New Methods For Fluorescence Analysis Of Enzymes

This paper is consisted of four chapters, including the following aspects: Review on the application and the analysis methods of enzymes; The establishment of a homogeneous method for the determination of proteases using a fluorescent substrate prepared by TMRIC and BSA; The preparation of a acridine orange-cellulose complex and its application in the analysis of cellulase; The preparation of acridine orange-amylum complex and its application in the determination of amylase.Chapter 1 is the preface, introducing practical applications of enzyme in various domains such as medicine, food, industry, environmental protection, etc. In this chapter we come up with the research program of this paper, on the basis of analysis and evaluation of some existing methods for the determination of enzymes. Chapter 2 discusses the homogeneous analysis of proteases by fluorescence anisotropy technique, using rhodamine as the probe. This chapter is consisted of the following parts: (1) Preparation of the TMRITC-BSA; (2) Research on the measurement of Tripsin with TMRITC-BSA as the substrate in alkali environment; (3) Research on the measurement of Bromelain with TMRITC-BSA as the substrate in neutral medium; (4) Research on the measurement of Pepsin with TMRITC-BSA as the substrate in acidic medium; (5) A demo example for illustration purpose. Compared to other previous work, the present method is more independent of pH. Also it widens the application scope of fluorescent anisotropy technology in enzymology analysis area.Chapter 3 presents the main ideas for the measurement of cellulase with acridine orange as the fluorescent probe for the first time. This chapter comprises...