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Antifreeze polypeptide, bionic antifreeze surface related to antifreeze polypeptide, and screening method and application of antifreeze polypeptide

An anti-freezing polypeptide and anti-freezing protein technology, applied in the field of biochemistry, can solve the problems of unsatisfactory anti-icing effect of hydrophobic materials, protein inactivation, complicated preparation methods of super-hydrophobic materials, etc.

Inactive Publication Date: 2014-08-06
INST OF CHEM CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the anti-icing effect of hydrophobic materials is not ideal. The preparation method of superhydrophobic materials is cumbersome, and the combination of antifreeze proteins and polymers requires the modification of reactive groups on proteins, which can easily lead to protein inactivation.

Method used

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  • Antifreeze polypeptide, bionic antifreeze surface related to antifreeze polypeptide, and screening method and application of antifreeze polypeptide
  • Antifreeze polypeptide, bionic antifreeze surface related to antifreeze polypeptide, and screening method and application of antifreeze polypeptide
  • Antifreeze polypeptide, bionic antifreeze surface related to antifreeze polypeptide, and screening method and application of antifreeze polypeptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0105] Screening of antifreeze peptides:

[0106] All kinds of highly active proteins, due to the high thermal hysteresis value of insect antifreeze proteins, and the repeated amino acid sequence Thr-Cys-Thr-X-Ser-X-X-Cys-X-X-Ala-X in insect antifreeze proteins can form a flat The β sheet structure can be well combined with the ice surface. The same repeated amino acid sequence Thr-Cys-Thr-Asp-Ser-Thr-Asn-Cys-Tyr-Lys-Ala-Thr in the antifreeze protein of Tenebrio molitor and A. The negative singlet in the circular dichroism spectrum confirmed the existence of its β-sheet structure. It is found that the polypeptide has a thermal hysteresis value of 0.4° C. in a 1 mg / ml aqueous solution through differential scanning calorimetry measurement. This kind of polypeptide has a high thermal hysteresis value at a lower concentration, and is a preferred antifreeze polypeptide.

Embodiment 2

[0108] Antifreeze polypeptides (proteins) are used in the preparation of antifreeze surfaces:

[0109] Prepare an alkaline buffer solution Bicine (N,N-bishydroxyethylglycine) with a concentration of 10mM / ml, and the pH value is 8.3. Dilute dopamine with alkaline buffer solution Bicine to prepare 2mg / ml dopamine solution. Soak the cleaned silicon wafer in the dopamine solution and soak it in an oxygen environment. The obtained dopamine-modified silicon wafers were rinsed with water, and the silicon wafers were blown dry. Prepare antifreeze polypeptide (protein) solution. The concentration of antifreeze polypeptide (protein) solution is 0.2mg / ml. Soak the silicon chip modified with the polydopamine layer in the antifreeze polypeptide solution. Soak the silicon chip modified with antifreeze polypeptide (protein) in water, soak, repeat this process three times. Blow dry the silicon, or hang to dry. That is, a frost-resistant surface with antifreeze properties is obtained.

Embodiment 3

[0111] Antifreeze surface prepared by antifreeze polypeptide with α-helical structure and its characteristics:

[0112] The antifreeze polypeptide DTA with α-helical structure was bound to the surface of the silicon substrate by the method of Example 2. DTA has a thermal hysteresis value of 0.1°C in a 1 mg / ml solution. The modified silicon substrate was placed in a closed chamber, 0.5uL of water was dropped on the modified surface, and the closed chamber was placed on a cold stage. The surface temperature was raised to 50°C at a rate of 10°C / min, and the temperature was kept at 50°C for 10 minutes to evaporate water droplets in a closed chamber. Decrease from room temperature to minus 40°C at a rate of 5°C / min to condense saturated water vapor on the surface and freeze. Record the freezing temperature of each droplet. The freezing temperature of the water droplets condensed on the surface of the substrate is counted, and the average freezing temperature of the condensed wat...

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Abstract

The invention relates to a method for screening a polypeptide fragment with antifreeze activity. The method comprises a step of screening amino acid sequences of natural antifreeze protein, antifreeze protein or antifreeze glycoprotein or properly improving amino acid sequences of natural antifreeze protein, antifreeze protein or antifreeze glycoprotein. The invention further relates to antifreeze polypeptide, a bionic antifreeze surface related to the antifreeze polypeptide, a coating containing the antifreeze polypeptide and a preparation method and application of antifreeze polypeptide. The antifreeze polypeptide inherits antifreeze characteristics of the antifreeze protein, has a simple structure, is easy to synthesize on large scale and to produce in batches and exerts better effects compared with the antifreeze protein when used for preparation of the antifreeze surface.

Description

technical field [0001] The invention relates to the fields of biochemistry, material science, and surface interface science, and relates to an antifreeze surface prepared by using proteins and polypeptides with thermal hysteresis effects, and a method for preparing a novel bionic antifreeze surface. Background technique [0002] The thermal hysteresis effect is to reduce the freezing point of the solution in a non-colligative manner without changing its melting point, so that the freezing point and melting point of the solution are different, and the difference between the melting point and the freezing point of the solution is called the thermal hysteresis value. [0003] Antifreeze proteins (AFPs) are a class of proteins with thermal hysteresis. According to the adsorption-inhibition mechanism proposed by Raymond and Devries et al., antifreeze proteins can adsorb on the surface of ice crystals, change the growth characteristics of ice crystals, and inhibit the growth of ic...

Claims

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Application Information

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IPC IPC(8): C07K7/08C07K7/06C09K3/18C09D189/00
Inventor 张依帆王健君李开勇宋延林
Owner INST OF CHEM CHINESE ACAD OF SCI
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