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

An anti-freeze polypeptide, low temperature technology, applied in the field of anti-freeze polypeptide, can solve the problems of loss of quality, damage to cell and tissue structure, etc., and achieve the effect of high anti-freeze activity

Active Publication Date: 2015-04-15
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

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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-thaw damage and a preparation method thereof. Fish-derived collagen is used as a raw material, and alkaline protease is used for enzymolysis, separation, purification, and freeze-drying to obtain low-temperature freeze-thaw protection for bacteria. Damaged antifreeze polypeptide, the complete amino acid sequence 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. The invention breaks through the existing research ideas and methods of antifreeze protein (polypeptide) at home and abroad, overcomes the limitation of the quantity of antifreeze protein purified from natural organisms and the safety of transgenic antifreeze protein in food application by the international FDA organization Concerns, obtain high-efficiency antifreeze polypeptides based on food sources, and lay a theoretical foundation for the development of antifreeze polypeptides based on food sources and their wide application in food and medicine.

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 Patents(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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