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CN-121970743-A - Form-adjustable antifreeze polypeptide nanostructure and preparation method thereof

CN121970743ACN 121970743 ACN121970743 ACN 121970743ACN-121970743-A

Abstract

The invention discloses a morphological adjustable antifreeze polypeptide nanostructure and a preparation method thereof, comprising the following steps of S1, preparing a solution containing polypeptide molecules, S2, quick-freezing the solution in the step S1 into ice to enrich the polypeptide molecules at the interface between ice crystals, S3, regulating and controlling the temperature of the ice obtained in the step S2 to age the ice crystals, and further, enabling the polypeptide molecules to self-assemble in a limited space at the interface to form the nanostructure with obvious antifreeze activity. The problem that the self-assembled morphology of the traditional polypeptide is uncontrollable is successfully solved by utilizing the ice water interface concentration effect and combining the solvent annealing technology, and the precise adjustability of morphology (such as spherical shape and rod shape) on the nanometer scale is realized. The prepared antifreeze agent can effectively inhibit the growth and recrystallization of ice crystals under extremely low concentration, and provides a brand new technical path for developing new generation of green low-cost low-temperature protection materials.

Inventors

  • DING ZHONGXIANG
  • LIU HONGLIN
  • CHEN HAIJIE

Assignees

  • 安徽理工大学

Dates

Publication Date
20260505
Application Date
20251230

Claims (10)

  1. 1. The preparation method of the morphological adjustable antifreeze polypeptide nanostructure is characterized by comprising the following steps of: S1, preparing a solution containing polypeptide molecules; s2, quick-freezing the solution in the step S1 into ice, so that polypeptide molecules are enriched at the interface between ice crystals; S3, regulating and controlling the temperature of the ice obtained in the step S2 to enable the ice crystal to be cured, and further enabling the polypeptide molecules to self-assemble in a limited space at the interface to form a nano structure with obvious freezing resistance activity.
  2. 2. The method according to claim 1, wherein the polypeptide molecule comprises at least one amino acid selected from the group consisting of threonine, alanine, phenylalanine, tyrosine, proline, and glycine, and wherein the N-terminus or the C-terminus of the polypeptide molecule is modified with an aromatic ring group.
  3. 3. The preparation method of claim 1, wherein the solution further contains at least one of salt ions, water and an organic solvent, wherein the salt ions are cations of at least one of sodium ions and magnesium ions, and anions of at least one of sulfate ions, acetate ions, nitrate ions and halogen ions, and the organic solvent is at least one of dimethyl sulfoxide, methanol, ethanol, acetic acid and acetone.
  4. 4. The method of claim 1, wherein the number of amino acids in the polypeptide molecule is not less than two.
  5. 5. The method of claim 1, wherein the concentration of polypeptide molecules in the solution is 0.01-10 mg/mL.
  6. 6. The method of claim 1, wherein in step S2, the freezing temperature is at least 40 ℃ lower than the melting point of the solution.
  7. 7. The method according to claim 1, wherein in step S3, the temperature of the ice obtained in step S2 is controlled, specifically, by annealing or natural melting.
  8. 8. The method of claim 7, wherein the annealing treatment satisfies at least one of the following conditions: (1) The annealing temperature is at least 1 ℃ lower than the melting point of the salt solution; (2) The temperature is raised to the annealing temperature at the rate of 0.5-15 ℃ per minute; (3) The holding time at the annealing temperature is 1 to 30 minutes.
  9. 9. A morphologically adjustable antifreeze polypeptide nanostructure produced by the method of any one of claims 1 to 8, wherein the morphology of said polypeptide nanostructure comprises nanofibers, nanoplatelets or nanowires, the dimensions of which are dynamically adjustable as a function of said annealing temperature, annealing time and/or salt ion type.
  10. 10. The morphologically tunable antifreeze polypeptide nanostructure of claim 9, wherein the concentration effective to inhibit ice crystal growth and recrystallization is from 0.1 to 1.0 mg/mL.

Description

Form-adjustable antifreeze polypeptide nanostructure and preparation method thereof Technical Field The invention relates to the technical field of anti-freezing bionic materials and low-temperature preservation, in particular to a morphological adjustable anti-freezing polypeptide nano structure and a preparation method thereof. Background At present, antifreeze proteins (Antifreeze Proteins, AFPs) and polypeptides have become research hotspots in the field of low-temperature preservation due to their biocompatibility and efficient ice crystal inhibition. Traditional preparation methods rely mainly on chemical synthesis or biological fermentation to directly obtain polypeptide molecules, or to immobilize polypeptides on surfaces by physical adsorption to achieve functionalization. However, the existing polypeptide antifreeze agent still faces a significant technical bottleneck in application: The structure is uncontrollable, polypeptide molecules usually exist in a random coil or random self-assembly form, and the nanoscale morphology (such as spheres, rods and the like) of the polypeptide molecules is difficult to accurately regulate and control through simple process parameters, so that the dispersion stability of the polypeptide molecules at low temperature is poor. Performance limitations although some studies have shown that polypeptides of specific sequences can alter ice crystal morphology and size (e.g., inhibit round ice crystal growth), complex screening or high cost is often required to achieve effective ice recrystallization inhibition at low concentrations. Disclosure of Invention In view of the above, the invention provides a method for preparing a morphological adjustable antifreeze polypeptide nanostructure, which solves the problems in the prior art, successfully solves the problem that the self-assembled morphology of the traditional polypeptide is uncontrollable by utilizing the ice water interface concentration effect in combination with a solvent annealing technology, and realizes the precise adjustability of the morphology (such as sphere and rod shape) on the nanoscale. The prepared antifreeze agent can effectively inhibit the growth and recrystallization of ice crystals under extremely low concentration, and provides a brand new technical path for developing new generation of green low-cost low-temperature protection materials. In order to achieve the above purpose, the present invention provides the following technical solutions: In a first aspect, the invention discloses a method for preparing a morphological adjustable antifreeze polypeptide nanostructure, which comprises the following steps: S1, preparing a solution containing polypeptide molecules; s2, quick-freezing the solution in the step S1 into ice, so that polypeptide molecules are enriched at the interface between ice crystals; S3, regulating and controlling the temperature of the ice obtained in the step S2 to enable the ice crystal to be cured, and further enabling the polypeptide molecules to self-assemble in a limited space at the interface to form a nano structure with obvious freezing resistance activity. Further, the polypeptide molecule contains at least one amino acid of threonine, alanine, phenylalanine, tyrosine, proline and glycine, and the N end or the C end of the polypeptide molecule is modified by an aromatic ring group. The solution further comprises at least one of salt ions, water and an organic solvent, wherein the salt ions are cations of at least one of sodium ions and magnesium ions, and anions of at least one of sulfate ions, acetate ions, nitrate ions and halogen ions, and the organic solvent is at least one of dimethyl sulfoxide, methanol, ethanol, acetic acid and acetone. Further, the number of amino acids in the polypeptide molecule is not less than two. Further, the concentration of polypeptide molecules in the solution is 0.01-10 mg/mL. In a further embodiment, in step S2, the freezing temperature is at least 40 ℃ below the melting point of the solution. In the step S3, the temperature of the ice obtained in the step S2 is regulated and controlled, and the ice is subjected to annealing treatment or natural melting. Polypeptide assembly is driven by controlling the temperature of ice to initiate ice crystal maturation. The temperature regulation and control can be realized in two main modes, namely, annealing treatment, namely, heating ice and keeping the ice at a certain temperature lower than the melting point of the ice for constant-temperature curing, and direct melting, namely, directly heating the ice to be above the melting point, and curing and assembling are completed in the melting process. The annealing treatment can be used for accurately regulating and controlling the morphology of the nano structure. Further, the annealing treatment meets at least one of the following conditions: (1) The annealing temperature is at least 1 ℃ lower than the melting point of the salt solution; (2)