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Time-dependent nature in peptic hydrolysis of native bovine hemoglobin

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Abstract

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.

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Acknowledgements

The authors thank the financial support from the Natural Science Foundation of China (No. 20306023).

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Authors and Affiliations

  1. Chemical Engineering Research Center, School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, People’s Republic of China

    Rong-Xin Su, Wei Qi & Zhi-Min He

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  1. Rong-Xin Su
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  2. Wei Qi
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  3. Zhi-Min He
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Correspondence to Wei Qi.

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Su, RX., Qi, W. & He, ZM. Time-dependent nature in peptic hydrolysis of native bovine hemoglobin. Eur Food Res Technol 225, 637–647 (2007). https://doi.org/10.1007/s00217-006-0458-y

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  • DOI: https://doi.org/10.1007/s00217-006-0458-y

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