Preparation of Glucan-based Shell-core Structure Carrier Material and Its Application thereof

Abstract

The present invention discloses a glucan-based shell-core structure carrier material and preparation and application thereof, and belongs to the technical field of modern food processing. Spherical hyperbranched water-soluble amylum grains are used as the raw material, and an enzymatic grafting and chain extending process is adopted for treatment to modify the surfaces of water-soluble glucan molecules into a firm shell structure with densely cumulated crystal structures, and form the glucan-based carrying material with the shell-core structure of which an inner core cavity has an amorphous state and an outer shell layer has a crystalline state. The adopted spherical hyperbranched water-soluble amylum grains have wide sources of raw materials and are not limited by producing areas and seasons; the preparation has simple and convenient steps, easy operation, controllable reaction conditions, relatively low cost and basically no pollution to the environment; and the prepared product can effectively protect, deliver and release functional nutritional components, can be applied to multiple fields of food, medicine, chemicals for daily use and the like, and has great market prospects and broad economic benefits.

Claims

1. A glucan-based shell-core structure carrier material, wherein the glucan-based shell-core structure carrier material is obtained by performing grafting and chain extending on glucosyl groups on outer surfaces of spherical hyperbranched water-soluble amylum grains by -1,4 glycosidic bonds by using glycosyltransferase.

2. The glucan-based shell-core structure carrier material of claim 1, wherein the molecular weight of the water-soluble amylum grains is 10.sup.7-10.sup.8 g/mol, the proportion of -1,6 glycosidic bonds is 7%-10%, and an average particle size is 30-100 nm.

3. The glucan-based shell-core structure carrier material of claim 1, wherein the spherical hyperbranched water-soluble amylum grains are from one or more of natural plant spherical hyperbranched starch granules, oyster glycogen in animals and biotechnology-synthesized high-molecular spherical polysaccharides.

4. The glucan-based shell-core structure carrier material of claim 1, wherein the spherical hyperbranched water-soluble amylum grains are from sweet-type soluble corn glucan.

5. The glucan-based shell-core structure carrier material of claim 1, wherein the glucan-based shell-core structure carrier material is prepared by preparing a solution from the water-soluble amylum grains first, then performing a reaction in a system containing donor molecules for providing glucose molecules and the glycosyltransferase, and performing enzyme deactivation, centrifugation, drying and precipitation after the reaction to obtain the glucan-based shell-core structure carrier material.

6. The glucan-based shell-core structure carrier material of claim 5, wherein the mass ratio of the donor molecules for providing glucose molecules to the water-soluble amylum grains is (1.5:1) to (5:1).

7. The glucan-based shell-core structure carrier material of claim 1, wherein donor molecule for providing glucose molecules is glucose-1-phosphate.

8. The glucan-based shell-core structure carrier material of claim 7, wherein the glucose-1-phosphate comprises a sodium salt or a potassium salt.

9. The glucan-based shell-core structure carrier material of claim 1, wherein a functional component is added during grafting and chain extending.

10. The glucan-based shell-core structure carrier material of claim 1, wherein the glycosyltransferase comprises glycogen phosphorylase and -glucose phosphorylase.

11. The glucan-based shell-core structure carrier material of claim 1, wherein during preparation, sweet-type soluble corn glucan is used as a main raw material, and grafting and chain extending are performed on spherical hyperbranched corn glucan by using the glycosyltransferase to form the glucan-based shell-core structure carrier material.

12. The glucan-based shell-core structure carrier material of claim 1, wherein the preparation comprises the following specific processing steps: (1) weighing 1 g of water-soluble amylum grains and dissolving the amylum grains in a buffer solution to prepare a uniform solution with the mass concentration of 0.5-3.0%; (2) according to the proportions of 1.5-5 g of glucose-1-phosphate to 10-180 U of glycosyl transferase per 1 g of water-soluble amylum grains, adding the glucose-1-phosphate and the glycosyltransferase, performing uniform stirring, and performing a thermostatic reaction at temperature of 35-40 C. and pH value of 6.5-7.5 for 3-24 h; (3) performing enzyme deactivation by heating and centrifugation treatment, and performing vacuum drying on the obtained precipitate to obtain the glucan-based shell-core structure carrier material.

13. A complex embedded with a functional component, wherein the complex is prepared by adding the functional component to a reaction system in a formation process of the glucan-based shell-core structure carrier material of claim 1.

14. A biological carrying material, containing the glucan-based shell-core structure carrier material of claim 1.

15. A method to use the glucan-based shell-core structure carrier material of claim 1 to carry a functional active substance in the aspect of carrying.

Description

BRIEF DESCRIPTION OF FIGURES

[0033] FIG. 1 is schematic diagrams of a carrying material with a shell-core structure and a complex of the carrying material with the shell-core structure and nutritional factors;

[0034] FIG. 2 is X-ray diffraction pattern results of original amylum grains and the carrying material with the shell-core structure.

DETAILED DESCRIPTION

[0035] To better realize the present invention, biological stability is characterized by determining the oxidation rate POV (peroxide value) by applying a potassium thiocyanate POV determining method. POV is calculated through the formula below:

[00001]$P.Math..Math.O.Math..Math.V\left(\frac{m.Math..Math.\mathrm{equiv}}{\mathrm{kg}}\right)=\frac{\left(c-c.Math..Math.0\right)}{\left(m55.841\right)},$

[0036] c and c.sub.0 are the mass of iron in a test sample and a blank sample; m is the mass of CLA; 2 is a conversion factor; and 55.84 is the relative atomic mass of iron. The degree of oxidation of pure nutritional factors serves as control, and the maximum values of the amount of peroxide (POV) in the control and the material are calculated and compared. POV is the amount of peroxide in a first stage product obtained after fat oxidation. Because it cannot be excluded that a small amount of peroxide continues to decompose into small molecular substances under an oxidizing environment, the stability is characterized by:

Stability(100100 g CLA maximum value of the amount of peroxide generated by oxidation)/100*100%.

[0037] Cell experiment: Intestinal cell experiment is performed on a carrying complex. 100 L of a carrying material-nutritional factor complex dissolved solution is added to a cell culture solution, 2 mM of a hydrogen peroxide solution is added for irritating cells for 2 h, and the cells are continued to be cultured for 4 h. Cell activity is detected by an MTT method.

Example 1

[0038] 1 g of spherical hyperbranched water-soluble amylum grains (sweet-type soluble corn glucan) are weighted, and the amylum grains are dissolved in a Tris-HCl buffer solution (50 mmol/L, pH7.0) for preparing a uniform solution with the mass concentration of 0.5%. 1.5 g of glucose-1-phosphate and 40 U of glycosyltransferase is continually added, uniform stirring is performed, and a thermostatic reaction at the temperature of 40 C. and the pH value of 7.0 is performed for 12 h. Enzyme deactivation by heating and centrifugation treatment are performed, and vacuum drying is performed on the obtained precipitate to obtain the glucan-based shell-core structure carrier material.

[0039] As shown in FIG. 1, a represents the spherical hyperbranched water-soluble amylum grains; b represents the early stage of grafting and chain extending when the amylum grains are modified by a biotechnology, i.e., glucosyl groups are sequentially connected to non-reducible terminals of the spherical starch granules by -1,4 glycosidic bonds by the glycosyltransferase; c represents the later stage of grafting and chain extending after the amylum grains are modified by the biotechnology, i.e., linear chain structures formed by grafting are wound and crosslinked on the outer surfaces of the spherical amylum grains, double spiral structures are formed in partial positions, and further a shell-core structure of which an inner core cavity has an amorphous state and an outer shell layer has a crystalline state is formed by accumulation and gathering; d represents the complex of the carrying material and nutritional factors obtained by a vine winding method, i.e., due to interior hydrophobicity and exterior hydrophilicity, linear chain single spiral structures generated by grafting and chain extending can include the nutritional factors to form the complex of the carrying material and the nutritional factors by hydrophobic interaction.

[0040] FIG. 2 is X-ray diffraction pattern results of original amylum grains and the carrying material with the shell-core structure. The results show that an amorphous form is changed into a certain crystal structure.

TABLE-US-00001 TABLE 1 Properties of Carrying Material with Shell-core Structure Sample Crystallinity (% 3%) Crystal size (nm) Original amylum 0 0 Carrying material with 23.54 5.084 shell-core structure

Example 2

[0041] 1 g of spherical hyperbranched water-soluble amylum grains (sweet-type soluble corn glucan) are weighted, and the amylum grains are dissolved in a Tris-HCl buffer solution (50 mmol/L, pH7.0) for preparing a uniform solution with the mass concentration of 1.0%. 2.5 g of glucose-1-phosphate and 60 U of glycosyltransferase are continually added, uniform stirring is performed, and a thermostatic reaction at the temperature of 40 C. and the pH value of 7.0 is performed for 18 h. Enzyme deactivation by heating and centrifugation treatment are performed, and vacuum drying is performed on the obtained precipitate to obtain the glucan-based shell-core structure carrier material.

Example 3

[0042] 1 g of spherical hyperbranched water-soluble amylum grains (sweet-type soluble corn glucan) are weighted, and the amylum grains are dissolved in a Tris-HCl buffer solution (50 mmol/L, pH7.0) for preparing a uniform solution with the mass concentration of 1.5%. 5.0 g of glucose-1-phosphate and 100 U of glycosyltransferase are continually added, uniform stirring is performed, and a thermostatic reaction at the temperature of 40 C. and the pH value of 7.0 is performed for 24 h. Enzyme deactivation by heating and centrifugation treatment are performed, and vacuum drying is performed on the obtained precipitate to obtain the glucan-based shell-core structure carrier material.

Example 4: Application of Glucan-Based Shell-Core Structure Carrier Material

[0043] The glucan-based shell-core structure carrier material prepared in Examples 1-3 is applied to carrying of a functional active substance conjugated linoleic acid. A specific test method is as follows:

[0044] Nutritional factors are added during the grafting and chain extending reaction of amylum grains, i.e., 1 g of spherical hyperbranched amylum grains are dissolved in a buffer solution, 5.0 g of glucose-1-phosphate, 1000 of enzymes and 10 mg of nutritional factors conjugated linoleic acid dissolved in a small amount of ethanol are continually added, mixing and uniform stirring are performed, and a thermostatic reaction at the temperature of 40 C. and the pH value of 7.0 is performed for 24 h. Shell-core structure is formed after grafting and chain extending are finished, and because the structure contains single spiral cavities with hydrophobic effects, the structure can include nutrients to further form a complex. 15% sodium chloride solution can be added to accelerate the generation of products, and the carrying complex is obtained by performing centrifugation, washing with 50% alcohol and drying treatment.

TABLE-US-00002 TABLE 2 Conjugated Linoleic Acid-Carried Glucan-based Shell-core Structure Carrier Material Biological Uplift ratio of stability biological stability Embodiment 1 91.4% 33.2% Embodiment 2 94.6% 36.7% Embodiment 3 97.3% 39.1% Control 1 58.2% / Control 2 88.7% 30.5% Control 3 89.5% 31.3%

[0045] the control 1 is a blank control group, namely the nutritional factor conjugated linoleic acid.

[0046] In an implementation method of the control 2: amylose is dissolved in a dimethyl sulfoxide solution at 90 C., cooling is preformed to 30 C., the dimethyl sulfoxide solution containing the amylose is mixed with dimethyl sulfoxide containing conjugated linoleic acid with the same temperature, and single spiral-nutritional factor inclusion is finished; 20 times of deionized water and a 15% sodium chloride solution with the same temperature by volume are added to accelerate generation of products, and centrifugation, washing with 50% alcohol and drying treatment are performed to obtain a carrying material-nutritional factor complex.

[0047] In an implementation method of the control 3: 20 mg of maltose and 200 mg of glucose-1-phosphate are dissolved in 100 nM of a citric acid buffer solution (pH 7.0) containing 5 nM of adenosine monophosphate and 20 U of D-enzyme, 1 mg of phosphorylase is added, a reaction is performed at the temperature of 30 C. for 2 h, centrifugation is performed on reactant liquor, supernatant liquor is treated at 100 C. for 5 min, denaturase protein is removed by centrifugation, 50 U of glucoamylase is added to the supernatant liquor, and the precipitate is ring structure glucan; the obtained ring structure glucan is dissolved in a dimethyl sulfoxide solution at 90 C., cooling is performed to 30 C., the dimethyl sulfoxide solution containing the ring structure glucan is mixed with dimethyl sulfoxide containing conjugated linoleic acid with the same temperature, and single spiral-nutritional factor inclusion is finished; 20 times of deionized water and a 15% sodium chloride solution with the same temperature by volume are added to accelerate generation of products, and centrifugation, washing with 50% alcohol and drying treatment are performed to obtain the carrying material-nutritional factor complex.

Example 5: Application of Glucan-Based Shell-Core Structure Carrier Material

[0048] The glucan-based shell-core structure carrier material prepared in Example 3 is applied to carrying of a functional active substance coenzyme Q10.

[0049] A specific test method is as follows:

[0050] Nutritional factors are added during the grafting and chain extending reaction of amylum grains, i.e., 1 g of amylum grains are dissolved in a buffer solution, 10.0 g of glucose-1-phosphate, 100 U of enzyme and 10 mg of nutritional factors coenzyme Q10 dissolved in a small amount of ethanol are continually added, mixing and uniform stirring are performed, and a thermostatic reaction at the temperature of 40 C. and the pH value of 7.0 is performed for 24 h. A shell-core structure is formed after grafting and chain extending are finished, and because the structure contains single spiral cavities with hydrophobic effects, the structure can include nutrients to further form a complex. A 15% sodium chloride solution can be added to accelerate the generation of products, and the carrying complex is obtained by performing centrifugation, washing with 50% alcohol and drying treatment.

TABLE-US-00003 TABLE 3 Intestinal Cell Experiment Cell Hydrogen peroxide solution activity % 44 2.0 Coenzyme Q10 Carrying material coenzyme Q10 complex (1 g/ml) (1 g/ml) 92 3.4 59 2.7 Coenzyme Q10 Carrying material coenzyme Q10 complex (10 g/ml) (10 g/ml) 97 2.6 82 3.1

Example 6: Optimized Research on Conditions in Material Preparation Process

[0051] (1) Spherical hyperbranched water-soluble amylum grains in 1 g of sweet-type soluble corn glucan are dissolved in a Tris-HCl buffer solution (50 mmol/L, pH7.0) for preparing a uniform solution with the mass concentration of 5.0%, and other conditions referred to the example 3 are unchanged to prepare a carrier material.

[0052] (2) Refer to the example 3, the addition amount of the glucose-1-phosphate is changed from 1.5 g to 10 g, and other conditions are unchanged to prepare a carrier material.

[0053] (3) Refer to the example 3, the addition amount of the glycosyltransferase is changed from 40 U to 250 U, and other conditions are unchanged to prepare a carrier material.

[0054] By referring to the example 4 and respectively applying the carrier materials obtained by the above three methods to carrying of a functional active substance conjugated linoleic acid, it is discovered that the biological stability of the 3 kinds of carrying complexes is relatively low, and does not exceed 75%, wherein the carrier material in the method (3) has no obvious improvement as compared with the control 1 (blank load).

[0055] The specific embodiments described herein are merely illustration of the spirit of the present invention and some of the experiments. A person skilled in the art can make various modifications or complements to the specific embodiments described or replace them in a similar manner, without departing from the spirit of the present invention or beyond the scope of defined in the appended claims.