Enzymes are being routinely used in clinical laboratory for diagnosis of various diseases. It is imperative to have reference values of clinically important enzymes for accurate interpretation of laboratory data as well as for their correlation with relevant diseases. Though numerous studies have been carried out on blood biochemical profile of various domestic animal species, the Assam local sheep remain untouched in this aspect. Present study reports the physiological values of major diagnostic enzymes viz. SGPT, SGOT, LDH, ALP, GGT and Amylase as 20.6±1.68 U/L, 170.8±3.96 U/L, 534.6±9.91 U/L, 163.8±41.82 U/L, 154.2±65.38 and 92.2±11.47 U/L, respectively. The data generated may be of use for interpretation of laboratory data to confirm a disease along with the signs and symptoms.