Starch-based double-loaded functional nano particle as well as preparation method and application

Patent metadata

Field	Value
Publication number	US-12083231-B2
Application number	US-202217715970-A
Country	US
Kind code	B2
Filing date	Apr 8, 2022
Priority date	Aug 6, 2021
Publication date	Sep 10, 2024
Grant date	Sep 10, 2024

Abstract

The preparation method of a starch-based double-loaded functional nanoparticle includes: performing restrictive hydrolysis treatment on egg high-density lipoprotein using proteases to obtain the polypeptide; performing self-assembling on a mixed system containing the polypeptide and quercetin under the alkaline condition to form a micelle nanoparticle; performing covalent grafting reaction on a mixed system containing the micelle nanoparticle and anthocyanin under the alkaline condition to form a graft; and electrostatically compounding carboxymethyl dextrin with the graft to obtain the starch-based double-loaded functional nanoparticle. In the preparation method, raw materials derived from natural sources are used, and the self-assembled colloid nanoparticle with good properties can be obtained by adjusting the pH without any organic reagents. The obtained product has a nanoparticle size, has high antioxidant activity and stability against environmental stress, and can be widely applied to the fields of delivery of nutrients, stabilization of biologically active substances and the like.

First claim

What is claimed is:

1. A preparation method of a starch-based double-loaded functional nanoparticle, comprising:
1) performing a restrictive hydrolysis treatment on egg high-density lipoprotein by using proteases, and then isolating and purifying by using dialysis and ethanol precipitation methods to obtain a polypeptide;
2) performing self-assembling on a first mixed system containing the polypeptide and quercetin under a first alkaline condition to form a micelle nanoparticle;
3) performing a covalent grafting reaction on a second mixed system containing the micelle nanoparticle and anthocyanin in a second alkaline environment to form a graft; and
4) Electrostatically compounding carboxymethyl dextrin with the graft to obtain the starch-based double-loaded functional nanoparticle.

2. The preparation method according to claim 1 , wherein in step 1, a temperature of the restrictive hydrolysis treatment is 30-55° C., a time of the restrictive hydrolysis treatment is 0.5-4.5 h, and a pH of a restrictive hydrolysis system is 8.0-9.0, and an addition amount of the protease is 20-100 U/g of the egg high-density lipoprotein.

3. The preparation method according to claim 1 , wherein step 1 comprises:
after the restrictive hydrolysis treatment is completed, dialyzing a hydrolysate resulting from the restrictive hydrolysis treatment at 0-4° C. for more than 48 h, wherein a cutoff molecular weight of an adopted dialysis bag is above 5000 Daltons, then mixing a dialyzed product with ethanol to obtain an ethanol mixture so that a final concentration of ethanol in the ethanol mixture is 10-25%,
centrifuging the ethanol mixture to remove a precipitate, and
subjecting a supernate to freeze drying and then oven-drying at 75-90° C. for 5-20 min to obtain the polypeptide.

4. The preparation method according to claim 1 , wherein the proteases comprise at least one selected from the group consisting of trypsin, alkaline protease, and neutral protease.

5. The preparation method according to claim 1 , wherein in step 1, after the restrictive hydrolysis treatment, a hydrolysis degree of the polypeptide obtained is 6.7%-10%.

6. The preparation method according to claim 1 , wherein in step 1, a molecular weight of the polypeptide is 5-20 kDa.

7. The preparation method according to claim 1 , wherein step 2 comprises:
dispersing the polypeptide and the quercetin into water to form the first mixed system, wherein a concentration of the polypeptide is 10-50 mg/mL and a mass ratio of the quercetin to the polypeptide is 1:100-1:10,
adding NaCl to adjust an ionic strength of the first mixed system to 80-120 mmol/L, then adjusting a pH of the first mixed system to 10-12, and stirring the first mixed system for 30-60 min, so that the polypeptide and the quercetin are subjected to the self-assembling to form the micelle nanoparticle.

8. The preparation method according to claim 1 , wherein step 3 comprises:
mixing a solution of the micelle nanoparticle with a solution of the anthocyanin to form the second mixed system,
adjusting a pH of the second mixed system to 9.0-9.5, and continuously stirring the second mixed system at a speed of 300-600 rpm to undergo the covalent grafting reaction for 20-30 h to form the graft.

9. The preparation method according to claim 8 , wherein in step 3, a mass ratio of the quercetin to the anthocyanin is 1:2-2:1.

10. The preparation method according to claim 1 , wherein step 3 comprises:
after the covalent grafting reaction is completed, adjusting a pH of a reaction mixture to be neutral, and then dialyzing the reaction mixture at 0-4° C. for 20-24 h to obtain the graft, wherein a cutoff molecular weight of an adopted dialysis bag is 3500-4000 Daltons.

11. The preparation method according to claim 1 , wherein step 4 comprises: stirring and mixing a solution of the graft with a solution of the carboxymethyl dextrin to obtain a third mixed system, and meanwhile adding an acidic solution to adjust a pH of the third mixed system to obtain the starch-based double-loaded functional nanoparticle.

12. The preparation method according to claim 1 , wherein a mass ratio of the polypeptide to the carboxymethyl dextrin is 4:1-1:4.

13. The preparation method according to claim 1 , wherein a method for preparing the carboxymethyl dextrin comprises:
alkalizing dextrin with a homogeneous polymerization degree at 35-40° C. for 3-5 h to obtain alkalized dextrin; and
etherifying a mixture containing the alkalized dextrin and monochloroacetic acid in a mass ratio of 1:0.3-1 at 40-45° C. at a pH of 6.5-7.5 for 4-6 h to obtain homogeneous carboxymethyl dextrin with a substitution degree of 0.3-0.7.

14. The preparation method according to claim 13 , wherein a method for preparing the dextrin with the homogeneous polymerization degree Comprises:
a) dispersing rice starch into a phosphate buffer solution with a pH of 5-6 to obtain a rice starch dispersion,
b) heating the rice starch dispersion to obtain a gelatinized rice starch and cooling the gelatinized rice starch to 50-55° C. to obtain a cooled gelatinized rice starch,
c) adding pullulanase to the cooled gelatinized rice starch to perform a debranching treatment for 20-24 h, wherein an addition amount of the pullulanase is 50-100 U/g rice starch,
d) subjecting a product obtained in step c to a water bath treatment, then grading the product obtained after the water bath treatment using an ethanol stepwise precipitation method to obtain the dextrin with the homogeneous polymerization degree of 60-80.

15. A starch-based double-loaded functional nanoparticle, wherein the starch-based double-loaded functional nanoparticle is prepared by the preparation method according to claim 1 .

16. The starch-based double-loaded functional nanoparticle according to claim 15 , wherein the starch-based double-loaded functional nanoparticle is spherical, with a core-shell structure, and possesses a particle size of 189-259 nm and an absolute ζ potential value of 26.1-35.2 mV.

Assignees

• Univ Hefei Technology

Inventors

• Zhang Bao
• Liu Wenjie
• Tao Han
• Li Xiaolong
• Chen Pin
• Wang Rumeng

Classifications

• C12P21/06

  produced by the hydrolysis of a peptide bond, e.g. hydrolysate products (preparing foodstuffs by protein hydrolysis A23J3/00) · CPC title

• C12P19/16

  produced by the action of an alpha-1, 6-glucosidase, e.g. amylose, debranched amylopectin (non-biological hydrolysis of starch C08B30/00) · CPC title

• C12P19/04

  Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds · CPC title

• C08B30/18

  Dextrin {, e.g. yellow canari, white dextrin, amylodextrin or maltodextrin; Methods of depolymerisation, e.g. by irradiation or mechanically} · CPC title

• C07K19/00

  Hybrid peptides {, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes} · CPC title