利用IR光谱原位检测生物酶活性的二维相关分析校正

The Correction of 2D Correlation Analysis on the In Situ Enzymatic Assay by IR Spectrum

在利用IR光谱原位监测酶促反应的过程中,反应物与底物红外特征吸收峰会随反应进程发生变化,因此可根据单位时间内特征峰吸收值的变化量来反映酶的催化效率。然而,该方法在实际应用过程中还存在诸多的局限性有待改进。例如,峰位选取不准确和谱带严重重叠无法分辨/分离等都将给酶活力的测试结果带来偏差。二维相关分析与IR光谱的结合使用将在一定程度上改进上述问题,我们将在本文中以具体事例分别阐述。

In the in situ monitoring of enzymatic reaction by using IR spectra the intensities of specific bands will be changed accordingly and,therefore,the time-dependent intensity changes can be used to quantitatively evaluate the enzymatic activity.However,in some cases the bands of substrate and product cannot be separated very well or the specific band position cannot be chosen correctly,leading to inaccuracies in the determination of enzymatic activity.We combined two-dimensional correlation infrared(2D-IR)and infrared difference spectra for their ability to separate overlapped bands.Both the pitfalls and advantages of each method to separate overlapped bands will be critically analyzed in the present study,by using good examples.