CN116508994A - Probiotics RG-I pectin microcapsule and preparation method thereof - Google Patents

Probiotics RG-I pectin microcapsule and preparation method thereof Download PDF

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CN116508994A
CN116508994A CN202310500051.XA CN202310500051A CN116508994A CN 116508994 A CN116508994 A CN 116508994A CN 202310500051 A CN202310500051 A CN 202310500051A CN 116508994 A CN116508994 A CN 116508994A
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pectin
microcapsule
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叶兴乾
陈品
陈健乐
程焕
陈士国
潘海波
田金虎
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Zhejiang University ZJU
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/015—Inorganic compounds
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/20—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents
    • A23L29/206—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents of vegetable origin
    • A23L29/231—Pectin; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
    • A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
    • A23L3/3463—Organic compounds; Microorganisms; Enzymes
    • A23L3/3562—Sugars; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
    • A23P10/00—Shaping or working of foodstuffs characterised by the products
    • A23P10/30—Encapsulation of particles, e.g. foodstuff additives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Polymers & Plastics (AREA)
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  • Dispersion Chemistry (AREA)
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  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The invention discloses a probiotic RG-I pectin microcapsule and a preparation method thereof, and relates to the technical field of biology. The preparation method comprises the following steps: (1) Adding probiotic bacterial suspension and calcium carbonate solution into RG-I pectin water solution, and uniformly mixing to obtain mixed solution; (2) Adding an oil phase containing an emulsifier into the mixed solution for emulsification reaction, then adding an oil phase containing glacial acetic acid, continuing to react, finally adding a sodium acetate solution, standing, separating to obtain a water phase, sieving, and washing to obtain the RG-I pectin microcapsule. The probiotics RG-I pectin microcapsule provided by the invention can enhance the tolerance of probiotics to severe environment, reduce the loss of probiotics in the gastrointestinal tract digestion process and prolong the shelf life of the probiotics.

Description

Probiotics RG-I pectin microcapsule and preparation method thereof
Technical Field
The invention relates to the technical field of biology, in particular to a probiotic RG-I pectin microcapsule and a preparation method thereof.
Background
Probiotics are important food functional factors, play an important role in the physiological processes of digestion, intestinal immune development, intestinal colonization by pathogens and the like, and have been widely studied. However, probiotics are affected by many factors (such as temperature, humidity, nutrients, acidity, etc.) during processing and storage, so that the activity of the probiotics is reduced and the physiological functions of the probiotics cannot be exerted. And when the probiotics are ingested by human body, the special environment of human digestive tract (gastric acid, bile salt, digestive tract peristalsis, etc.) can also affect the activity of the probiotics. Thus, the implementation of effective embedding techniques for probiotics, thereby enhancing their viability in harsh environments, is of great importance for their commercialization. The microcapsule is a capsule in which a small amount of active ingredient is encapsulated into a micron-sized capsule, and the active ingredient is protected from the surrounding environment by creating a functional barrier between the active ingredient and the environment, avoiding potentially harmful chemical and physical reactions. The method has the advantages of good protectiveness, size controllability, simplicity in operation and the like, and becomes a research hot spot for improving the viability of probiotics.
Although the microcapsules may reach a specific target area for release without being adversely affected by environmental factors. However, this also places a limit on the choice of microcapsule wall material. Rhamnogalacturonan I (RG-I) pectin is one of the three domains that make up pectin, and because of steric hindrance between its backbone and the rhamnose carboxyl of the neutral side chain and the cross-linker, both cross-links are limited, resulting in the inability to form a gel. Thus, RG-I pectin is considered unsuitable as a wall material for microcapsules. Although it was found that RG-I pectin can combine with divalent metal ions to form a gel when its degree of esterification is small, it is still necessary to further study whether it can form microcapsules to protect probiotics.
Disclosure of Invention
The invention aims to provide a probiotic RG-I pectin microcapsule and a preparation method thereof, so as to solve the problems in the prior art.
In order to achieve the above object, the present invention provides the following solutions:
the invention provides a preparation method of a probiotic RG-I pectin microcapsule, which comprises the following steps:
(1) Adding probiotic bacterial suspension and calcium carbonate solution into RG-I pectin water solution, and uniformly mixing to obtain mixed solution;
(2) Adding an oil phase containing an emulsifier into the mixed solution for emulsification reaction, then adding an oil phase containing glacial acetic acid, continuing to react, then adding a sodium acetate solution, standing, separating to obtain a water phase, sieving, and washing to obtain the RG-I pectin microcapsule.
Further, in step (1), the concentration of RG-I pectin in the aqueous RG-I pectin solution is 0.8-1.8wt%.
Further, in step (1), the mass ratio of RG-I pectin in the RG-I pectin aqueous solution to calcium carbonate in the calcium carbonate solution is 1:1-5.
Further, in step (2), the emulsifier comprises tween 80.
Further, the volume fraction of the emulsifier in the oil phase containing the emulsifier is 0.5-2.5%.
Further, in the step (2), after the oil phase containing glacial acetic acid is added, the volume ratio of water to oil in the reaction system is 1:1-5.
The invention also provides the probiotic RG-I pectin microcapsule prepared by the preparation method.
The invention also provides a preparation method of the probiotics RG-I pectin microcapsule containing the biological film, which comprises the step of carrying out stimulated culture on the probiotics RG-I pectin microcapsule to enable the probiotics embedded in the probiotics RG-I pectin microcapsule to form the biological film, so as to obtain the probiotics RG-I pectin microcapsule containing the biological film.
Further, the culture medium for the stimulation culture is added with calcium carbonate, and the addition amount is 10-100mM.
The invention also provides the probiotics RG-I pectin microcapsule containing the biological film, which is prepared by the preparation method.
The invention discloses the following technical effects:
according to the invention, the RG-I pectin microcapsule is prepared by using an endogenous emulsification method, and the probiotics are embedded, so that the tolerance of the probiotics to severe environments is enhanced, the loss of the probiotics in the gastrointestinal digestion process is reduced, and the quality guarantee period of the probiotics is prolonged.
The invention uses Ca 2+ As a cross-linking agent of RG-I pectin, the RG-I pectin is successfully formed into microcapsules, and the application of the RG-I pectin is widened.
While microcapsules can provide a physical barrier to probiotics, the protection of the probiotics by the microcapsules is still insufficient. The adoption of the multi-layer embedding mode can cause the volume increase of the microcapsules, thereby affecting the ingested taste and causing the cost increase. According to the method, the probiotics can grow to generate the biological film, so that the tolerance of the probiotics to the environment is improved, and therefore, after the RG-I pectin is formed into the microcapsule to embed the probiotics by using an endogenous emulsification method, the probiotics form the biological film in the microcapsule by stimulating culture.
Detailed Description
Various exemplary embodiments of the invention will now be described in detail, which should not be considered as limiting the invention, but rather as more detailed descriptions of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In addition, for numerical ranges in this disclosure, it is understood that each intermediate value between the upper and lower limits of the ranges is also specifically disclosed. Every smaller range between any stated value or stated range, and any other stated value or intermediate value within the stated range, is also encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the invention described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from consideration of the specification of the present invention. The specification and examples of the present invention are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are intended to be inclusive and mean an inclusion, but not limited to.
Lactobacillus plantarum (Lactobacillus plantarum) and Lactobacillus rhamnosus (Lactobacillus rhamnosus) used in the examples below were purchased from the university of Cantonese institute of microorganisms.
RG-I pectin is extracted from acidic wastewater and alkaline wastewater discharged from canned citrus factories. The specific method comprises the following steps: the acidic wastewater is filtered through a 400-mesh filter bag. Then, pH was adjusted to 3 (up to 4) with NaOH and precipitated at a volume fraction of 95% ethanol in a volume ratio of 1:1 for 2h to give PA pectin. And the alkaline wastewater is firstly regulated to 6 (up to 7) by using 2MHCl, after natural precipitation, the precipitation is washed by ethanol with the volume fraction of 95%, and the washed precipitation is placed in an oven and dried for 24 hours at 55 ℃ to obtain PB pectin. Finally, mixing PA pectin and PB pectin according to a mass ratio of 1:1 to obtain RG-I pectin.
Commercial pectins were purchased from Shanghai biosciences, inc.
Example 1
(1) Activating strains and preparing a strain suspension: glycerol culture solutions of lactobacillus plantarum and lactobacillus rhamnosus stored at-80 ℃ are respectively inoculated into a sterile MRS liquid culture medium, and are cultured for 24 hours at 37 ℃ to activate the bacteria twice under the same conditions. Inoculating activated strain into MRS liquid culture medium according to 2% (v/v), culturing at 37deg.C, collecting bacterial liquid at late stage of logarithmic growth, centrifuging at 8000r/min for 10min, washing with physiological saline (9 g/L NaCl) for 2 times, and re-suspending with physiological saline to obtain about 10 9 CFU/mL of a bacterial suspension of two probiotics.
(2) Preparation of lactobacillus plantarum/lactobacillus rhamnosus microcapsules: RG-I pectin is dissolved in 20mL of sterile water to a final concentration of 1.5wt% (0.8-1.8 wt% can achieve the same effect), and then mixed with 10mL of lactobacillus plantarum/lactobacillus rhamnosus bacterial suspension, and then added with 0.5g of CaCO 3 (CaCO 3 The mass ratio of the RG-I pectin to the RG-I pectin is controlled at 1:1-5), and the mixture is stirred uniformly to obtain a mixed solution.
Then, 40mL of soybean oil containing Tween 80 (Tween 80 volume fraction is 0.1%, tween 80 volume fraction is required to be controlled to be 0.5-2.5%) is added into the mixed solution, and emulsification is carried out at a speed of 300r/min (200-800 r/min can achieve the same effect) for 15 minutes. Subsequently, 20mL of soybean oil containing glacial acetic acid (volume fraction of glacial acetic acid 0.5%) was added and stirred for 30min. Wherein, after the soybean oil containing glacial acetic acid is added, the volume ratio of water to oil in the reaction system is controlled to be 1:1-5.
Finally, 60mL of sodium acetate solution (volume fraction of sodium acetate is 0.5%) was added, and the mixture was allowed to stand for 60min. After all the formed microcapsules are settled to the bottom of the sodium acetate solution, separating to obtain a water phase, sieving with a 200-mesh sieve (up to 400 meshes), collecting the obtained microcapsules, washing twice, removing residual oil phase and surface bacteria to obtain lactobacillus plantarum/lactobacillus rhamnosus microcapsules, and then storing in a refrigerator at 4 ℃.
(3) 1mL of plant is takenLactobacillus/Lactobacillus rhamnosus microcapsules containing 50mM CaCO 3 Culturing for 12 hours at 37 ℃ to obtain the lactobacillus plantarum/lactobacillus rhamnosus microcapsule containing the biological membrane. Wherein CaCO 3 The addition amount in MRS liquid culture medium is controlled to be 10-100mM.
(4) Determination of microcapsule embedding rate: respectively taking 1mL of the lactobacillus plantarum/lactobacillus rhamnosus microcapsule prepared in the step (2) and the lactobacillus plantarum/lactobacillus rhamnosus microcapsule containing the biological membrane prepared in the step (3), adding the lactobacillus plantarum/lactobacillus rhamnosus microcapsule into 9mL of sterile PBS (pH 7.5) solution, homogenizing the mixture at a speed of 10000r/min for 30s, and then slightly shaking the mixture in a rotary stirrer for 10min to release bacteria in the microcapsules. The total number of colonies was measured by the plate coating method. The formula of encapsulation efficiency (EE%) is EE (%) = (total number of colonies released in microcapsules (Log CFU/mL)/total number of colonies of bacterial suspensions of two probiotics for microcapsule preparation (Log CFU/mL)) ×100%.
The embedding rate results show that the embedding rate of lactobacillus plantarum in the microcapsule is 65.08 percent and the embedding rate of lactobacillus rhamnosus is 65.95 percent.
Example 2
The biological membrane-containing lactobacillus plantarum/lactobacillus rhamnosus microcapsule prepared in example 1 is subjected to gastrointestinal digestion tolerance detection, and the specific steps are as follows:
into a test tube containing 8mL of simulated gastric fluid, 2mL of the Lactobacillus plantarum suspension, lactobacillus rhamnosus suspension, lactobacillus plantarum microcapsule, lactobacillus rhamnosus microcapsule, lactobacillus plantarum microcapsule containing a biofilm and Lactobacillus rhamnosus microcapsule containing a biofilm prepared in example 1 were added, respectively, and were vibrated at a speed of 150rpm in a vibration incubator for gastric digestion, and sampled at 0, 60 and 120min, respectively. Gastric juice consisted of 0.02M PBS and 10mg/mL pepsin, and pH was adjusted to 2.0 with 1M HCl at 37 ℃.
During small intestine digestion, 2mL of the Lactobacillus plantarum suspension, lactobacillus rhamnosus suspension, lactobacillus plantarum microcapsule, lactobacillus rhamnosus microcapsule, lactobacillus plantarum microcapsule containing a biofilm and Lactobacillus rhamnosus microcapsule containing a biofilm prepared in example 1 were added to a test tube containing 8mL of simulated intestinal fluid, and the mixture was shaken at a speed of 150rpm in a shaking incubator for small intestine digestion, and sampled at 0, 120 and 240min, respectively. The small intestine digestive juice is 0.02M PBS,10mg/mL trypsin, 0.3wt% bile salt, 1M NaOH is used for adjusting pH to 6.8, and the digestion temperature is 37 ℃. The digested sample was centrifuged at 6000r/min for 5min to remove the digestate. The samples were resuspended in PBS buffer and the total number of colonies was determined by plate counting.
The shelf life survival rate testing method comprises the following steps: the lactobacillus plantarum/lactobacillus rhamnosus microcapsules containing the biological membrane are placed in a storage bottle for sealing treatment, stored at room temperature (25 ℃) and used for measuring the viable count of the probiotics by a plate colony count method after 45 days.
The survival rate of the two probiotics in the stomach digestion is shown in table 1, wherein the survival rate of the probiotics at 0min is taken as a reference.
TABLE 1
As can be seen from table 1, the free probiotics, the probiotic microcapsules and the probiotic microcapsules containing the biofilm show different decreasing trend of survival rate after digestion by simulated gastric fluid. At 120min, the survival rate of free lactobacillus plantarum was reduced to 33% and the survival rate of free lactobacillus rhamnosus was reduced to 46%. The decrease trend of the lactobacillus plantarum embedded by the microcapsule is greatly slowed down compared with that of the free lactobacillus plantarum. At 120min, the survival rate reaches 51%. In addition, lactobacillus rhamnosus also shows a phenomenon that the decrease in survival rate tends to be slowed down. Whereas microcapsules containing a biofilm showed the best results among the three samples. The biofilm-containing microencapsulated lactobacillus plantarum still showed 65% survival after simulated digestion for 120 min. Likewise, lactobacillus rhamnosus also shows the same trend. Thus, from the results, RG-I microcapsules can enhance the digestion resistance of probiotics in stomach, while microcapsules with biological membranes further improve the protection of the microcapsules against probiotics.
The survival rate of the two probiotics in the small intestine digestion is shown in table 2, wherein the survival rate of the probiotics at 0min is taken as a reference.
TABLE 2
As can be seen from table 2, the microcapsule-embedded probiotics significantly improved the survival rate of the probiotics in the small intestine, and both probiotics showed better effects. Meanwhile, the formation of the biological film can also improve the survival rate of the microcapsule in the small intestine. The invention has better improving effect on the survival rate of probiotics in the digestion process of the gastrointestinal tract.
After 45 days of storage at 25 ℃, the survival rates of free lactobacillus plantarum and lactobacillus rhamnosus were 45% and 59%, respectively, while the survival rates of the microcapsule-embedded lactobacillus plantarum and lactobacillus rhamnosus were 66% and 77%, respectively, which suggests that the microcapsules significantly improve their survival rates. The survival rate of the microcapsule with the biological film and the two bacteria is 74 percent and 83 percent respectively, which obviously improves the effect of the microcapsule, thus showing that the invention can obviously improve the storage stability of the probiotics.
Comparative example 1
Lactobacillus rhamnosus suspension, lactobacillus rhamnosus microcapsule and lactobacillus rhamnosus microcapsule containing a biofilm were prepared by the same method as in example 1, except that commercial pectin was used instead of RG-I pectin.
The encapsulation efficiency results show that the encapsulation efficiency of the lactobacillus rhamnosus microcapsule prepared in comparative example 1 is 45%.
The lactobacillus rhamnosus suspension, lactobacillus rhamnosus microcapsule and lactobacillus rhamnosus microcapsule containing a biological membrane prepared in comparative example 1 were tested by the same method as in example 2, and the survival rate in simulated gastric fluid is shown in table 3, wherein the survival rate of probiotics at 0min is taken as a reference.
TABLE 3 Table 3
As is clear from Table 3, the survival rate of free Lactobacillus rhamnosus was relatively large when the stomach digestion was simulated, and was 63% after 60min simulated digestion. And after 120min simulated digestion, the survival rate is less than half of 46%; after the microcapsule is embedded, the survival rate is slightly improved, and after simulated digestion for 120min, the survival rate is 53%; the microcapsule embedded probiotics with the biological film show better effect when being subjected to simulated digestion, and the survival rate of the probiotics can still reach 72 percent after 120 minutes of simulated digestion. The results show that the survival of the probiotics is slightly improved after embedding the probiotics according to the method of comparative example 1, but the microcapsule has poor tolerance to gastric digestion due to materials and manufacturing modes. The microcapsule is easy to be eroded by gastric acid, so that the structure of the microcapsule is damaged, the survival rate of probiotics is reduced, and the protective effect is obviously lower than that of the microcapsule prepared in the embodiment 1 of the invention.
Comparative example 2
Lactobacillus rhamnosus suspension, lactobacillus rhamnosus microcapsule and lactobacillus rhamnosus microcapsule containing a biological membrane are prepared by the same method as in example 1, except that sodium alginate is used for replacing RG-I pectin.
The encapsulation efficiency results show that the encapsulation efficiency of the lactobacillus rhamnosus microcapsule prepared in comparative example 2 is 63%.
The lactobacillus rhamnosus suspension, lactobacillus rhamnosus microcapsule and lactobacillus rhamnosus microcapsule containing a biological membrane prepared in comparative example 2 were tested for survival rate in simulated gastric fluid by the same method as in example 2, and the results are shown in table 4, wherein the survival rate of probiotics at 0min is taken as a reference.
TABLE 4 Table 4
From table 4 it can be seen that when free lactobacillus rhamnosus mimics gastric digestion, the survival rate is greatly affected because probiotics are less tolerant to acidity. After 120min of simulated gastric digestion, less than half of the survival rate was 46%; the microcapsule can provide a shell for probiotics, and has the effect of reducing the permeation of gastric acid, so that the survival rate of the probiotics is improved. Therefore, after the microcapsule is embedded, the survival rate is slightly improved, and after the stomach digestion is simulated for 120min, the survival rate is 54%; while the survival rate of the microcapsule-embedded probiotics with the biofilm is remarkably increased due to the generation of the biofilm when digestion is simulated. After 120min simulated digestion, the survival rate can still reach 72%. The results show that the survival of the probiotics is slightly improved after the probiotics are embedded according to the method of the comparative example 2, but the microcapsule taking sodium alginate as the raw material has poor tolerance to gastric acid, so that the survival rate is reduced, and the protection effect is obviously lower than that of the microcapsule prepared in the embodiment 1 of the invention.
The above embodiments are only illustrative of the preferred embodiments of the present invention and are not intended to limit the scope of the present invention, and various modifications and improvements made by those skilled in the art to the technical solutions of the present invention should fall within the protection scope defined by the claims of the present invention without departing from the design spirit of the present invention.

Claims (10)

1. The preparation method of the probiotic RG-I pectin microcapsule is characterized by comprising the following steps:
(1) Adding probiotic bacterial suspension and calcium carbonate solution into RG-I pectin water solution, and uniformly mixing to obtain mixed solution;
(2) Adding an oil phase containing an emulsifier into the mixed solution for emulsification reaction, then adding an oil phase containing glacial acetic acid, continuing to react, finally adding a sodium acetate solution, standing, separating to obtain a water phase, sieving, and washing to obtain the RG-I pectin microcapsule.
2. The method of claim 1, wherein in step (1), the concentration of RG-I pectin in the aqueous solution of RG-I pectin is 0.8 to 1.8wt%.
3. The method of claim 1, wherein in step (1), the mass ratio of RG-I pectin in the RG-I pectin aqueous solution to calcium carbonate in the calcium carbonate solution is 1:1-5.
4. The method of claim 1, wherein in step (2), the emulsifier comprises tween 80.
5. The method according to claim 4, wherein the volume fraction of the emulsifier in the oil phase containing the emulsifier is 0.5 to 2.5%.
6. The process according to claim 1, wherein in the step (2), after adding the glacial acetic acid-containing oil phase, the water-oil volume ratio in the reaction system is 1:1-5.
7. A probiotic RG-I pectin microcapsule prepared according to the method of any one of claims 1-6.
8. A method for preparing a probiotic RG-I pectin microcapsule containing a biological film, which is characterized by comprising the step of performing stimulated culture on the probiotic RG-I pectin microcapsule according to claim 7 to enable the probiotic bacteria embedded in the probiotic RG-I pectin microcapsule to form the biological film, so as to obtain the probiotic RG-I pectin microcapsule containing the biological film.
9. The method according to claim 8, wherein the calcium carbonate is added to the medium for stimulating the culture in an amount of 10 to 100mM.
10. A probiotic RG-I pectin microcapsule containing a biofilm prepared according to the preparation method of claim 8 or 9.
CN202310500051.XA 2023-05-06 2023-05-06 Probiotics RG-I pectin microcapsule and preparation method thereof Pending CN116508994A (en)

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