Identification and Characterization of Bovine (Bos taurus) Pancreatic Bile Salt Activated Lipase for Potential Halal Alternative.

Abstract

Introduction: Bile salt-activated lipase (BSAL) is one of the pancreatic lipases that plays a critical role in the digestion and absorption of dietary fats. Objective: This study aimed to purify bile salt-activated lipase (BSAL) from bovine (Bos taurus) pancreatic tissue. Methods: Bovine pancreas was freshly collected from Abattoir Complex, Department of Veterinary Services, Shah Alam, Malaysia. The fats were removed by trimming grossly visible fat, and extraction was performed using organic solvents. The BSAL was further purified by anion exchange chromatography and sent for protein identification by liquid chromatography-mass spectrometry (LC-MS/MS). Results: Successful purification of bovine BSAL was visualized as a single protein band on sodium dodecyl sulfate-polyacrylamide (SDS) gel, which LC-MS/MS confirmed as a bovine BSAL (Accession number – P30122) with a molecular mass of 65.12 kDa and calc pI of 5.57. Peptide identification based on the MS spectrum found 200 predictive peptides, of which ten sequences with bovine BSAL peptide characteristics. The selected predictive peptide sequences have a molecular mass of 1104.60 - 3378.94 Da with Qvality q-value greater than 0.01 and XCorr Sequest HT value ranging from 2.6 to 6.8. The specific lipolytic enzyme activity of bovine BSAL was comparable with the positive control, as measured using lipase assay. Conclusion and Recommendations: In conclusion, the results of this study indicate the effectiveness of bovine BSAL purification by anion exchange chromatography from fresh pancreatic tissue and have the potential for further Halal pharmaceuticals and medical applications.