The present invention relates to the technical field of
enzyme activity determination, and discloses a
guanosine deaminase
activity detection method, which comprises: carrying out
centrifugation on a
guanosine deaminase production
fermentation broth, collecting wet thalli, carrying out resuspension by using a PBS
phosphate buffer solution, carrying out
centrifugation again to collect the wet thalli, and recording the
mass of the wet thalli, re-suspending the wet thalli by adopting a
Tris-HCl
buffer solution, adding
lysozyme for
cracking, ultrasonically crushing at low temperature and centrifuging to obtain a crude
enzyme solution, and recording the total volume of the crude
enzyme solution; preparing a
reaction system, adding the crude enzyme solution into the
reaction system to serve as an experimental group, adding a
Tris-HCl
buffer solution with the same amount as the crude enzyme solution into the
reaction system to serve as a blank control group, carrying out oscillation reaction, adding NaOH to terminate the reaction after the reaction is finished, and recording the reaction time; respectively measuring the peak areas of the
xanthosine in the experimental group and the blank control group by using
high performance liquid chromatography, and calculating the content of the
xanthosine by using an external standard method; and finally, calculating to obtain the enzyme activity of each
gram of wet thalli. The detection method is simple, efficient, few in impurities and high in accuracy.