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Compositional analysis of proteins following hydrolysis by immobilized proteases.

F C Church, H E Swaisgood, G L Catignani

    Journal of Applied Biochemistry
    |August 1, 1984
    PubMed
    Summary

    Combining immobilized enzymes with acid hydrolysis improves protein quality assessment. This method achieves total protein hydrolysis efficiently, offering a more accurate chemical score than acid hydrolysis alone.

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    Area of Science:

    • Biochemistry
    • Analytical Chemistry
    • Protein Chemistry

    Background:

    • Accurate protein quality assessment is crucial in nutrition and food science.
    • Conventional acid hydrolysis for amino acid analysis can degrade certain amino acids.
    • Enzymatic hydrolysis offers a milder alternative but may be incomplete.

    Purpose of the Study:

    • To evaluate the efficacy of immobilized proteases for total protein hydrolysis.
    • To compare the accuracy of combined enzymatic and acid hydrolysis with acid hydrolysis alone for protein quality assessment.
    • To determine the time efficiency of immobilized enzyme hydrolysis.

    Main Methods:

    • Immobilization of various proteases (Pronase, Proteinase K, carboxypeptidases A/B, aminopeptidase M, etc.) onto controlled-pore glass beads.

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  • Enzymatic hydrolysis of protein substrates (beta-lactoglobulin, insulin) using immobilized enzymes.
  • Combined enzymatic and acid hydrolysis for protein quality assessment.
  • Amino acid analysis of hydrolysates.

    • Main Results:

    • Immobilized enzymes achieved essentially total hydrolysis of protein substrates.
    • Amino acid analysis of enzymic hydrolysates yielded 92% of theoretical values.
    • Combined hydrolysis provided a more accurate chemical score than acid hydrolysis alone.
    • Hydrolysis time (18-24 h) was comparable to conventional acid hydrolysis.

    Conclusions:

    • A combination of immobilized enzymes provides efficient and comprehensive protein hydrolysis.
    • This method enhances the accuracy of protein quality assessment compared to acid hydrolysis alone.
    • Immobilized enzyme technology offers a viable alternative for protein analysis.