Keywords

Hydrolysis, Gelatin, Antifreeze Peptide, Isolation

Abstract

Alkaline protease is used to hydrolyze food-derived gelatin to prepare antifreeze peptides. By controlling the enzymatic hydrolysis conditions, a gelatin peptide enzymatic hydrolyzate with antifreeze activity is obtained, and then subjected to chromatographic separation, mass spectrometry identification and full amino acid sequence determination. The results show that when the ratio of enzyme to substrate is 1:15, the enzymatic hydrolysis time is 30 minutes, and the enzymatic hydrolysis temperature is 45¡ãC, the antifreeze activity of the gelatin polypeptide enzymatic hydrolyzate obtained is the highest; the gelatin polypeptide liquid prepared by enzymatic hydrolysis undergoes cold-heat The inhibition of ice crystal growth and antifreeze activity after circulation (cold-heat-stage) showed specific rules, indicating that the control and optimization of enzymatic hydrolysis conditions is the key to achieving the antifreeze activity of gelatin peptide liquid. Through Sephadex G-50, SP Sephadex C-25, C18-RPLC chromatographic separation and MALDI-TOF mass spectrometry identification, it was determined that a specific polypeptide with a molecular mass of 2107D showed significant antifreeze activity. Its full amino acid sequence was determined by Edman degradation method to be GERGFPGERGSPGAQGLQGPR

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Original research done by Zhou Yanfu, Wang Shaoyun, Zhao Lina, Rao Pingfan

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