CN112869172A - Microcapsule wall material with probiotic specificity and preparation method and application thereof - Google Patents
Microcapsule wall material with probiotic specificity and preparation method and application thereof Download PDFInfo
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- 239000006041 probiotic Substances 0.000 title claims abstract description 75
- 235000018291 probiotics Nutrition 0.000 title claims abstract description 75
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- 239000000463 material Substances 0.000 title claims abstract description 57
- 230000000529 probiotic effect Effects 0.000 title claims abstract description 55
- 238000002360 preparation method Methods 0.000 title claims abstract description 18
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- 238000002156 mixing Methods 0.000 claims abstract description 13
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- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 description 1
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 description 1
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- 239000002609 medium Substances 0.000 description 1
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- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 description 1
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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
- A23L29/03—Organic compounds
- A23L29/045—Organic compounds containing nitrogen as heteroatom
-
- 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
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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
- A23L33/20—Reducing nutritive value; Dietetic products with reduced nutritive value
- A23L33/21—Addition of substantially indigestible substances, e.g. dietary fibres
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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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Abstract
The invention discloses a microcapsule wall material with probiotic specificity and a preparation method and application thereof, belonging to the technical field of probiotic preparations. The preparation method of the microcapsule wall material with probiotic specificity comprises the following steps: (1) mixing the isolated soy protein with oligosaccharide according to the mass ratio of 1: 1-3 to prepare a mixed solution with the protein concentration of 5-10% w/v; (2) uniformly stirring the mixed solution, hydrating overnight, and adjusting the pH value to 8.0; (3) heating the mixed solution with the adjusted pH value at 75-80 ℃ for 30-60 min to obtain a Maillard reaction product solution; (4) and (3) freeze-drying the Maillard reaction product solution to obtain the Maillard reaction product freeze-dried powder, namely the microcapsule wall material. The Maillard reaction product prepared from the soy protein isolate and the prebiotics oligosaccharide has the effects of inhibiting the growth of pathogenic bacteria and potential prebiotics, and has good probiotic pertinence when being used as a microcapsule wall material.
Description
Technical Field
The invention belongs to the technical field of probiotic preparations, and particularly relates to a microcapsule wall material with probiotic specificity, and a preparation method and application thereof.
Background
Probiotics are a class of active microorganisms that, when ingested in sufficient quantities, produce functional health benefits to the human body. The probiotics have the effects of adjusting intestinal flora, keeping intestinal health, preventing cardiovascular and cerebrovascular diseases, preventing oral diseases, regulating emotion, regulating organism immunity and the like. With the development of micro-ecological science and the improvement of the public on nutrition and health cognition, more and more consumers have consumed probiotic products as a health food or health care product. The data of the probiotic division of the Chinese food science and technology society shows that the consumption scale of probiotics in the Asia-Pacific region of 2016 accounts for 47% of the global proportion, wherein China is the largest probiotic consumption market in Asia, and has huge development prospects.
In recent years, probiotic products emerge endlessly, and the research is more intensive. Scientific research shows that probiotics is extremely sensitive to external adverse environment and is extremely easy to inactivate. During downstream processing, midstream transportation and storage and upstream gastrointestinal tract digestion of human bodies, probiotic products face a plurality of adverse factors such as temperature change, oxygen stress, gastric acid, digestive enzymes, bile salts, antibiotics and the like, and the adverse environmental conditions can influence the survival rate of the probiotics, so that the number of the probiotics reaching the intestinal tracts of the human bodies is less than the minimum concentration for exerting the probiotics effect. Therefore, it is of great significance to adopt certain technical means to improve the resistance of the probiotics to adverse environments.
At present, commonly used methods for improving stress resistance of probiotics include a protectant method, a preculture method, a resistant strain screening method and a microencapsulation method. Among them, the protective effect of the protective agent method and the pre-culture method is limited, the resistant strain screening method is complicated and tedious, the safety inspection is severe, the cost is high, and the microencapsulation method is favored due to the advantages of simple operation, low energy consumption, good protective effect and the like.
The micro-encapsulation technology of the probiotics refers to the technology of micro-encapsulating the probiotics by using a biocompatible wall material. In the food industry, microencapsulation of probiotics is an effective value-added technical means, and has the effects of stabilizing the activity of probiotics, prolonging the shelf life of products, and directionally transmitting and releasing.
The prebiotics are substances which can not be digested by the intestinal tract of a human body but can be selectively fermented by the probiotics, and have the beneficial effects of promoting the proliferation of the probiotics, improving the stress resistance of the probiotics, inhibiting the growth of pathogenic bacteria and the like. Prebiotics are often used as additives in probiotic microcapsules, and the use routes are generally two of the following: firstly, the microcapsule wall material is mixed with a microcapsule wall material for use, so that the compactness of a microcapsule wall material film is enhanced, and the protection effect of the microcapsule is improved; secondly, mixing the probiotic with the mixture to be used as synbiotics to prepare synbiotics microcapsules so as to enhance the stability of the probiotics. However, in either of the above-mentioned methods, the prebiotics are added to the probiotic microcapsules by physical means, and the protective effect on the probiotics is limited.
The prior probiotic microcapsule mostly adopts protein, polysaccharide or a mixture thereof as a wall material, the wall material has no probiotic specificity, and even if a certain amount of prebiotics are added in a physical mode in the microencapsulation process, the protection effect of the obtained probiotic microcapsule in the digestion process is still limited. Therefore, how to obtain a wall material with probiotic specificity, which can promote the proliferation of probiotics and selectively inhibit the growth of pathogenic bacteria, is a technical problem to be solved in the field.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provides a preparation method of a microcapsule wall material with probiotic specificity.
The invention also aims to provide the microcapsule wall material with probiotic specificity prepared by the preparation method.
The invention further aims to provide application of the microcapsule wall material with probiotic specificity.
The purpose of the invention is realized by the following technical scheme:
a preparation method of microcapsule wall material with probiotic specificity comprises the following steps:
(1) mixing the isolated soy protein with oligosaccharide according to the mass ratio of 1: 1-3 to prepare a mixed solution with the protein concentration of 5-10% (w/v);
(2) uniformly stirring the mixed solution, hydrating overnight, and adjusting the pH value to 8.0;
(3) heating the mixed solution with the adjusted pH value at the temperature of 75-80 ℃ for 30-60 min to obtain a Maillard reaction product solution;
(4) and (3) freeze-drying the Maillard reaction product solution to obtain Maillard reaction product freeze-dried powder, namely the microcapsule wall material with probiotic specificity.
The oligosaccharide of step (1) preferably comprises at least one of isomaltooligosaccharide, galactooligosaccharide and xylooligosaccharide.
The overnight hydration described in step (2) is preferably carried out in a refrigerator at 4 ℃.
The pH adjusting agent in the step (2) is preferably at least one of NaOH solution and HCl solution; more preferably at least one of a 1M NaOH solution and a 1M HCl solution.
A microcapsule wall material with probiotic specificity is prepared by the preparation method.
The microcapsule wall material with probiotic specificity is applied to preparation of probiotic microcapsules.
A probiotic microcapsule is prepared by preparing a wall material solution with a protein concentration of 8% -12% (w/v) from the microcapsule wall material with probiotic specificity, uniformly mixing the wall material solution with a bacterial suspension according to a ratio of 1: 5-10 (v/v), and adding gluconic acid-delta-lactone to obtain a mixed solution; emulsifying the mixed solution and soybean oil according to the proportion of 1: 3-5 (v: v), removing an oil phase, and washing to obtain the soybean oil.
The condition for uniform mixing is preferably 200-400 rpm stirring for 0.5-1 h; more preferably 300rpm for 1 h.
The addition amount of the gluconic acid-delta-lactone is preferably 2-4% (w/v) of the mixed solution.
The emulsifying condition is preferably 600-900 rpm stirring for 3-5 h; more preferably 900rpm for 3 hours.
The method for removing the oil phase is preferably removing the oil phase by filtration.
The washing is preferably performed with sterile physiological saline.
Compared with the prior art, the invention has the following advantages and effects:
(1) the soybean protein isolate has good dissolving, gelling, emulsifying, water holding and film forming capabilities, excellent processing characteristics, low price and easy obtainment, is a common plant protein wall material in the active substance microencapsulation process, can form an environment-friendly composite material together with other substances, and has huge development potential.
(2) Isomaltooligosaccharide, galacto-oligosaccharide, xylo-oligosaccharide and the like are prebiotics with demonstrated functionality and safety, and have a promoting effect on the growth of probiotics.
(3) The Maillard reaction is simple and easy to control, and the Maillard reaction product microcapsule wall material is easy to prepare.
(4) The Maillard reaction product prepared from the soy protein isolate and the prebiotics oligosaccharide has the effects of inhibiting the growth of pathogenic bacteria and potential prebiotics, and has good probiotic pertinence when being used as a microcapsule wall material.
Drawings
FIG. 1 is a graph of the results of the pathogen-inhibitory test of digested SPI-XOS in example 1; wherein, p <0.01, p <0.001, p < 0.0001.
FIG. 2 is a graph of the results of the experimental results of the potential prebiotic properties of the digested SPI-XOS in example 1.
FIG. 3 is a graph of the pathogen-inhibitory effect of digested SPI-IMO in example 2; wherein, p <0.01, p <0.001, p < 0.0001.
Figure 4 is a graph of the results of the experimental potential prebiotic properties of the digested SPI-IMO of example 2.
FIG. 5 is a graph showing the result of the pathogen-inhibitory test on digested SPI-GOS in example 3; wherein, p <0.01, p <0.001, p < 0.0001.
FIG. 6 is a graph of the results of the experimental potential prebiotic properties of the digested SPI-GOS of example 3.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but the present invention is not limited thereto.
The soybean protein isolate, the isomaltooligosaccharide, the galactooligosaccharide and the xylooligosaccharide are purchased from Shanghai-sourced leaf biotechnology limited; the soybean oil is first-grade goldfish soybean oil purchased from the Zhengfeng supermarket in the Tianhe area of Guangzhou city, Guangdong province; glucono-delta-lactone, pepsin, trypsin were purchased from bio-engineering (shanghai) gmbh; MRS broth, MRS agar, LB broth, LB agar, bile salts were purchased from Kyork, Inc., and the other reagents were analytically pure.
Lactobacillus casei was purchased from Yakult corporation (guangdong, guangzhou); the pathogenic bacteria are escherichia coli ATCC 25922 and staphylococcus aureus ATCC51650 which are purchased from Guangdong province microorganism strain preservation center;
the simulated gastric fluid is: NaCl 0.2% (w/v), pepsin (3000U/mg, pig source) 3.0g/L, pH 2.0; the simulated intestinal fluid is: dipotassium phosphate 0.68% (w/v), trypsin (250U/mg, pig source) 1.0g/L, bile salt 4.5g/L, pH 7.5.
Simulating gastrointestinal fluid digestion treatment: the simulated gastrointestinal fluid digestion treatment conditions are 37 ℃ and 50rpm, and after the simulated gastric fluid is treated for 2 hours, the simulated gastrointestinal fluid is transferred to the simulated intestinal fluid for treatment for 4 hours at 37 ℃ and 50 rpm.
The pathogenic bacteria inhibition detection method related in the embodiment is an Oxford cup method, wherein 100 mu L of microcapsule wall material digestive juice passes through a sterile water phase filter membrane with the thickness of 0.22 mu m, and then is added into the solution containing 10 percent (v/v) of 1 percent (v/v)6And (3) standing and absorbing the mixture for 12 hours at 4 ℃ in LB agar medium of CFU/mL escherichia coli ATCC 25922 or staphylococcus aureus ATCC51650, then transferring the mixture to an incubator at 37 ℃ for standing and culturing for 10 hours, recording the size of a bacteriostatic circle, and adopting 100 mu g/mL ampicillin as a positive control.
The potential prebiotics characteristic detection experimental method comprises the following steps: cheese milkThe bacillus is inoculated in an improved culture medium for culture, and OD is continuously monitored for 24h600nmA change in (c). The modified media used were: 2g/L of dipotassium phosphate, 2g/L of ammonium citrate, 3g/L of anhydrous sodium acetate, 0.2g/L of magnesium sulfate heptahydrate, 0.05g/L of manganese sulfate tetrahydrate and 45g/L of simulated digestion wall material sample, and adopting improved culture without digestion wall material as a blank control.
The probiotic detection method comprises the following steps: GB 4789.35-2016 lactic acid bacteria test for food safety national standard food microbiology inspection.
The method for measuring the embedding rate of the microcapsule comprises the following steps: determining the number of viable bacteria in the microcapsule and the total number of viable bacteria in the bacterial suspension added into the wall material;
the microcapsule embedding rate is calculated by the following formula:
the embedding rate (%) -, the number of live bacteria in the microcapsule/the total number of live bacteria in the bacterial suspension added into the wall material is multiplied by 100%.
Example 1
(1) A preparation method of microcapsule wall material with probiotic specificity comprises the following steps:
1) mixing Soybean Protein Isolate (SPI) and xylo-oligosaccharide (XOS) according to the mass ratio of 1:2 to prepare a mixed solution with the protein concentration of 5% (w/v);
2) uniformly stirring the mixed solution, hydrating the mixed solution in a refrigerator at 4 ℃ overnight, and then adjusting the pH of the mixed solution to 8.0 by using a 1M NaOH solution and a 1M HCl solution;
3) heating the mixed solution with the adjusted pH value at 80 ℃ for 30min to obtain a Maillard reaction product solution SPI-XOS;
4) freeze-drying the Maillard reaction product solution to obtain a Maillard reaction product SPI-XOS freeze-dried powder, namely the wall material with probiotic specificity;
(2)0.5g of SPI-XOS freeze-dried powder is sequentially added into 5mL of simulated gastric fluid and simulated intestinal fluid for simulated gastrointestinal fluid digestion treatment to obtain simulated digested SPI-XOS, and a pathogen (escherichia coli and staphylococcus aureus) inhibition experiment and a potential prebiotics characteristic detection experiment are carried out on the digested SPI-XOS.
As shown in FIGS. 1 and 2, it can be seen from FIG. 1 that SPI-XOS has a significant inhibitory effect on Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 51650; as can be seen from FIG. 2, SPI-XOS has a promoting effect on the growth of Lactobacillus casei.
(3) Inoculating Lactobacillus casei into MRS seed culture solution (MRS broth), and standing at 37 deg.C for 24 hr; inoculating MRS seed culture solution into 100mL of MRS culture solution (namely MRS broth) with the inoculation amount of 2% (v/v), and standing and culturing at 37 ℃ for 14 h; centrifuging the culture solution, collecting precipitate, washing, and dispersing Lactobacillus casei precipitate in 0.85% sterile physiological saline to obtain 10% concentration9CFU/mL of Lactobacillus casei suspension.
(4) Dissolving the Maillard reaction product SPI-XOS freeze-dried powder in sterile water to prepare a wall material solution with the protein concentration of 8% (w/v); mixing the lactobacillus casei suspension and the wall material solution in a ratio of 1:9(v: v), magnetically stirring the mixed solution at the rotating speed of 300rpm for 1h, and then adding 2% (w/v) gluconic acid-delta-lactone to obtain a mixed solution; then the mixed solution is emulsified with soybean oil at a ratio of 1:4(v: v), and magnetically stirred at 900rpm for 3h to form microcapsules; and finally removing the oil phase in a filtering mode, washing the obtained precipitate with sterile normal saline, washing away the residual oil phase, and finally obtaining the SPI-XOS/lactobacillus casei microcapsule which is the probiotic microcapsule, and detecting the embedding rate of the microcapsule.
(5)0.5g of probiotic microcapsules are sequentially added into 5mL of simulated gastric juice and intestinal juice to carry out simulated gastrointestinal fluid digestion treatment, and meanwhile, free lactobacillus casei is used as a control to detect the protection effect of the SPI-XOS wall material on the lactobacillus casei.
The results are shown in table 1 below:
table 1: protective effect of SPI-XOS wall material on lactobacillus casei
Example 2
(1) A preparation method of microcapsule wall material with probiotic specificity comprises the following steps:
1) mixing Soybean Protein Isolate (SPI) and isomaltose hypgather (IMO) according to the mass ratio of 1:2 to prepare a mixed solution with the protein concentration of 5% (w/v);
2) uniformly stirring the mixed solution, hydrating the mixed solution in a refrigerator at 4 ℃ overnight, and then adjusting the pH of the mixed solution to 8.0 by using a 1M NaOH solution and a 1M HCl solution;
3) heating the mixed solution with the adjusted pH value for 60min at the temperature of 80 ℃ to obtain a Maillard reaction product solution SPI-IMO;
4) freeze-drying the Maillard reaction product solution SPI-IMO to obtain the Maillard reaction product SPI-IMO freeze-dried powder, namely the wall material with probiotic specificity;
(2)0.5g of SPI-IMO freeze-dried powder is sequentially added into 5mL of simulated gastric fluid and simulated intestinal fluid for simulated gastrointestinal fluid digestion treatment to obtain simulated digested SPI-IMO, and a pathogen (escherichia coli and staphylococcus aureus) inhibition experiment and a potential prebiotics characteristic detection experiment are carried out on the digested SPI-IMO.
The results are shown in fig. 3 and 4, and it can be seen from fig. 3 that SPI-IMO has a significant inhibitory effect on escherichia coli ATCC 25922 and staphylococcus aureus ATCC 51650; as can be seen from FIG. 4, SPI-IMO has a promoting effect on the growth of Lactobacillus casei.
(3) Inoculating Lactobacillus casei into MRS seed culture solution (MRS broth), and standing at 37 deg.C for 24 hr; inoculating MRS seed culture solution into 100mL of MRS culture solution (namely MRS broth) with the inoculation amount of 2% (v/v), and standing and culturing at 37 ℃ for 14 h; centrifuging the culture solution, collecting precipitate, washing, and dispersing Lactobacillus casei precipitate in 0.85% sterile physiological saline to obtain 10% concentration9CFU/mL of Lactobacillus casei suspension.
(4) Dissolving the Maillard reaction product SPI-IMO freeze-dried powder in sterile water to prepare a wall material solution with the protein concentration of 8% (w/v); mixing the lactobacillus casei suspension and the wall material solution in a ratio of 1:9(v: v), magnetically stirring the mixed solution at the rotating speed of 300rpm for 1h, and then adding 2% (w/v) gluconic acid-delta-lactone to obtain a mixed solution; then the mixed solution is emulsified with soybean oil at a ratio of 1:4(v: v), and magnetically stirred at 900rpm for 3h to form microcapsules; and finally removing the oil phase in a filtering mode, washing the obtained precipitate with sterile normal saline, washing away the residual oil phase, and finally obtaining the SPI-IMO/lactobacillus casei microcapsule which is the probiotic microcapsule, and detecting the embedding rate of the microcapsule.
(5)0.5g of probiotic microcapsules are sequentially added into 5mL of simulated gastric juice and intestinal juice to carry out simulated gastrointestinal fluid digestion treatment, and meanwhile, free lactobacillus casei is used as a control to detect the protection effect of the SPI-IMO wall material on the lactobacillus casei.
The results are shown in table 2 below:
table 2: protective effect of SPI-IMO wall material on lactobacillus casei
Example 3
(1) A preparation method of microcapsule wall material with probiotic specificity comprises the following steps:
1) mixing Soybean Protein Isolate (SPI) and galacto-oligosaccharide (GOS) according to a mass ratio of 1:2 to prepare a mixed solution with a protein concentration of 5% (w/v);
2) uniformly stirring the mixed solution, hydrating the mixed solution in a refrigerator at 4 ℃ overnight, and then adjusting the pH of the mixed solution to 8.0 by using a 1M NaOH solution and a 1M HCl solution;
3) heating the mixed solution with the adjusted pH value for 60min at the temperature of 80 ℃ to obtain a Maillard reaction product solution SPI-GOS;
4) freeze-drying the Maillard reaction product solution to obtain a Maillard reaction product SPI-GOS freeze-dried powder, namely a microcapsule wall material with probiotic specificity;
(2)0.5g of SPI-GOS freeze-dried powder is sequentially added into 5mL of simulated gastric fluid and simulated intestinal fluid for simulated gastrointestinal fluid digestion treatment to obtain simulated digested SPI-GOS, and a pathogen (escherichia coli and staphylococcus aureus) inhibition experiment and a potential prebiotics characteristic detection experiment are carried out on the digested SPI-GOS.
As shown in FIGS. 5 and 6, it can be seen from FIG. 5 that the SPI-GOS has a significant inhibitory effect on Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 51650; as can be seen from FIG. 6, SPI-GOS has a promoting effect on the growth of Lactobacillus casei.
(3) Inoculating Lactobacillus casei into MRS seed culture solution (MRS broth), and standing at 37 deg.C for 24 hr; inoculating MRS seed culture solution into 100mL of MRS culture solution (namely MRS broth) with the inoculation amount of 2% (v/v), and standing and culturing at 37 ℃ for 14 h; centrifuging the culture solution, collecting precipitate, washing, and dispersing Lactobacillus casei precipitate in 0.85% sterile physiological saline to obtain 10% concentration9CFU/mL of Lactobacillus casei suspension.
(4) Dissolving a Maillard reaction product SPI-GOS freeze-dried powder in sterile water to prepare a wall material solution with the protein concentration of 8% (w/v); mixing the lactobacillus casei suspension and the wall material solution in a ratio of 1:9(v: v), magnetically stirring the mixed solution at the rotating speed of 300rpm for 1h, and then adding 2% (w/v) gluconic acid-delta-lactone to obtain a mixed solution; then the mixed solution is emulsified with soybean oil at a ratio of 1:4(v: v), and magnetically stirred at 900rpm for 3h to form microcapsules; and finally removing the oil phase in a filtering mode, washing the obtained precipitate with sterile normal saline, washing away the residual oil phase, and finally obtaining the SPI-XOS/lactobacillus casei microcapsule which is the probiotic microcapsule, and detecting the embedding rate of the microcapsule.
(5)0.5g of probiotic microcapsules are sequentially added into 5mL of simulated gastric juice and intestinal juice to carry out simulated gastrointestinal fluid digestion treatment, and meanwhile, free lactobacillus casei is used as a control to detect the protection effect of the SPI-GOS wall material on the lactobacillus casei.
The results are shown in table 3 below:
table 3: protective effect of SPI-GOS wall material on lactobacillus casei
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (10)
1. A preparation method of a wall material with probiotic specificity is characterized by comprising the following steps:
(1) mixing the isolated soy protein with oligosaccharide according to the mass ratio of 1: 1-3 to prepare a mixed solution with the protein concentration of 5-10% w/v;
(2) uniformly stirring the mixed solution, hydrating overnight, and adjusting the pH value to 8.0;
(3) heating the mixed solution with the adjusted pH value at the temperature of 75-80 ℃ for 30-60 min to obtain a Maillard reaction product solution;
(4) and (3) freeze-drying the Maillard reaction product solution to obtain Maillard reaction product freeze-dried powder, namely the microcapsule wall material with probiotic specificity.
2. The method according to claim 1, wherein the oligosaccharide of step (1) comprises at least one of isomaltooligosaccharide, galactooligosaccharide and xylooligosaccharide.
3. The method according to claim 1, wherein the pH adjusting agent in step (2) is at least one of NaOH solution and HCl solution.
4. A microcapsule wall material with probiotic specificity, which is prepared by the preparation method of any one of claims 1 to 3.
5. Use of a microcapsule wall material having probiotic specificity according to claim 4 for the preparation of probiotic microcapsules.
6. A probiotic microcapsule is characterized in that the microcapsule wall material with probiotic specificity in claim 4 is prepared into a wall material solution with protein concentration of 8% -12% w/v, then the wall material solution and bacterial suspension are uniformly mixed according to the proportion of 1: 5-10 v/v, and gluconic acid-delta-lactone is added to obtain a mixed solution; emulsifying the mixed solution and soybean oil at a ratio of 1: 3-5 v/v, removing an oil phase, and washing to obtain the soybean oil.
7. The probiotic microcapsule according to claim 6, wherein the mixing is performed under the condition of stirring at 200-400 rpm for 0.5-1 h.
8. The probiotic microcapsule according to claim 6, wherein the glucono-delta-lactone is added in an amount of 2% to 4% w/v of the mixed solution.
9. The probiotic microcapsule according to claim 6, wherein the emulsifying condition is 600-900 rpm stirring for 3-5 h.
10. The probiotic microcapsule according to claim 6, characterized in that the method for removing the oil phase is to remove the oil phase by filtration.
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