Antifreeze polypeptide for protecting bacteria from low temperature freeze-thawing damage and preparation method thereof

An antifreeze polypeptide and low temperature technology, applied in the field of antifreeze polypeptide, can solve problems such as loss of quality, damage to cells and tissue structures, and achieve high antifreeze activity

Active Publication Date: 2013-12-25
FUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Repeated rise and fall of temperature causes ice crystals to grow continuously, freeze-thaw and recrystallize, which seriously damages the cell and tissue structure and loses the quality of the product.

Method used

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  • Antifreeze polypeptide for protecting bacteria from low temperature freeze-thawing damage and preparation method thereof
  • Antifreeze polypeptide for protecting bacteria from low temperature freeze-thawing damage and preparation method thereof
  • Antifreeze polypeptide for protecting bacteria from low temperature freeze-thawing damage and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0026] Weigh 1.65 g of fish skin collagen and dissolve it in 6 ml of Mili-Q water, then adjust its pH to 9.0 with 2 mol / L NaOH. First heat the solution to 45°C in a water bath, then add the corresponding amount of enzyme according to the enzyme-substrate ratio of 1:20, and the enzymatic hydrolysis time is 30 minutes. Then inactivate the enzyme in a boiling water bath for 10 minutes, then centrifuge at 14000 rpm for 10 minutes after cooling, and collect the supernatant for later use.

[0027] The supernatant was separated by Sephadex G-50 gel chromatography (length 100cm, diameter 2.6cm), the eluent was deionized water, the flow rate was 2mL / min, and the elution peak was measured at 225nm. Elution peak of antifreeze activity.

[0028] The elution peak with the best antifreeze activity separated by Sephadex G-50 gel chromatography is separated in the next step, separated by Sulfopropyl-Sepadex C-25 cation exchange chromatography (length 55cm, diameter 2.0cm), and eluted The so...

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Abstract

The invention discloses an antifreeze polypeptide for protecting bacteria from low temperature freeze-thawing damage and a preparation method thereof. The preparation method is characterized by using fish-derived collagens as raw materials, adopting alkaline protease to carry out enzymolysis on the fish-derived collagens and carrying out separation and purification and freeze drying, thus obtaining the antifreeze polypeptide for protecting bacteria from low temperature freeze-thawing damage. The amino acid complete sequence of the antifreeze polypeptide is Gly-Gln-Arg-Gly-Gly-Arg-Gly-Leu-Pro-Gly-Glu-Arg-Gly-Arg-Val-Gly-Pro-Ser-Gly-Pro-Ala-Gly-Ala-Arg-Gly-Ala-Asp-Gly. By adopting the antifreeze polypeptide and the preparation method, the research ideas and methods of antifreeze proteins (polypeptides) existing at home and abroad are broken through, limitation of the quantity of the antifreeze proteins purified form natural organisms and considerations of international FDA (food and drug administration) for security of transgenic antifreeze proteins in food application are overcome, an efficient antifreeze polypeptide based on food sources is obtained, and a theoretical foundation is laid for developing the antifreeze polypeptide based on food sources and exploring extensive application of the antifreeze polypeptide to food and medicines.

Description

technical field [0001] The invention relates to an antifreeze polypeptide, more specifically to an antifreeze polypeptide prepared by hydrolyzing fish-source collagen with alkaline protease, and belongs to the field of biotechnology. Background technique [0002] The problem that food and pharmaceutical products repeatedly suffer from ice crystal growth and recrystallization due to fluctuations in ambient temperature during cryogenic storage and transportation has attracted increasing attention. The repeated rise and fall of temperature causes ice crystals to grow continuously, freeze-thaw and recrystallize, which seriously damages the cell and tissue structure and loses the quality of the product. Scientists around the world are facing serious challenges: how to control the growth and recrystallization of ice crystals, and realize the control of ice crystal growth on the low-temperature cold chain is the key to restricting the quality of many food and pharmaceutical product...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/46A01N1/02C12P21/06C07K1/20C07K1/18
Inventor 汪少芸邵彪赵立娜饶平凡
Owner FUZHOU UNIV
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