Keywords

Antifreeze Peptides, Isolation, Cryoprotection

Abstract

Use hot water extraction method to extract shark skin gelatin, and control enzymatic hydrolysis conditions to prepare natural antifreeze peptides. Using bacterial low-temperature protection activity as an indicator, screen the types of proteolytic enzymes and enzymatic hydrolysis time, and use Sephadex G-50 gel filtration chromatography, SP-Sephadex C-25 strong cation exchange chromatography and other separation methods were used to separate the highly active components of bacterial low-temperature protection. The results showed that the most suitable proteolytic enzyme for enzymatic hydrolysis of shark skin gelatin to obtain highly active antifreeze polypeptides is acid protease. Hydrolysis temperature 50 ¡æ, pH 3.0, enzyme/substrate 3 000 U¡¤g -1, substrate concentration 3%, enzymatic hydrolysis time 1 h; passed Sephadex G-50 gel filtration chromatography, SP-Sephadex C-25 strong The cationic Pª­2 component was obtained by separation methods such as cation exchange chromatography. After bacterial low-temperature protection testing, when the concentration was 500 ¦Ìg¡¤mL-1, the survival rate of E. coli reached 80.8%. The isolated antifreeze polypeptide has high It has antifreeze activity and is expected to be used in food, pharmaceutical and other industries.

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Original research done by Zhao Jun, Wang Shaoyun, Li Xiaokun

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