CN114209063A - Preparation method of fingered citron polysaccharide embedded lactobacillus paracasei microcapsule - Google Patents
Preparation method of fingered citron polysaccharide embedded lactobacillus paracasei microcapsule Download PDFInfo
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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
- A23L29/30—Foods or foodstuffs containing additives; Preparation or treatment thereof containing carbohydrate syrups; containing sugars; containing sugar alcohols, e.g. xylitol; containing starch hydrolysates, e.g. dextrin
-
- 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
- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
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- 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
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- Polymers & Plastics (AREA)
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Abstract
The invention discloses a preparation method of a fingered citron polysaccharide embedded lactobacillus paracasei microcapsule, which comprises the following steps: (1) preparing water phase solution from probiotic lactobacillus paracasei, fingered citron polysaccharide, sodium alginate, corn starch, calcium carbonate powder and water; (2) preparing an oil phase solution by using vegetable oil and an emulsifier; (3) mixing the water phase and the oil phase, and stirring to form a uniform water-in-oil solution; (4) adding acetic acid; (5) adding Tween 80 aqueous solution, stirring, standing, removing oil phase, filtering to obtain precipitate, washing to remove residual oil, and washing to obtain fructus Citri Sarcodactylis polysaccharide embedded Lactobacillus paracasei microcapsule. The invention adopts fingered citron polysaccharide, sodium alginate and starch to compound to form a compact embedding structure, effectively improves the embedding rate of lactobacillus paracasei, has good embedding effect, and the prepared microcapsule has the particle size distribution interval of 1.65-365.20 mu m, the volume average diameter D4, 3 is 136.5 mu m, and the median particle size D50 is 135.9 mu m.
Description
Technical Field
The invention relates to the technical field of food processing, in particular to a preparation method of a fingered citron polysaccharide embedded lactobacillus paracasei microcapsule.
Background
The probiotics is a general name of active microorganisms which are fixedly planted in the intestinal tract of a host and can generate definite beneficial effects by improving the microecological balance of the intestinal tract of the host, and has the effects of improving the digestibility of food, manufacturing nutrient substances, maintaining the balance of the flora in the intestinal tract, inhibiting the growth of pathogenic bacteria, further playing roles in enhancing nutrition, regulating immunity, preventing diseases and the like. Probiotics are often extremely sensitive to environmental conditions such as high temperature, oxygen, gastric acid, etc., which can severely limit their field of application and their beneficial health effects in humans. Therefore, effective embedding technical strategies are implemented on probiotics to improve the stability of the probiotics in food and human digestive tract, which is of great significance for improving the economic value of probiotic-rich products.
Microcapsules, which are micro-containers or packages having a polymer wall, have been widely used and studied as a technical means for effectively protecting probiotics against adverse environments. The principle of the microcapsule is that a compact physical barrier is formed around the probiotics, which is beneficial to resisting adverse conditions such as oxygen, gastric acid, bile salt and the like, thereby reducing the damage of adverse environment to the probiotics.
At present, common traditional embedding materials are mostly used for research in the technology of protecting probiotics by utilizing microcapsules, and novel health embedding materials are rarely developed. Microencapsulation is performed on probiotics in order to improve the resistance of the probiotics to adverse environments such as gastric acid, bile salt and the like, so that the probiotics still keep high activity when reaching the colon of a human body through the alimentary canal, the stomach, the small intestine and the like, and further play a probiotic role.
The Chinese invention applies for a preparation method of lactobacillus microcapsules, the application numbers are as follows: 201711288401.1, respectively; discloses a preparation method of a lactobacillus microcapsule: firstly, preparing synbiotic solution by using lactic acid bacteria and prebiotics; dissolving sodium alginate and pectin in distilled water, stirring at room temperature to obtain viscous mixed solution, and adding CaCO3Uniformly dispersing the powder in the mixed solution to obtain a microcapsule wall material, and uniformly mixing the microcapsule wall material with the synbiotic solution to obtain a bacterial glue mixed solution; and thirdly, preparing the microcapsule. The prepared lactic acid bacteria microcapsule can tolerate the action of gastric acid and bile salt so as to be better planted in intestinal tracts, plays the probiotic effects of various lactic acid bacteria such as immunoregulation, digestion improvement, tumor resistance, mutation resistance, acid-base balance regulation and the like, and simultaneously ensures that the prepared microcapsule has uniform particles with the particle size of about 100 mu m; the wall materials of sodium alginate and pectin used in the technology are common traditional materials, and the gastric acid and bile salt tolerance before and after embedding of probiotics is not compared. Therefore, at present, a need exists for developing a novel healthy wall material and applying the wall material to a probiotic microcapsule product, wherein the gastric acid and bile salt tolerance of the probiotic before and after being encapsulated is improved by comparing the gastric acid and bile salt tolerance of the probiotic before and after being encapsulated, so that the probiotic successfully colonizes the intestinal tract and plays a probiotic role.
Disclosure of Invention
The invention provides a preparation method of a fingered citron polysaccharide embedded lactobacillus paracasei microcapsule, aiming at the problems that the embedding wall material of probiotics microcapsules is mostly a common traditional material in the prior art, the resistance of the prepared microcapsule to adverse conditions such as gastric acid, cholate and the like is not enough, and the like, the preparation method of the fingered citron polysaccharide embedded lactobacillus paracasei microcapsule adopts traditional Chinese medicine fingered citron as a raw material, and through the steps of water extraction, alcohol precipitation and the like, a novel healthy wall material fingered citron polysaccharide is developed, has biological activities such as oxidation resistance, tumor resistance, immunoregulation and the like, and is successfully applied to embedding of the lactobacillus paracasei microcapsule, so that the microcapsule with strong gastric acid and cholate resistance and high stability is obtained.
In order to achieve the purpose, the technical scheme provided by the invention is as follows:
a preparation method of a fingered citron polysaccharide embedded lactobacillus paracasei microcapsule comprises the following operation steps:
(1) preparing probiotics lactobacillus paracasei, fingered citron polysaccharide, sodium alginate, corn starch, calcium carbonate powder and sterile water into an aqueous phase solution, wherein the mass concentration of probiotics in the aqueous phase solution is 10% -15%, the mass concentration of the fingered citron polysaccharide in the aqueous phase solution is 0.1% -1.5%, the mass concentration of sodium alginate in the aqueous phase solution is 0.6% -3%, the mass concentration of corn starch in the aqueous phase solution is 0.2% -1.8%, and the mass concentration of calcium carbonate powder in the aqueous phase solution is 0.1% -0.9%;
(2) preparing an oil phase solution by using vegetable oil and an emulsifier;
(3) mixing the water phase obtained in the step (1) and the oil phase obtained in the step (2), and stirring to form a uniform water-in-oil solution;
(4) adding glacial acetic acid into the water-in-oil solution obtained in the step (3) to initiate internal gel;
(5) adding 1% Tween 80 aqueous solution, slightly stirring to separate oil phase and water phase, standing, discarding oil phase, filtering to obtain precipitate (microcapsule), washing with 1% Tween 80 aqueous solution to remove residual oil, and washing with sterile water to obtain final product.
Preferably, the fingered citron polysaccharide in the step (1) is prepared by taking traditional Chinese medicinal materials of fingered citron as raw materials according to the mass ratio of fingered citron powder: adding water to the fingered citron powder at a ratio of 1:20, stirring and extracting at 80 ℃ for 5h, filtering and centrifuging to obtain a supernatant, carrying out rotary evaporation and concentration, precipitating with ethanol, adding ethanol to ensure that the final volume concentration is 80%, volatilizing the ethanol such as the precipitate, then re-dissolving with water, and freeze-drying to obtain fingered citron polysaccharide; the main monosaccharide composition of the obtained fingered citron polysaccharide is as follows: 63.62% of Man mannose, 24.16% of rha rhamnose, 10.62% of Gal-a galacturonic acid and 1.60% of Gal galactose.
Preferably, the volume concentration of the emulsifier in the oil phase solution in the step (2) is 2%.
Preferably, the vegetable oil of step (2) is soybean oil; the emulsifier is Tween 80.
Preferably, the stirring in step (3) is 400rpm for 15 min.
Preferably, the volume ratio of the addition amount of the glacial acetic acid to the water phase in the step (4) is 1: 250.
compared with the prior art, the invention has the following beneficial effects:
the microcapsule adopts the traditional Chinese medicine fingered citron as a wall material, and has the effects of biological activities such as oxidation resistance, tumor resistance, immunoregulation and the like; the invention adopts fingered citron polysaccharide, sodium alginate and starch to compound to form a compact embedding structure, effectively improves the embedding rate of lactobacillus paracasei, has good embedding effect, the embedding rate is as high as 94.33 percent, the particle size distribution interval of the prepared microcapsule is 1.65-365.20 mu m, the volume average diameter D [4,3] is 136.5 mu m, and the median particle size D50 is 135.9 mu m; furthermore, the number of viable bacteria of the microcapsule prepared by the invention is reduced by 1.33-1.45log after gastrointestinal fluid digestion is simulated, lactobacillus paracasei adopted by the invention has no activity after gastrointestinal fluid digestion is simulated under the same condition, and after embedding, the lactobacillus paracasei has good gastric acid resistance and cholate resistance, the application field of probiotics is widened, and the probiotics can better exert the probiotic effect.
Drawings
FIG. 1 is a particle size distribution diagram of the fingered citron polysaccharide-embedded lactobacillus paracasei microcapsule prepared by the invention.
FIG. 2 shows the particle size distribution interval and the accumulation rate of the fingered citron polysaccharide-embedded lactobacillus paracasei microcapsule prepared by the invention.
FIG. 3 is a photograph under a fluorescent microscope of Lactobacillus paracasei used in the present invention.
FIG. 4 is a picture under a fluorescent microscope of a fingered citron polysaccharide embedded lactobacillus paracasei microcapsule prepared by the invention; wherein (a) is a global view and (b) is a local focal map.
FIG. 5 is a picture of the solution for de-encapsulation under a fluorescence microscope after de-encapsulation of the fingered citron polysaccharide-embedded lactobacillus paracasei microcapsule prepared by the invention.
Detailed Description
The following detailed description is to be read in connection with the accompanying drawings, but it is to be understood that the scope of the invention is not limited to the specific embodiments. The raw materials and reagents used in the examples were all commercially available unless otherwise specified. In the examples, Lactobacillus paracasei is commercially available.
In the embodiment, the Tween 80 aqueous solution with the volume concentration of 1% is prepared by adding water to Tween 80; the Tween 80 and the Tween 80 for preparing the Tween 80 water solution with the volume concentration of 1% are the same substance; the volume concentration of the Tween 80 in the oil phase solution is 2 percent, which means the content of the Tween 80 in the oil phase.
The fingered citron polysaccharide adopted in the following embodiments adopts traditional Chinese medicinal materials fingered citron as raw materials, and the mass ratio of the fingered citron polysaccharide to the raw materials is as follows: adding water into the fingered citron powder at a ratio of 1:20, stirring and extracting for 5h at 80 ℃, filtering and centrifuging to obtain supernatant, carrying out rotary evaporation and concentration, precipitating by using ethanol, adding the ethanol to ensure that the final volume concentration is 80%, re-dissolving the obtained precipitate by using water after the ethanol is volatilized, and freeze-drying for 24h by using a vacuum freeze-drying machine to obtain fingered citron polysaccharide for later use; the main monosaccharide composition of the obtained fingered citron polysaccharide is as follows: 63.62% of Man mannose, 24.16% of rha rhamnose, 10.62% of Gal-a galacturonic acid and 1.60% of Gal galactose.
Example 1
A preparation method of a fingered citron polysaccharide embedded lactobacillus paracasei microcapsule comprises the following specific operation steps:
(1) preparing probiotic lactobacillus paracasei, the prepared fingered citron polysaccharide, sodium alginate, corn starch, calcium carbonate powder and sterile water into an aqueous phase solution, wherein the mass concentration of the probiotic in the aqueous phase solution is 10%, the mass concentration of the fingered citron polysaccharide in the aqueous phase solution is 0.1%, the mass concentration of the sodium alginate in the aqueous phase solution is 0.6%, the mass concentration of the corn starch in the aqueous phase solution is 0.2%, and the mass concentration of the calcium carbonate powder in the aqueous phase solution is 0.1%; the mass concentration is the content of the corresponding substance in the aqueous phase solution;
(2) preparing an oil phase solution by using soybean oil and Tween 80, wherein the volume concentration of the Tween 80 in the oil phase solution is 2%;
(3) mixing the water phase obtained in the step (1) and the oil phase obtained in the step (2), and stirring at 400rpm for 15min to form a uniform water-in-oil solution;
(4) adding glacial acetic acid into the water-in-oil solution obtained in the step (3) to initiate internal gel; the volume ratio of the addition amount of the glacial acetic acid to the water phase is 1: 250 of (a);
(5) adding 1% Tween 80 aqueous solution, slightly stirring to separate oil phase and water phase, standing, discarding oil phase, filtering to obtain precipitate (microcapsule), washing with 1% Tween 80 aqueous solution to remove residual oil, washing with sterile water to obtain Fingered citron polysaccharide embedded Lactobacillus paracasei microcapsule, and keeping the temperature at 4 deg.C in refrigerator.
Example 2
A preparation method of a fingered citron polysaccharide embedded lactobacillus paracasei microcapsule comprises the following specific operation steps:
(1) preparing probiotic lactobacillus paracasei, the prepared fingered citron polysaccharide, sodium alginate, corn starch, calcium carbonate powder and sterile water into an aqueous phase solution, wherein the mass concentration of the probiotic in the aqueous phase solution is 12.5 percent, the mass concentration of the fingered citron polysaccharide in the aqueous phase solution is 0.8 percent, the mass concentration of sodium alginate in the aqueous phase solution is 1.8 percent, the mass concentration of the corn starch in the aqueous phase solution is 1 percent, and the mass concentration of the calcium carbonate powder in the aqueous phase solution is 0.5 percent; the mass concentration is the content of the corresponding substance in the aqueous phase solution;
(2) preparing an oil phase solution by using soybean oil and Tween 80, wherein the volume concentration of the Tween 80 in the oil phase solution is 2%;
(3) mixing the water phase obtained in the step (1) and the oil phase obtained in the step (2), and stirring at 400rpm for 15min to form a uniform water-in-oil solution;
(4) adding glacial acetic acid into the water-in-oil solution obtained in the step (3) to initiate internal gel; the volume ratio of the addition amount of the glacial acetic acid to the water phase is 1: 250 of (a);
(5) adding 1% Tween 80 aqueous solution, slightly stirring to separate oil phase and water phase, standing, discarding oil phase, filtering to obtain precipitate (microcapsule), washing with 1% Tween 80 aqueous solution to remove residual oil, washing with sterile water to obtain Fingered citron polysaccharide embedded Lactobacillus paracasei microcapsule, and keeping the temperature at 4 deg.C in refrigerator.
Example 3
A preparation method of a fingered citron polysaccharide embedded lactobacillus paracasei microcapsule comprises the following specific operation steps:
(1) preparing probiotic lactobacillus paracasei, the prepared fingered citron polysaccharide, sodium alginate, corn starch, calcium carbonate powder and sterile water into an aqueous phase solution, wherein the mass concentration of the probiotic in the aqueous phase solution is 15%, the mass concentration of the fingered citron polysaccharide in the aqueous phase solution is 1.5%, the mass concentration of the sodium alginate in the aqueous phase solution is 3%, the mass concentration of the corn starch in the aqueous phase solution is 1.8%, and the mass concentration of the calcium carbonate powder in the aqueous phase solution is 0.9%; the mass concentration is the content of the corresponding substance in the aqueous phase solution;
(2) preparing an oil phase solution by using soybean oil and Tween 80, wherein the volume concentration of the Tween 80 in the oil phase solution is 2%;
(3) mixing the water phase obtained in the step (1) and the oil phase obtained in the step (2), and stirring at 400rpm for 15min to form a uniform water-in-oil solution;
(4) adding glacial acetic acid into the water-in-oil solution obtained in the step (3) to initiate internal gel; the volume ratio of the addition amount of the glacial acetic acid to the water phase is 1: 250 of (a);
(5) adding 1% Tween 80 aqueous solution, slightly stirring to separate oil phase and water phase, standing, discarding oil phase, filtering to obtain precipitate (microcapsule), washing with 1% Tween 80 aqueous solution to remove residual oil, washing with sterile water to obtain Fingered citron polysaccharide embedded Lactobacillus paracasei microcapsule, and keeping the temperature at 4 deg.C in refrigerator.
Performance detection：
And (3) detecting the embedding rate:
the specific determination method comprises the following steps: 500mg of the fingered citron polysaccharide-embedded lactobacillus paracasei microcapsule placed in a refrigerator for later use in each embodiment of the invention is suspended in 19.5mL of phosphate buffer (0.1M, pH 7.0) containing 2% alpha-amylase, and the suspension is uniformly mixed for 30 minutes at room temperature by using a vortex mixer, so that the lactobacillus paracasei in the fingered citron polysaccharide-embedded lactobacillus paracasei microcapsule is completely released; taking 1mL of the cyst-dissolving solution, diluting the cyst-dissolving solution by using sterile normal saline, inoculating the cyst-dissolving solution on an MRS solid culture medium, culturing the cyst-dissolving solution on a flat plate at 36 ℃ for 72 hours, and counting living cells; the embedding rate was calculated using the following formula:
embedding rate/% ([ viable cell count after embedding/viable cell count before embedding ] × 100
The result shows that the embedding rate of the fingered citron polysaccharide embedded lactobacillus paracasei microcapsule prepared by the invention can reach 94.33 percent (prepared in example 2) to the maximum extent, the embedding rate is high, and the embedding effect is good; the fingered citron polysaccharide, the sodium alginate and the starch are compounded to form a compact embedding structure, so that the embedding rate is effectively improved.
Simulating gastrointestinal fluid digestion:
artificial gastric juice treatment: weighing 500mg of the fingered citron polysaccharide embedded lactobacillus paracasei microcapsule prepared by the invention, placing the 500mg in 9.5mL of artificial gastric juice (pepsin 2000U/mL, pH 2.0), shaking for 2h at 37 ℃, immediately centrifuging at 10000rpm for 10min by using a centrifuge, and discarding the supernatant;
and (3) artificial intestinal juice treatment: adding 9.5mL of artificial intestinal juice (trypsin 100U/mL, bile salt with mass fraction of 0.3%, pH 8.0), shaking at 37 deg.C for 2 hr, centrifuging immediately (10000rpm, 10min), and removing supernatant;
and (3) capsule dissolution: adding 19.5mL of 0.1M phosphate buffer solution (containing 2% amylase) continuously for carrying out the cyst-lysis treatment;
and (3) culturing microorganisms: and (4) taking 1mL of the bacteria solution after the capsulization for gradient dilution, coating and counting.
The result shows that the viable count of the microcapsule prepared by the invention is reduced by 1.33-1.45log after the simulated gastrointestinal fluid digestion, but the lactobacillus paracasei adopted by the invention has no activity after the simulated gastrointestinal fluid digestion under the same condition, and the lactobacillus paracasei has good gastric acid resistance and cholate resistance after embedding. Meanwhile, compared with the microcapsule obtained by the technology of effective of resistant stage (Hi-mail) on the subvalval of bacteria infected with microorganisms with sodium alginate (after the microcapsule is treated for 360min by simulated gastrointestinal fluid, the viable count of ALGU (wet sodium alginate particle) and AHMU (wet sodium alginate + Hi-mail particle) is respectively reduced by 3.67log and 3.01 log), the microcapsule prepared by the invention has stronger gastric acid resistance and cholate resistance.
And (3) particle size detection:
the microcapsule particle size is detected by a Bettersize2600 laser particle size distribution instrument (wet method):
the results show that the microcapsule prepared by the invention has the following particle size distribution: the particle size distribution range is 1.65-365.20 μm, and the volume average diameter D [4,3] ═ 136.5 μm; d10 ═ 78.04 μm, D50 ═ 135.9 μm, D90 ═ 194.8 μm; SPAN (SPAN) 0.859;
note: d10: the cumulative particle distribution is 10% of the particle size, i.e. the volume content of particles smaller than this particle size is 10% of the total particles;
d50: the cumulative particle distribution is 50% of the particle size, also called median or median, which is a typical value representing the size of the particle, and which accurately divides the population into two equal parts, i.e. 50% of the particles exceed this value and 50% are below this value. If D50 of a sample is 5 μm, it indicates that of all the particle sizes constituting the sample, particles larger than 5 μm account for 50%, and particles smaller than 5 μm also account for 50%;
d90: the cumulative particle distribution is 90% of the particle size, i.e. the volume content of particles smaller than this particle size is 90% of the total particles;
SPAN (SPAN): SPAN (SPAN) is a descriptive value of the distribution width, for a symmetric distribution SPAN is 1; SPAN is more than 1, and large particles are large; SPAN is less than 1, and large particles are few.
The sodium Alginate microcapsule prepared by the prior art of Extending visual inspection of bismuth in Chitosan-Coated alumina Microcapsules Using Encapsulation and Internal deposition Encapsulation Technology has an average diameter of 154.3 μm and a span distribution of 1.15; the average particle size of the prepared sodium alginate chitosan microcapsule is 190.1 mu m, and the span distribution is 1.45; compared with the technology, the volume average diameter D4, 3 is 136.5 μm, SPAN (SPAN) is 0.859 < 1.000, and the large particles are few, which shows that the microcapsule prepared by the invention has uniform distribution and no overlarge particle size.
As can be seen from FIG. 3, the Lactobacillus paracasei used in the present invention is a Bacillus or a Bacillus longus. As can be seen from fig. 4(a), the circular outline of the microcapsules is visible under a fluorescence microscope; as can be seen from the partial focusing of the microcapsules in FIG. 4(b), the various wall materials are seen within the circular outline, in which they are embedded, indicating that the microcapsules produced form a dense structure in which Lactobacillus paracasei is embedded. As can be seen from FIG. 5, after the microcapsules were decapsulated, fragments of the microcapsules and released Lactobacillus paracasei rod were visualized under a fluorescence microscope.
The foregoing descriptions of specific exemplary embodiments of the present invention have been presented for purposes of illustration and description. It is not intended to limit the invention to the precise form disclosed, and obviously many modifications and variations are possible in light of the above teaching. The exemplary embodiments were chosen and described in order to explain certain principles of the invention and its practical application to enable one skilled in the art to make and use various exemplary embodiments of the invention and various alternatives and modifications as are suited to the particular use contemplated. It is intended that the scope of the invention be defined by the claims and their equivalents.
Claims (6)
1. A preparation method of a fingered citron polysaccharide embedded lactobacillus paracasei microcapsule is characterized by comprising the following operation steps:
(1) preparing probiotics lactobacillus paracasei, fingered citron polysaccharide, sodium alginate, corn starch, calcium carbonate powder and water into an aqueous phase solution, wherein the mass concentration of probiotics in the aqueous phase solution is 10% -15%, the mass concentration of the fingered citron polysaccharide in the aqueous phase solution is 0.1% -1.5%, the mass concentration of sodium alginate in the aqueous phase solution is 0.6% -3%, the mass concentration of corn starch in the aqueous phase solution is 0.2% -1.8%, and the mass concentration of calcium carbonate powder in the aqueous phase solution is 0.1% -0.9%;
(2) preparing an oil phase solution by using vegetable oil and an emulsifier;
(3) mixing the water phase obtained in the step (1) and the oil phase obtained in the step (2), and stirring to form a uniform water-in-oil solution;
(4) adding acetic acid into the water-in-oil solution obtained in the step (3);
(5) adding Tween 80 aqueous solution, stirring to separate oil phase and water phase, standing, removing oil phase, filtering to obtain precipitate, washing with Tween 80 aqueous solution to remove residual oil, and washing to obtain fructus Citri Sarcodactylis polysaccharide-embedded Lactobacillus paracasei microcapsule.
2. The method of claim 1, wherein: the fingered citron polysaccharide in the step (1) is prepared by taking traditional Chinese medicinal materials, namely fingered citron, as raw materials according to the mass ratio of fingered citron powder: adding water to the fingered citron powder at a ratio of 1:20, stirring and extracting at 80 ℃ for 5h, filtering and centrifuging to obtain a supernatant, carrying out rotary evaporation and concentration, precipitating with ethanol, adding ethanol to ensure that the final volume concentration is 80%, volatilizing the precipitated ethanol, redissolving with water, and freeze-drying to obtain fingered citron polysaccharide; the main monosaccharide composition of the obtained fingered citron polysaccharide is as follows: 63.62% of Man mannose, 24.16% of rha rhamnose, 10.62% of Gal-a galacturonic acid and 1.60% of Gal galactose.
3. The method of claim 1, wherein: the volume concentration of the emulsifier in the oil phase solution in the step (2) is 2%.
4. The method of claim 1, wherein: the vegetable oil in the step (2) is soybean oil; the emulsifier is Tween 80.
5. The method of claim 1, wherein: the stirring in the step (3) is 400rpm for 15 min.
6. The method of claim 1, wherein: the volume ratio of the addition amount of the acetic acid to the water phase in the step (4) is 1: 250.
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| CN105105104A (en) * | 2009-07-09 | 2015-12-02 | 德戈玛益生菌有限公司 | Heat resistant probiotic compositions and healthy food comprising them |
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| CN105105104A (en) * | 2009-07-09 | 2015-12-02 | 德戈玛益生菌有限公司 | Heat resistant probiotic compositions and healthy food comprising them |
| US20190029311A1 (en) * | 2015-12-17 | 2019-01-31 | Cj Cheiljedang Corporation | Coating method of lactic acid bacteria with increased intestinal survival rate |
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