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CN-121987572-A - Preparation and application of extracellular polysaccharide-based probiotics double-layer multi-effect microcapsule

CN121987572ACN 121987572 ACN121987572 ACN 121987572ACN-121987572-A

Abstract

The invention discloses a core-shell structure extracellular polysaccharide-based multi-effect probiotic microcapsule and a preparation method thereof. The microcapsule core is formed by crosslinking probiotics extracellular polysaccharide, vitamin B 3 and iron ions, and is embedded with probiotics, the shell is formed by compounding sodium alginate and the probiotics extracellular polysaccharide, and the microcapsule core is formed by dripping calcium chloride solution to carry out ionic gel curing. The method can obviously improve the encapsulation efficiency of the probiotics and endow the microcapsule with good pH response release characteristics, thereby effectively protecting the probiotics and realizing the targeted release of the probiotics in intestinal tracts. In addition, the extracellular polysaccharide of the probiotics in the system also has an anti-inflammatory effect, the vitamin B 3 can promote the metabolism of the probiotics to produce shikimic acid, the integral anti-inflammatory effect of the probiotics microcapsule is further enhanced, and the iron ions also have the physiological function of effectively preventing and improving anemia. The microcapsule obtained by the invention has wide application prospect in the fields of functional foods and probiotic preparations.

Inventors

  • LI ZHONGYUAN
  • WANG NAN
  • WANG XINRAN
  • WANG YANCHUN

Assignees

  • 天津科技大学

Dates

Publication Date
20260508
Application Date
20260211
Priority Date
20260210

Claims (8)

  1. 1. The preparation method of the extracellular polysaccharide-based probiotic microcapsule with the core-shell structure is characterized by comprising the following steps of: (1) Adding probiotics, vitamin B 3 solution, ferric chloride (FeCl 3 ) and Extracellular Polysaccharide (EPS) into deionized water, and uniformly stirring to obtain an inner core solution; (2) Adding sodium alginate (Alg) and Extracellular Polysaccharide (EPS) into deionized water, and uniformly stirring to obtain a shell solution; (3) Uniformly mixing the probiotic suspension with the inner core solution and the outer shell solution, adding the mixture into a calcium chloride (CaCl 2 ) solution by an extrusion method, standing to form gel microspheres, washing and drying to obtain the extracellular polysaccharide-based probiotic microcapsules with the core-shell structures.
  2. 2. The process according to claim 1, wherein the Extracellular Polysaccharide (EPS) is derived from Lactobacillus paracasei LH23 (Lacticaseibacillus paracasei) and has anti-inflammatory activity. The preservation number of the lactobacillus paracasei LH23 is CGMCC No. 16656, the preservation date is 10 months and 30 days in 2018, the biological institute of China academy of sciences No. 3 of West-1, ganyang region North Star of Beijing city, and the preservation unit is China general microbiological culture Collection center of China Committee.
  3. 3. The method for producing an extracellular polysaccharide according to claim 1, comprising the steps of: Inoculating activated strain into MRS liquid culture medium according to 2% inoculum size, culturing at 37deg.C for 24 h, centrifuging to obtain supernatant, concentrating supernatant, adding 3 times volume of absolute ethanol (final concentration 75%), standing at 4deg.C overnight, centrifuging at 8000 r/min for 10 min, and collecting precipitate; Preparing 5 mg/mL solution by precipitation, deproteinizing with trichloroacetic acid, standing at 4deg.C overnight, centrifuging at 8000 r/min for 20 min, collecting supernatant, dialyzing with dialysis bag with molecular weight cut-off of 8-14 kDa at 4deg.C for 48 h, changing water every 8 h, and lyophilizing the dialysate to obtain extracellular polysaccharide.
  4. 4. The preparation method according to claim 1, wherein the Extracellular Polysaccharide (EPS) solution in the step (1) has a concentration of 0.1-5% (w/v), the iron ion has a concentration of 5-20 mmol/L, and the stirring is carried out by a magnetic stirrer under stirring conditions of 400: 400 rpm and 20-30: 30 min at room temperature.
  5. 5. The method according to claim 1, wherein the concentration of sodium alginate (Alg) solution in step (2) is 0.5% -2% (w/v), and the concentration of Extracellular Polysaccharide (EPS) solution is 0.1% -5% (w/v).
  6. 6. The preparation method of claim 1, wherein in the step (3), the probiotic suspension, the core solution and the shell solution are uniformly mixed, added into 0.1% -5% (w/v) CaCl 2 solution by an extrusion method, and kept stand to form gel microspheres, and the gel microspheres are washed and dried to obtain the microcapsules.
  7. 7. The method according to claim 1, wherein the concentration of the probiotic bacteria liquid in step (1) is 10 7 -10 10 CFU/mL, the centrifugation speed is usually 8000-10000 rpm, and the time is usually 10-15 min.
  8. 8. The method for preparing the embedded microcapsule of claim 1, the embedded probiotics are one or more of Bifidobacterium adolescentis, bifidobacterium animalis ((Lactobacillus bifidus), bifidobacterium bifidum, bifidobacterium breve, bifidobacterium infantis, bifidobacterium longum, lactobacillus acidophilus, lactobacillus casei, lactobacillus crispatus, lactobacillus delbrueckii subspecies bulgaricus (Lactobacillus bulgaricus), lactobacillus delbrueckii subspecies lactis, lactobacillus fermentum, lactobacillus gasseri, lactobacillus helveticus, lactobacillus johnsonii, lactobacillus paracasei, lactobacillus plantarum, lactobacillus reuteri, lactobacillus rhamnosus, lactobacillus salivarius and Streptococcus thermophilus.

Description

Preparation and application of extracellular polysaccharide-based probiotics double-layer multi-effect microcapsule Technical Field The invention belongs to the technical field of probiotics delivery and core-shell type microcapsules, and relates to a core-shell structure extracellular polysaccharide-based probiotics microcapsule and a preparation method thereof. Background Probiotics are active microorganisms which have a positive effect on host health, but are highly sensitive to external environments (such as high temperature, high salt, gastric acid and bile acid), are easy to inactivate during processing, storage and gastrointestinal tract transportation, and are difficult to achieve the minimum viable bacteria amount required by intestinal tract colonization, so that the application range and the probiotic efficacy are severely limited. The microcapsule technology constructs a physical barrier through natural or synthetic polymer materials, so that probiotics can be effectively protected from adverse environmental stress, the survival rate of the probiotics in the processing, storage and digestion processes is improved, and the targeted release and intestinal field planting of the probiotics are realized. The wall materials of the probiotics microcapsule are mainly divided into polysaccharides, proteins and lipids, wherein the polysaccharide wall materials are mainly selected because of abundant sources, safety, no toxicity and good biocompatibility. Sodium alginate is mostly adopted as a wall material in the existing microcapsule preparation, and although the sodium alginate has the advantages of good biocompatibility, quick gel formation, easily available raw materials and the like, the sodium alginate has the defects of poor structural stability, environmental sensitivity, difficult controlled release, weak process controllability, limited industrialization and the like, and has room for improvement in key performances such as survival rate, long-term storage stability and the like in simulated gastrointestinal tract environments. The probiotics Extracellular Polysaccharide (EPS) is water-soluble polysaccharide which is metabolized and secreted by probiotics, can be crosslinked with cations in molecules to form microgels, has gelling property, emulsifying property and film forming property, can be used as prebiotics to promote the growth of the probiotics, and can form synergistic probiotics with the probiotics. Therefore, the EPS is added into the sodium alginate microcapsule system, so that the mechanical strength and swelling resistance of the microcapsule are improved, the acid resistance is improved, the gastric acid erosion is isolated, the microcapsule is prevented from being disintegrated in advance in the stomach, the active ingredients are ensured to smoothly reach the intestinal tract, the leakage of water-soluble ingredients is reduced, the embedding rate of the composite system is improved, and the like, so that the availability of the microcapsule is improved. Shikimic acid produced by probiotics has anti-inflammatory effect, and mainly plays an effect through multi-target mechanisms such as regulating and controlling inflammatory signal channels, inhibiting inflammatory mediator release, resisting oxidative stress and the like. Vitamin B 3（VB3) can further enhance the shikimate synthesis ability of the cells. In addition, iron plays an indispensable role in various core physiological functions in organisms, and its main mechanism of action is to integrate as a key prosthetic group into various functional proteins through reversible redox reactions. These functions cover the binding, transport and storage of oxygen, in particular as core component of hemoglobin and myoglobin, directly involved in the blood-replenishing process and improving iron deficiency anemia, and in addition it is involved in electron transfer and bioenergy synthesis in the cellular respiratory chain, as well as in a number of basic biological processes such as genetic material synthesis, immune response regulation and neural signaling as an essential cofactor. The maintenance of iron homeostasis is critical for the normal metabolism, growth and development, physiological function of the living body, and effective prevention and improvement of the anemic state. Therefore, a microcapsule system with a 'kernel-shell' dual structure is constructed, wherein the kernel is formed by the cross-linking of the extracellular polysaccharide of the probiotics, the vitamin B 3 and the iron ions, and the shell is formed by the combination of the extracellular polysaccharide of the probiotics and the sodium alginate. The double-layer design microcapsule not only can effectively improve the storage stability and the intestinal targeted delivery efficiency of the encapsulated probiotics, but also endows the microcapsule with multiple physiological regulation functions of anti-inflammatory, anemia improvement and the like, and has