Immunophilins, a large family of proteins constitute Cyclophilins, FKBPs and Parvulins. Cyclophilin proteins in Triticum aestivum possessing peptidyl prolyl cis trans isomerase(E.C. 35.1.2.8) activity and due to this activity cis trans isomerization of peptidyl prolyl bond observed.cDNA library was used to chosen Cyclophilin A like protein from T. aestivum .Primers used in the study to amplify DNA by PCR and E. coli DH5α used for cloning.The amplified Cyclophilin (CyPA) PCR product was gel purified and concentrations were measured by nanodrop spectrometer as 10ng/?l for Cyclophilin (CyPA)gene . Purification of CYP A protein was done using Ni-NTA chromatography ,the purified CYPA through Ni-NTA column migrated at about 21 kDa respectively in SDS-PAGE and confirmation of recombinant CYPA protein by Western blot analysis observed a 21 kDa band that reacted with anti-His antibodies then Identification of CYPA proteins by mass spectrometry was done. The activity of PPiase was measured by using the coupled assay ,the synthetic peptide N-succinyl-Ala-Xaa-Pro-Phe-p-nitroanilide was used as substrate. In this study we revealed that amplified Cyclophilin (CyPA) PCR product concentrations were measured by nanodrop spectrometer as 10ng/?l for Cyclophilin (CyPA)gene and hyperbolic regression of CYPA protein showed Vm = 0.7094 ? 0.08761 min/?g, Km = 0.7094 ? 0.08761 ?M .