的筛选,尤其可对活细胞进行原位酶活力测定和抑制剂筛选。IR光谱技术可用于生物酶活力的直接检的基本原理是基于单位时间内,反应物或者产物在IR光谱中特征吸收峰的峰位或强度的变化量来反映生物酶的催化效率。本文综述了利用IR光谱技术,对体外部分酶活力值进行分析和测定、以及对酶抑制剂的抑制效应进行评估和筛选;进而将该技术扩展至以活体细胞作为检测模型、开展细胞内生物酶活力值的测定及其抑制剂的筛选。同时,我们也对新近提出的在细胞原位测定酶活力的IR方法及研究理念作了概述,生理条件下开展对生物酶活性无标记直接检测提供了新方法,并对该技术领域存在的问题及今后的发展趋势作出了展望。
A real-time infrared(IR)spectroscopy measurement is an effective means to obtain enzymatic information either in vitro or in living cells,as it can provide direct,continue test of biomacromolecule reactions.The principles of measurements are performed basing on the fact that the absorption bands in spectra of reactants and products are usually separated to each other and changed independently with time.Therefore,it is possible to measure the enzymatic efficiency at any reaction time according to the changes of characteristic band,from either reactant or product.That is,IR spectroscopy can be used to obtain intracellular structural information during the cells metabolic processes,as which can provide detailed and reliable scientific evidences.In this paper,we summarized the new developments of IR spectra in the in vitro enzymatic assay for several representative enzymes,as well the screening of enzyme inhibitors,which was further extended to the identical aspect by using living cells as detection model.Such important enzymatic examination closes to the physiological conditions without labeling,supplying structural information of the related biomolecules.The developing trends of IR spectra are discussed and the perspectives of it in the research area are also provided in this review.