In Vitro Activity of CYP2J Recombinant Protease From Bactrian Camel

The purpose of this experiment was to study the in vitro activity of recombinant protease CYP2J of Bactrian camel. The activity of CYP2J enzyme was determined by fluorescence method according to the change of fluorescence peak after the conversion of ethoxycoumarin to hydroxycoumarin in NADPH generation system and ethoxycoumarin deethylase reaction system. Then, the affinity of the recombinant protease with its specific substrates arachidonic acid (AA) and astemizole was detected by localised surface plasmon resonance (LSPR) technique, and its activity was further determined. The results showed that the standard curve is y=17 533x+190.73 and R²=0.998; The fluorescence detection result of the repeated experiments was 0.1350±0.0251 nM/min/mg, and the significant value between the repeated experiments was greater than 0.05, indicating that the difference was not statistically significant. The affinity of the recombinant protease for arachidonic acid was 3.53×10^-5 M and that for astemizole was 4.07×10^-5 M, respectively. Therefore, the Bactrian camel CYP2J recombinant protease has sufficient stable activities in vitro and can meet the basic requirements of further research.