Modified porous rice starch-prolamin core-shell structure with probiotic delivery function and preparation method thereof
Patent Information
- Application Number
- CN202510183168.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-02-19
- Publication Date
- 2026-08-21
AI Technical Summary
[0017]本发明的有益效果是：核壳结构的芯材，采用大米淀粉经酶改性、化学改性形成多孔结构，更好地将益生菌吸附、包裹，进一步的玉米醇溶蛋白通过反溶剂沉淀，自组装的方式包裹淀粉颗粒，可以提升淀粉和益生菌在胃肠酸环境的耐受性；本发明操作简单，在实际应用中具有重要作用。
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Figure CN122604072A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of food processing and mainly relates to a modified porous rice starch-prolyzed protein core-shell structure with probiotic delivery function and its preparation method. Background Technology
[0002] Rice starch is a natural polysaccharide extracted from rice and is an important raw material in the food industry and pharmaceutical fields. Due to its good biocompatibility, rice can be used as a drug carrier and excipient. Although untreated rice starch is a valuable natural material, its poor water absorption and solubility, as well as its limitations in functional properties and application areas, make modified starch have significant advantages in practical applications. Appropriate enzymatic hydrolysis can form a porous structure in starch granules, which improves its water absorption and drug release properties. Based on enzymatic modification, chemical modification of porous starch can control the positive charge of starch granules. This functional property can better encapsulate most negatively charged probiotics under natural conditions, effectively improving the utilization value of rice starch.
[0003] Zealysin is a natural biomolecule extracted from corn. Its significant hydrophobicity and slow-release properties in the human body, along with its good bioavailability and gastrointestinal tolerance, make it an attractive plant protein.
[0004] This invention utilizes an antisolvent precipitation method to allow zein to encapsulate modified porous starch loaded with probiotics through self-assembly, providing an important reference for probiotic, drug, or bioactive ingredient delivery systems. Summary of the Invention
[0005] The purpose of this invention is to provide a method for preparing a modified porous rice starch-prolyzed protein core-shell structure with probiotic delivery function.
[0006] The technical problem to be solved by the present invention is achieved through the following technical solution: a modified porous rice starch-prolyzed protein core-shell structure with probiotic delivery function and its preparation method, characterized by comprising the following steps:
[0007] (1) Step 1: Preparation of porous rice starch: Prepare a rice starch suspension with a concentration of 1 g / mL using a 0.1 mol / L sodium acetate buffer solution with a pH of 5.6. Stir in a water bath at 50 ℃ for 20 min to form a starch suspension reaction system. Add 6% of a mixed enzyme solution of α-amylase and saccharifying enzyme to the suspension. Shake and react at 50 ℃ for 24 h. Add 4% NaOH to stop the enzyme reaction. Centrifuge at 5000 r / min for 5 min, discard the supernatant, repeat washing three times, and then dry, grind, and sieve to obtain porous rice starch.
[0008] (2) Step 2, Modification of porous rice starch: Take the porous rice starch prepared in step 1, prepare a starch suspension with a concentration of 10% using deionized water, add a certain amount of 0.1 g / mL polyethyleneimine, stir in a water bath at 300 r / min for 6 h at 50 ℃, add a certain amount of 1% glutaraldehyde, continue stirring in a water bath for 10 h, stop the reaction, centrifuge at 5000 r / min for 5 min, collect the precipitate and wash it repeatedly with deionized water 3 times, dry, grind and sieve to obtain modified porous rice starch.
[0009] (3) Step 3, preparation of probiotics: Inoculate probiotics into MRS liquid medium, and after static culture at 40℃ for 24 h, transfer them to new MRS liquid medium at an inoculation rate of 1% and culture them at high density in a fermenter for 16-24 h. Then, centrifuge the fermentation broth at 4℃ and collect the bacterial cells. After washing with sterile physiological saline, suspend them in sterile physiological saline and prepare a bacterial solution with a certain bacterial cell concentration.
[0010] (4) Step 4, core material preparation: Take 100 mL of the probiotic suspension obtained in step 3, add 2 g of the modified porous rice starch obtained in step 2, shake and react for 2 h, centrifuge at 5000 r / min for 5 min, discard the supernatant, collect the precipitate, wash it 3 times, and then suspend it in 100 mL of sterile distilled water.
[0011] (5) Step 5: Preparation of wall material: Prepare a 2% concentration alcohol-soluble protein solution with a solvent mass fraction of 70%. Heat the alcohol-soluble protein solution at 95°C for 30 min. After heating, cool it to room temperature in an ice bath.
[0012] (6) Step 6, preparation of core-shell structure: Take the starch aqueous solution prepared in step 4 and add it dropwise to the alcohol-soluble protein solution at a certain ratio. The dropwise addition process is at 0.5 mL / min, and the mixture is continuously stirred at 150 r / min until the addition is completed. The mass fraction of ethanol in the final mixed solution is ≤50%. The mixed solution is allowed to stand for 2 h to precipitate, centrifuged at 5000 r / min for 5 min, the precipitate is collected, and washed three times with sterile deionized water to obtain modified porous rice starch-alcohol-soluble protein core-shell structure microparticles.
[0013] In step one, the ratio of α-amylase to saccharifying enzyme is 1:1, 1:3, 1:5, 1:7, 1:9, or 1:11.
[0014] In step three, the probiotics can be Lactobacillus plantarum, Streptococcus thermophilus, Bifidobacterium, Lactobacillus bulgaricus, and Rhamnosus bacillus.
[0015] In step three, the bacterial cell concentration is 10. 6CFU / ml, 10 7 CFU / ml, 10 8 CFU / ml, 10 9 CFU / ml, 10 10 CFU / ml.
[0016] In step six, the mass fraction of ethanol in the final mixed solution is 48%, 45%, 42%, 39%, and 36%.
[0017] The beneficial effects of this invention are: the core material with a core-shell structure is made of rice starch that has been enzymatically and chemically modified to form a porous structure, which better adsorbs and encapsulates probiotics; furthermore, the zein protein is precipitated by antisolvent and self-assembles to encapsulate starch particles, which can improve the tolerance of starch and probiotics in the acidic environment of the gastrointestinal tract; this invention is simple to operate and plays an important role in practical applications. Attached Figure Description
[0018] Figure 1 This is a flowchart illustrating the preparation process of core-shell structured particles. Detailed Implementation
[0019] The following detailed embodiments will provide an in-depth analysis of the implementation details of this application. It should be understood that any equivalent modifications made by those skilled in the art under the inspiration of this invention should be considered within the scope of protection of this invention. The illustrated embodiments are only a part of the numerous implementations of this invention, and not all of them.
[0020] Example 1
[0021] (1) Step 1: Preparation of porous rice starch: Prepare a rice starch suspension with a concentration of 1 g / mL using a 0.1 mol / L sodium acetate buffer solution with a pH of 5.6. Stir in a water bath at 50 ℃ for 20 min to form a starch suspension reaction system. Add 6% of a mixed enzyme solution of α-amylase and saccharifying enzyme to the suspension. Shake and react at 50 ℃ for 24 h. Add 4% NaOH to stop the enzyme reaction. Centrifuge at 5000 r / min for 5 min, discard the supernatant, repeat washing three times, and then dry, grind, and sieve to obtain porous rice starch.
[0022] (2) Step 2, Modification of porous rice starch: Take the porous rice starch prepared in step 1, prepare a starch suspension with a concentration of 10% using deionized water, add a certain amount of 0.1 g / mL polyethyleneimine, stir in a water bath at 300 r / min for 6 h at 50 ℃, add a certain amount of 1% glutaraldehyde, continue stirring in a water bath for 10 h, stop the reaction, centrifuge at 5000 r / min for 5 min, collect the precipitate and wash it repeatedly with deionized water 3 times, dry, grind and sieve to obtain modified porous rice starch.
[0023] (3) Step 3, preparation of probiotics: Inoculate probiotics into MRS liquid medium, and after static culture at 40℃ for 24 h, transfer them to new MRS liquid medium at an inoculation rate of 1% and culture them at high density in a fermenter for 16-24 h. Then, centrifuge the fermentation broth at 4℃ and collect the bacterial cells. After washing with sterile physiological saline, suspend them in sterile physiological saline and prepare a bacterial solution with a certain bacterial cell concentration.
[0024] (4) Step 4, core material preparation: Take 100 mL of the probiotic suspension obtained in step 3, add 2 g of the modified porous rice starch obtained in step 2, shake and react for 2 h, centrifuge at 5000 r / min for 5 min, discard the supernatant, collect the precipitate, wash it 3 times, and then suspend it in 100 mL of sterile distilled water.
[0025] (5) Step 5: Preparation of wall material: Prepare a 2% concentration alcohol-soluble protein solution with a solvent mass fraction of 70%. Heat the alcohol-soluble protein solution at 95°C for 30 min. After heating, cool it to room temperature in an ice bath.
[0026] (6) Step 6, preparation of core-shell structure: Take the starch aqueous solution prepared in step 4 and add it dropwise to the alcohol-soluble protein solution at a certain ratio. The dropwise addition process is at 0.5 mL / min, and the mixture is continuously stirred at 150 r / min until the addition is completed. The mass fraction of ethanol in the final mixed solution is 42%. The mixed solution is allowed to stand for 2 h to precipitate, centrifuged at 5000 r / min for 5 min, the precipitate is collected, and washed three times with deionized water to obtain modified porous rice starch-alcohol-soluble protein core-shell structure microparticles.
[0027] Example 2
[0028] (1) Step 1: Preparation of porous rice starch: Prepare a rice starch suspension with a concentration of 1 g / mL using a 0.1 mol / L sodium acetate buffer solution with a pH of 5.6. Stir in a water bath at 50 ℃ for 20 min to form a starch suspension reaction system. Add 6% of a mixed enzyme solution of α-amylase and saccharifying enzyme to the suspension. Shake and react at 50 ℃ for 24 h. Add 4% NaOH to stop the enzyme reaction. Centrifuge at 5000 r / min for 5 min, discard the supernatant, repeat washing three times, and then dry, grind, and sieve to obtain porous rice starch.
[0029] (2) Step 2, Modification of porous rice starch: Take the porous rice starch prepared in step 1, prepare a starch suspension with a concentration of 10% using deionized water, add a certain amount of 0.1 g / mL polyethyleneimine, stir in a water bath at 300 r / min for 6 h at 50 ℃, add a certain amount of 1% glutaraldehyde, continue stirring in a water bath for 10 h, stop the reaction, centrifuge at 5000 r / min for 5 min, collect the precipitate and wash it repeatedly with deionized water 3 times, dry, grind and sieve to obtain modified porous rice starch.
[0030] (3) Step 3, preparation of probiotics: Inoculate probiotics into MRS liquid medium, and after static culture at 40℃ for 24 h, transfer them to new MRS liquid medium at an inoculation rate of 1% and culture them at high density in a fermenter for 16-24 h. Then, centrifuge the fermentation broth at 4℃ and collect the bacterial cells. After washing with sterile physiological saline, suspend them in sterile physiological saline and prepare a bacterial solution with a certain bacterial cell concentration.
[0031] (4) Step 4, core material preparation: Take 100 mL of the probiotic suspension obtained in step 3, add 2 g of the modified porous rice starch obtained in step 2, shake and react for 2 h, centrifuge at 5000 r / min for 5 min, discard the supernatant, collect the precipitate, wash it 3 times, and then suspend it in 100 mL of sterile distilled water.
[0032] (5) Step 5: Preparation of wall material: Prepare a 2% concentration alcohol-soluble protein solution with a solvent mass fraction of 70%. Heat the alcohol-soluble protein solution at 95°C for 30 min. After heating, cool it to room temperature in an ice bath.
[0033] (6) Step 6, preparation of core-shell structure: Take the starch aqueous solution prepared in step 4 and add it dropwise to the alcohol-soluble protein solution at a certain ratio. The dropwise addition process is at 0.5 mL / min, and the mixture is continuously stirred at 150 r / min until the addition is completed. The mass fraction of ethanol in the final mixed solution is 45%. The mixed solution is allowed to stand and precipitate for 2 h, centrifuged at 5000 r / min for 5 min, the precipitate is collected, and washed three times with deionized water to obtain modified porous rice starch-alcohol-soluble protein core-shell structure microparticles.
[0034] Example 3
[0035] (1) Step 1: Preparation of porous rice starch: Prepare a rice starch suspension with a concentration of 1 g / mL using a 0.1 mol / L sodium acetate buffer solution with a pH of 5.6. Stir in a water bath at 50 ℃ for 20 min to form a starch suspension reaction system. Add 6% of a mixed enzyme solution of α-amylase and saccharifying enzyme to the suspension. Shake and react at 50 ℃ for 24 h. Add 4% NaOH to stop the enzyme reaction. Centrifuge at 5000 r / min for 5 min, discard the supernatant, repeat washing three times, and then dry, grind, and sieve to obtain porous rice starch.
[0036] (2) Step 2, Modification of porous rice starch: Take the porous rice starch prepared in step 1, prepare a starch suspension with a concentration of 10% using deionized water, add a certain amount of 0.1 g / mL polyethyleneimine, stir in a water bath at 300 r / min for 6 h at 50 ℃, add a certain amount of 1% glutaraldehyde, continue stirring in a water bath for 10 h, stop the reaction, centrifuge at 5000 r / min for 5 min, collect the precipitate and wash it repeatedly with deionized water 3 times, dry, grind and sieve to obtain modified porous rice starch.
[0037] (3) Step 3, preparation of probiotics: Inoculate probiotics into MRS liquid medium, and after static culture at 40℃ for 24 h, transfer them to new MRS liquid medium at an inoculation rate of 1% and culture them at high density in a fermenter for 16-24 h. Then, centrifuge the fermentation broth at 4℃ and collect the bacterial cells. After washing with sterile physiological saline, suspend them in sterile physiological saline and prepare a bacterial solution with a certain bacterial cell concentration.
[0038] (4) Step 4, core material preparation: Take 100 mL of the probiotic suspension obtained in step 3, add 2 g of the modified porous rice starch obtained in step 2, shake and react for 2 h, centrifuge at 5000 r / min for 5 min, discard the supernatant, collect the precipitate, wash it 3 times, and then suspend it in 100 mL of sterile distilled water.
[0039] (5) Step 5: Preparation of wall material: Prepare a 2% concentration alcohol-soluble protein solution with a solvent mass fraction of 70%. Heat the alcohol-soluble protein solution at 95°C for 30 min. After heating, cool it to room temperature in an ice bath.
[0040] Step 6: Preparation of core-shell structure: Take the starch aqueous solution prepared in Step 4 and add it dropwise to the alcohol-soluble protein solution at a certain ratio. The dropwise addition is carried out at 0.5 mL / min, and the mixture is stirred continuously at 150 r / min until the addition is completed. The mass fraction of ethanol in the final mixed solution is 48%. The mixed solution is allowed to stand for 2 h to precipitate, centrifuged at 5000 r / min for 5 min, the precipitate is collected, and washed three times with deionized water to obtain modified porous rice starch-alcohol-soluble protein core-shell structured microparticles.
Claims
1. A modified porous rice starch-prolyzed protein core-shell structure with probiotic delivery function and its preparation method, characterized in that, Includes the following steps: Step 1: Preparation of porous rice starch: Prepare a rice starch suspension with a concentration of 1 g / mL using a 0.1 mol / L sodium acetate buffer solution at pH 5.
6. Stir in a water bath at 50 ℃ for 20 min to form a starch suspension reaction system. Add 6% of a mixed enzyme solution of α-amylase and saccharifying enzyme to the suspension. Shake and react at 50 ℃ for 24 h. Add 4% NaOH to stop the enzyme reaction. Centrifuge at 5000 r / min for 5 min, discard the supernatant, repeat washing three times, and then dry, grind, and sieve to obtain porous rice starch. Step 2: Modification of porous rice starch: Take the porous rice starch prepared in Step 1, prepare a starch suspension with a concentration of 10% using deionized water, add a certain amount of 0.1 g / mL polyethyleneimine, stir in a water bath at 300 r / min for 6 h at 50 ℃, add a certain amount of 1% glutaraldehyde, continue stirring in a water bath for 10 h, stop the reaction, centrifuge at 5000 r / min for 5 min, collect the precipitate, wash it repeatedly with deionized water 3 times, dry, grind, and sieve to obtain modified porous rice starch; Step 3, Probiotic Preparation: Inoculate probiotics into MRS liquid medium and incubate at 40℃ for 24 h. Then, transfer 1% of the inoculum into new MRS liquid medium and culture at high density in a fermenter for 16-24 h. Centrifuge the fermentation broth at 4℃ to collect the cells. Wash with sterile physiological saline and suspend in sterile physiological saline to prepare a bacterial solution with a certain cell concentration. Step 4, Core Material Preparation: Take 100 mL of the probiotic suspension obtained in Step 3, add 2 g of the modified porous rice starch obtained in Step 2, shake and react for 2 h, centrifuge at 5000 r / min for 5 min, discard the supernatant, collect the precipitate, wash it 3 times, and then suspend it in 100 mL of sterile distilled water. Step 5, wall material preparation: Prepare a 2% concentration alcohol-soluble protein solution with a solvent mass fraction of 70%. Heat the alcohol-soluble protein solution at 95℃ for 30 min. After heating, cool it to room temperature in an ice bath. Step 6: Preparation of core-shell structure: Take the starch aqueous solution prepared in Step 4 and add it dropwise to the alcohol-soluble protein solution at a certain ratio. The dropwise addition is carried out at 0.5 mL / min, and the mixture is stirred continuously at 150 r / min until the addition is completed. The mass fraction of ethanol in the final mixed solution is ≤50%. The mixed solution is allowed to stand for precipitation for 2 h, centrifuged at 5000 r / min for 5 min, the precipitate is collected, and washed three times with sterile deionized water to obtain modified porous rice starch-alcohol-soluble protein core-shell structured microparticles.
2. The modified porous rice starch-prolyzed protein core-shell structure with probiotic delivery function according to claim 1 and its preparation method, characterized in that: In step one, the ratio of α-amylase to saccharifying enzyme is 1:1, 1:3, 1:5, 1:7, 1:9, or 1:
11. Preferably, the ratio of α-amylase to saccharifying enzyme is 1:
5.
3. The modified porous rice starch-prolyzed protein core-shell structure with probiotic delivery function according to claim 1 and its preparation method, characterized in that: In step three, the probiotics can be Lactobacillus plantarum, Streptococcus thermophilus, Bifidobacterium, Lactobacillus bulgaricus, and Rhamnosus bacillus.
4. The modified porous rice starch-prolyzed protein core-shell structure with probiotic delivery function according to claim 1 and its preparation method, characterized in that: In step three, the bacterial cell concentration is 10. 6 CFU / ml, 10 7 CFU / ml, 10 8 CFU / ml, 10 9 CFU / ml, 10 10 CFU / ml, preferably, the bacterial concentration is 10. 8 CFU / ml.
5. The modified porous rice starch-prolyzed protein core-shell structure with probiotic delivery function according to claim 1 and its preparation method, characterized in that: In step six, the mass fraction of ethanol in the final mixed solution is 48%, 45%, 42%, 39%, or 36%, preferably 42%.