European Food Research and Technology

, Volume 225, Issue 5, pp 637–647

Time-dependent nature in peptic hydrolysis of native bovine hemoglobin

Original Paper

DOI: 10.1007/s00217-006-0458-y

Cite this article as:
Su, RX., Qi, W. & He, ZM. Eur Food Res Technol (2007) 225: 637. doi:10.1007/s00217-006-0458-y


Native bovine hemoglobin was digested with pepsin at pH 4.5 in a batched stirred tank reactor. The resulting peptides in the time-course samples were separated and identified by RP-HPLC coupled with ESI-MS/MS. The secondary structure of bovine hemoglobin was not significantly changed after acid-treatment at pH 4.5 based on far-UV circular dichroism spectra. The α-helix contents of peptic hydrolysates decreased gradually with time according to ‘one-by-one’ mechanism. MS/MS analysis enabled unambiguous identification of 31 and 15 peptides released respectively from α-chain and β-chain, which resulted in their sequence coverage of 100 and 76%. The discrimination of peptic susceptibility between different protein areas was compared in terms of the time-dependent release of peptides. At first, peptic cleavage sites were concentrated around N- and C-terminal regions of α-chain and β-chain. Later, pepsin hydrolyzed the middle part of α-chain from N- to C-terminal, while little enzymatic cleavage occurred in the center region of β-chain due mainly to their high hydrophilic nature. The release kinetics of peptides was discussed in relation to the hydrophobicity of amino acid residues of polypeptide chains.


HemoglobinPepsinProtein hydrolysisEnzymatic kineticsTandem mass spectrometryCircular dichroism

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  1. 1.Chemical Engineering Research CenterSchool of Chemical Engineering and Technology, Tianjin UniversityTianjinPeople’s Republic of China