An HPSEC Method for Determining the Cleavage Position of a Protein in Enzymatic Hydrolysis

Abstract

A method for determining the cleavage position of a protein in enzymatic hydrolysis using high performance size‐exclusion chromatography (HPSEC) was established, in terms of an example of bovine serum albumin (BSA) in tryptic hydrolysis. The molecular relative mass distribution of the hydrolysates is characterized by the chromatogram. Comparing the molecular relative mass of theoretical peptides based on the primary structure of BSA with that from experiments, the amino acid sequences of theoretical peptides matching to the peaks in chromatogram are obtained. Thus, the possible cleavage position of BSA in tryptic hydrolysis can be deduced by analyzing the appearing frequency of some amino acid serial.

Keywords:

• Bovine serum albumin
• Trypsin
• Cleavage position
• High performance size‐exclusion chromatography

Acknowledgment

The authors are very grateful for the financial support from the National Natural Science Foundation of China (Grant No. 20276052).