CN117482157A - Probiotic preparation with high activity and high stability and preparation method thereof - Google Patents
Probiotic preparation with high activity and high stability and preparation method thereof Download PDFInfo
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- CN117482157A CN117482157A CN202311417368.3A CN202311417368A CN117482157A CN 117482157 A CN117482157 A CN 117482157A CN 202311417368 A CN202311417368 A CN 202311417368A CN 117482157 A CN117482157 A CN 117482157A
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- 239000006041 probiotic Substances 0.000 title claims abstract description 52
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- 238000000034 method Methods 0.000 claims abstract description 23
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- 239000012153 distilled water Substances 0.000 claims abstract description 12
- 238000002791 soaking Methods 0.000 claims abstract description 9
- 238000002156 mixing Methods 0.000 claims abstract description 6
- 235000013305 food Nutrition 0.000 claims abstract description 5
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 31
- 239000000843 powder Substances 0.000 claims description 23
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- 239000000047 product Substances 0.000 claims description 17
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
-
- 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
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/77—Sapindaceae (Soapberry family), e.g. lychee or soapberry
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/10—Laxatives
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/14—Prodigestives, e.g. acids, enzymes, appetite stimulants, antidyspeptics, tonics, antiflatulents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
-
- 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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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
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Abstract
The invention discloses a method for preparing a probiotic preparation by utilizing pineapple peel and soapberry fermentation, belonging to the fields of food science and biotechnology. The method comprises the following operation steps: firstly preparing pineapple peel fermentation liquor and soapberry fermentation liquor, respectively soaking pineapple peel and soapberry in distilled water, adding saccharomycetes for fermentation and filtering to obtain pineapple peel fermentation liquor and soapberry fermentation liquor; mixing the two fermentation liquids, adding brown sugar, filtering, adding saccharomycetes, and standing for 36-63 h to complete secondary fermentation, thus preparing the probiotic preparation. The method can improve survival rate and colonization ability of probiotics. The method is simple and easy to implement, can fully utilize the waste resources such as pineapple peel, soapberry and the like, and meanwhile, the prepared probiotic preparation has higher activity and stability.
Description
Technical Field
The invention relates to a probiotic preparation with high activity and stability and a preparation method thereof, belonging to the fields of food science and biotechnology.
Background
The intestinal flora imbalance is caused by the factors such as dietary structure change, environmental pollution, pressure increase and the like in modern life, and the immunity, digestive system function and nutrition absorption of a human body are negatively influenced. In order to regulate the balance of intestinal flora, enhance immunity and improve intestinal health, probiotic preparations are needed. The intestinal tract is the largest microbial ecosystem in humans, which contains a large number of bacterial populations, including beneficial and harmful bacteria. Probiotics are a kind of microorganisms beneficial to human bodies, and can help to maintain intestinal flora balance, promote digestion and absorption, enhance immunity and prevent and improve various digestive system related diseases.
The journal paper "classification of probiotics and its multi-field application research status quo" mentions that along with the continuous deep and development of research, the application field of probiotics is continuously expanding, but in practical application, the problems of poor stability of strains, difficult colonisation in digestive tract, unclear action mechanism for exerting certain efficacy and the like still exist. For example: the live bacteria preparation is easy to inactivate in the processes of feed processing, transportation and storage, the survival rate of the live bacteria preparation is not ensured, the biological activity is reduced, and the quality of the product is affected; most probiotics entering the digestive tract of animals are influenced by gastric acid, bile salts and the like, so that the quantity of viable bacteria which can finally reach the intestinal tract and colonize to play a role is insufficient, and the probiotic effect is not obvious. In addition, probiotics have certain toxicity problems while playing a role, and most lactobacillus and bifidobacteria produce various bacteriocins in the metabolic process, and the bacteriocins can inhibit or kill saprophytic bacteria and pathogenic bacteria in intestinal tracts and act on other microorganisms in the intestinal tracts, which are beneficial to maintaining the normal physiological functions of organisms. Finally, the preparation process of the probiotic preparation is complex, the production cost is high, and the product price is unstable and the popularity rate is limited. Therefore, a new solution is needed to improve the above-mentioned problems of conventional probiotic formulations to improve the quality, efficacy, safety and economy of the product.
The invention discloses a pineapple dreg packaging and fermenting feed and an energy-saving processing method, wherein pineapple dreg is taken as a main raw material, and the pineapple dreg is processed into special fermented feed for ruminants such as dairy cows, deer, pigs and the like through the procedures of crushing and juicing, adding dreg and a fermenting agent, quick primary drying, profiling bagging, sealing packaging, solid-state fermentation and the like. The invention has simple process, less equipment investment, low cost, changing waste into valuables, small volume, large density of finished fermented feed products, regular and beautiful shape, fresh-keeping storage and long-distance transportation, and realizing fresh-keeping storage and transportation and commercialization of pineapple residues. Patent CN 102150746a discloses a deep processing method of pineapple peel and slag. After pineapple is used for producing fruit juice or preserved fruit, the produced fruit peel and fruit residues are used as waste, and are usually used as auxiliary feed for livestock cultivation, and the coarse low-level treatment mode does not have any treatment, so that huge waste of resources is caused, and simultaneously, larger pressure is also caused to the environment. The invention prepares the fermented feed and the probiotic preparation by utilizing the specific technical steps of rhizopus, candida, lactobacillus, active enzyme and EM raw juice, performing primary fermentation, secondary fermentation, squeezing, fermentation juice treatment and the like on pineapple peel and residues. The detection shows that the crude protein content of the fermented feed reaches 23%, and the crude protein content after washing reaches 16%. The probiotics preparation can purify water quality, has the effects of promoting growth of animals, enhancing immunity and disease resistance of the animals, and the like, is a product for replacing antibiotics in the breeding industry, and has good use effect in cattle, sheep and aquaculture. Both patents disclose methods for preparing a probiotic preparation by fermenting pineapple peels and residues, but the probiotic preparation prepared by fermenting pineapple peels or residues has low probiotic activity, poor stability and corresponding hidden danger in long-term safety.
Disclosure of Invention
The invention aims to solve the problems of easy inactivation, poor stability, certain toxicity, complex preparation process, high production cost and the like of the traditional probiotic preparation, and provides a preparation method of the probiotic preparation.
In view of the above-mentioned objects, the present invention provides a method for preparing a probiotic preparation having high activity and high stability, comprising the steps of:
(1) Mixing pineapple peel fermentation liquor and soapberry fermentation liquor together, adding brown sugar, and stirring to obtain a mixed solution;
(2) Filtering the mixed solution prepared in the step (1) to obtain filtrate;
(3) Pouring the filtrate obtained in the step (2) into a fermentation tank, adding saccharomycetes, stirring until the saccharomycetes are completely dissolved, and standing and fermenting for 36-63 h to obtain the probiotic preparation.
In one embodiment of the present invention, in the step (1), the volume ratio of pineapple peel fermentation liquor to soapberry fermentation liquor is 1:3-3:1.
In one embodiment of the present invention, in the step (1), the mass ratio of the brown sugar to the mixed liquid of pineapple peel fermentation liquid and soapberry fermentation liquid is 1:3-1:5.
In one embodiment of the present invention, in the step (1), the preparation method of the pineapple peel fermentation broth includes the following steps:
a1. cleaning fresh pineapple peel, peeling, and cutting into small pieces;
b1. putting the pineapple peel small pieces in the step a1 into a fermentation tank, adding distilled water, and soaking to obtain a solution to be fermented;
c1. adding saccharomycetes into the solution to be fermented, standing and fermenting for 36-63 h, and filtering to obtain pineapple peel fermentation liquor.
In one embodiment of the present invention, the size of the small block in step a1 is in the range of 7mm×7mm to 11mm×11mm.
In one embodiment of the present invention, in step b1, the feed liquid ratio of pineapple peel pieces to distilled water is 1: 4-1: 9.
in one embodiment of the present invention, in step b1, the soaking time is 35min to 55min.
In one embodiment of the invention, in step c1, the yeast used is a cultured yeast strain, and the culturing method is that yeast powder is added to an HS culture medium for culturing, wherein the components of the HS culture medium comprise 4g/L-6g/L of bacteriopeptone, 19g/L-21g/L of glucose, 4g/L-6g/L of yeast extract, 2.6g/L-2.8g/L of dibasic sodium phosphate, 1g/L-1.3g/L of citric acid and 0.5g/L-1.5g/L of magnesium sulfate.
In one embodiment of the present invention, in step c1, the culturing conditions of the yeast are: the pH value is 2.3-5.2, the temperature is 20-29 ℃, and the culture time is 36-80 h, preferably 36-63 h.
In one embodiment of the present invention, in step c1, the yeast powder is added in an amount of 0.8% -2% by mass of the HS medium when culturing the yeast.
In one embodiment of the invention, in step c1, the yeast is added in an amount of 26% -37% of the volume of the solution to be fermented.
In one embodiment of the invention, in step c1, the temperature at which the fermentation is carried out is from 20℃to 29℃and the pH is from 2.3 to 5.2.
In one embodiment of the present invention, in step c1, the mesh size of the screen at the time of filtration is 50 mesh.
In one embodiment of the present invention, in the step (1), the preparation method of the soapberry fermentation broth includes the following steps:
a2. washing soapberry and grinding the soapberry into fine powder, putting the obtained soapberry powder into a fermentation tank, adding distilled water, and soaking the soapberry powder to obtain a solution to be fermented;
b2. adding saccharomycetes into the solution to be fermented in the step a2, standing for fermentation, and filtering to obtain soapberry fermentation liquor.
In one embodiment of the present invention, the fine powder of soapberry powder in step a2 has a particle size ranging from 95 mesh to 137 mesh.
In one embodiment of the present invention, in step a2, the ratio of soapberry powder to distilled water is 1: 4-1: 9.
in one embodiment of the present invention, the soaking time in step a2 is 35min to 55min.
In one embodiment of the invention, in step b2, the yeast used is a cultured yeast strain, and the culturing method is that yeast powder is added to an HS culture medium for culturing, wherein the components of the HS culture medium comprise 4g/L-6g/L of bacteriopeptone, 19g/L-21g/L of glucose, 4g/L-6g/L of yeast extract, 2.6g/L-2.8g/L of dibasic sodium phosphate, 1g/L-1.3g/L of citric acid and 0.5g/L-1.5g/L of magnesium sulfate.
In one embodiment of the present invention, in step b2, the culturing conditions of the yeast are: the pH value is 2.3-5.2, the temperature is 20-29 ℃, and the culture time is 36-80 h, preferably 36-63 h.
In one embodiment of the present invention, in step b2, the yeast powder is added in an amount of 0.8% -2% by mass of the HS medium when culturing the yeast.
In one embodiment of the invention, in step b2, the yeast is added in an amount of 26% -37% of the volume of the solution to be fermented.
In one embodiment of the invention, in step b2, the temperature at which the fermentation is carried out is 20℃to 29℃and the pH is 2.3 to 5.2.
In one embodiment of the invention, in step b2, the screen mesh size upon filtration is 50 mesh.
In one embodiment of the present invention, in step (2), the mesh size of the screen at the time of filtration is 70 mesh.
In one embodiment of the invention, in the step (3), the yeast is a cultured yeast strain, and the culturing method is that yeast powder is added to an HS culture medium for culturing, wherein the components of the HS culture medium comprise 4g/L to 6g/L of bacteriopeptone, 19g/L to 21g/L of glucose, 4g/L to 6g/L of yeast extract, 2.6g/L to 2.8g/L of dibasic sodium phosphate, 1g/L to 1.3g/L of citric acid and 0.5g/L to 1.5g/L of magnesium sulfate.
In one embodiment of the present invention, in step (3), the culturing conditions of the yeast are: the pH value is 2.3-5.2, the temperature is 20-29 ℃, and the culture time is 36-80 h, preferably 36-63 h.
In one embodiment of the present invention, in step (3), the yeast powder is added in an amount of 0.8% -2% by mass of the HS medium when the yeast is cultured.
In one embodiment of the invention, in step (3), the yeast is added in an amount of 26% -37% of the filtrate volume.
In one embodiment of the invention, in step (3), the temperature at which fermentation is carried out is 20℃to 29℃and the pH is 2.3 to 5.2.
The invention also provides a probiotic preparation prepared by the method.
The invention also provides application of the probiotic preparation in preparing foods, medicines and health care products.
In one embodiment of the invention, the health care product comprises a health care product with the functions of enhancing immunity, regulating intestinal flora, promoting digestion and relaxing bowels.
The invention has the beneficial effects that:
(1) Improving the activity of the probiotic preparation: enzymes, strains and active ingredients in pineapple peel and soapberry can promote the growth and metabolism of probiotics, so that the activity of the preparation is improved.
(2) Improving the stability of the probiotic preparation: solid residues and impurities are removed through steps of mixed fermentation, extraction and the like, so that a pure probiotic preparation is obtained, and the stability and the shelf life of the probiotic preparation are improved.
(3) Toxicity is reduced: the invention adopts the natural pineapple peel and soapberry as raw materials, and has lower toxicity and side effect compared with chemical synthesized components, and is safer and more reliable.
(4) The preparation process is simplified: the preparation method is relatively simple, does not need complex equipment and operation, and reduces the complexity and production cost of the preparation process.
Detailed Description
The following description of the preferred embodiments of the present invention is provided for better illustration of the invention, and should not be construed as limiting the invention.
The present invention to evaluate the effect of the different examples and comparative examples, the following tests were carried out:
1. validity test
The effectiveness test is carried out by referring to the detection methods of GB 4789.34 and GB 4789.35 in the national health care society standard "Probiotics viable count general rule", and comprises the test of the number of the probiotics and the test of the activity of the probiotics.
2. Stability test
The stability test is carried out according to the specification of the original food and drug administration, three-function [2013]500, notification of the food and drug administration office on the guidelines of stability test of printed health food. Including temperature stability testing, humidity stability testing, and acid and alkali resistance stability testing.
3. Safety test
Safety tests were performed with reference to the work of the safety evaluation procedure for food-grade strains for national food safety standards, established by the national Committee for Wei Jian, including acute toxicity tests, long-term toxicity tests, immunogenicity tests and drug resistance tests.
Example 1:
a method for preparing a probiotic preparation with high activity and high stability, comprising the steps of:
1. preparation of pineapple peel fermentation liquor
(1) The fresh pineapple peel is cleaned, peeled and cut into small pieces, and the specifications of the small pieces are 7mm multiplied by 7mm.
(2) Putting pineapple peel small pieces into a fermentation tank, adding distilled water, and soaking for 50min, wherein the feed liquid ratio of the pineapple peel small pieces to the distilled water is 1: and 7, obtaining a solution to be fermented.
(3) Adding saccharomycetes into the fermentation tank, wherein the adding amount of the saccharomycetes accounts for 37% of the volume of the solution to be fermented in the step (2); standing, fermenting pineapple peel to obtain a fermentation product, wherein the temperature during fermentation is 26 ℃, the pH value is 4.8, and the fermentation time is 63h;
(4) Filtering the fermentation product obtained in the step (3), filtering pineapple peel, and leaving fermentation liquor, wherein the mesh size of a filter screen is 50 meshes during filtering.
2. Preparation of soapberry fermentation broth
(1) Washing soapberry and grinding into fine powder for standby, wherein the particle size of the fine powder is 137 meshes;
(2) Placing soapberry powder into a fermentation tank, adding distilled water, and soaking for 50min; the feed liquid ratio of the soapberry fine powder to the distilled water is 1: and 7, obtaining a solution to be fermented.
(3) Adding saccharomycetes into the fermentation tank, wherein the strain content accounts for 37% of the volume of the solution to be fermented in the step (2); standing, and fermenting soapberry to obtain a fermentation product, wherein the temperature during fermentation is 26 ℃, the pH value is 4.8, and the fermentation time is 63h;
(4) Filtering the fermentation product obtained in the step (3), and filtering soapberry to leave fermentation liquor, wherein the mesh size of a filter screen is 50 meshes during filtering.
3. Preparation of probiotic formulations
(1) The pineapple peel fermentation liquor and soapberry fermentation liquor are mixed according to the volume ratio of 3:2 mixing together and adding brown sugar; the mass ratio between the brown sugar and the mixed solution is 1:4.
(2) The brown sugar was completely dissolved in the mixed solution by using a stirrer, and then poured into a filter, and the mixture was filtered using a sieve having a mesh size of 70 mesh.
(3) Pouring the filtered juice into a fermentation tank, and adding saccharomycetes; the strain content is 37% of the volume of the filtered juice, and the juice is left to ferment after being stirred uniformly, so as to obtain the probiotic preparation. The temperature during fermentation is 26 ℃, the pH value is 4.8, and the fermentation time is 63h.
In example 1, the yeasts are cultured yeasts, and the culturing process of the yeasts is as follows: adding yeast powder to an HS culture medium for culture, wherein the components of the HS culture medium comprise 5.2g/L of bacteriopeptone, 20.2g/L of glucose, 5.2g/L of yeast extract, 2.75g/L of dibasic sodium phosphate, 1.2g/L of citric acid and 1.1g/L of magnesium sulfate; the addition amount of the yeast powder during culture is 1.2% of the mass of the HS culture medium; the culture conditions of the saccharomycetes are as follows: the pH value is 4.8, the temperature is 26 ℃, and the culture time is 63h. Wherein the yeast powder is from Angel Yeast Co., ltd, and has a specification of 5 g/bag; HS culture medium is from Jiangsu next philosophy instruction instrument Co., ltd, and has the specification of 100mm.
Example 2:
example 2 differs from example 1 in that the cultivation time of the yeasts is 80h.
Comparative example 1:
comparative example 1 differs from example 1 in that the mixing ratio of pineapple peel fermentation broth and soapberry fermentation broth was changed to 7:1.
comparative example 2:
comparative example 2 differs from example 1 in that the pH value in the fermentation conditions at the time of preparation of the probiotic preparation was adjusted to 7.0.
Comparative example 3:
comparative example 3 differs from example 1 in that in step 3, brown sugar was not added.
Comparative example 4:
comparative example 4 differs from example 1 in that the temperature in the fermentation conditions at the time of preparation of the probiotic preparation was adjusted to 50 deg.c
Comparative example 5:
comparative example 5 differs from example 1 in that step 2 was omitted and step 3 did not contain soapberry fermentation broth.
Comparative example 6:
comparative example 6 differs from example 1 in that step 1 was omitted and step 3 did not contain pineapple peel broth
Comparative example 7:
comparative example 7 differs from example 1 in that step 3 was omitted, and the pineapple peel fermentation broth obtained in step 1 and the soapberry fermentation broth obtained in step 2 were directly mixed, without performing a secondary fermentation operation.
TABLE 1 results of effectiveness test of probiotic formulations prepared in examples 1-2 and comparative examples 1-7
Table 2 stability test results of the probiotic formulations prepared in examples 1 to 2 and comparative examples 1 to 7
TABLE 3 safety test results of probiotic formulations prepared in examples 1-2 and comparative examples 1-7
As can be seen from tables 1 to 3, the probiotic preparation is prepared by mixing pineapple peel fermentation liquor and soapberry fermentation liquor, adding brown sugar for secondary fermentation, and controlling conditions such as fermentation temperature, pH value, time and the like, so as to successfully extract active ingredients in a fermentation product and generate new substances.
From the comparative examples and comparative examples, it can be found that example 1 performs best in terms of probiotic bacteria count and activity, with higher stability and tolerance. In the temperature stability test, the activity of example 1 was reduced to a minimum extent, and the bacterial count was reduced to a minimum extent; in the humidity stability test, the activity decrease amplitude and the bacterial count decrease amplitude of the example 1 are smaller; in the acid and alkali resistance test, the activity decrease amplitude and the bacterial count decrease amplitude of the example 1 are smaller than those of other comparison groups. Furthermore, example 1 was less immunogenic and no drug resistance gene was detected.
In contrast, other comparative examples were lower in probiotic count and activity, and exhibited higher temperatures, humidity and poor acid-base stability. At the same time, these comparative examples were slightly increased in immunogenicity, and drug-resistant genes were detected.
In summary, example 1 is the optimal solution, with higher probiotic bacteria count and activity, and excellent stability and tolerability. Therefore, the embodiment 1 has better application prospect in the preparation of the probiotic preparation. The invention provides a more stable and more effective probiotic preparation through the fermentation preparation method of pineapple peel and soapberry, has the beneficial effects of improving the activity and stability of probiotics, providing a simple and efficient preparation method, providing a better and more reliable product and the like, and can provide better intestinal health guarantee for consumers.
The above examples are not intended to limit the scope of the invention nor the order of execution of the steps described. The present invention is obviously modified by a person skilled in the art in combination with the prior common general knowledge, and falls within the scope of protection defined by the claims of the present invention.
Claims (10)
1. A method for preparing a probiotic preparation with high activity and high stability, comprising the steps of:
(1) Mixing pineapple peel fermentation liquor and soapberry fermentation liquor together, adding brown sugar, and stirring to obtain a mixed solution;
(2) Filtering the mixed solution prepared in the step (1) to obtain filtrate, wherein the mesh size of a filter screen is 70 meshes during filtering;
(3) Pouring the filtrate obtained in the step (2) into a fermentation tank, adding saccharomycetes, stirring until the saccharomycetes are completely dissolved, and standing and fermenting for 36-63 h to obtain the probiotic preparation.
2. The preparation method according to claim 1, wherein in the step (1), the volume ratio of the pineapple peel fermentation liquid to the soapberry fermentation liquid is 1:3-3:1, and the mass ratio of the brown sugar to the mixed liquid of the pineapple peel fermentation liquid and the soapberry fermentation liquid is 1:3-1:5.
3. The method according to claim 1 or 2, wherein in step (1), the method for producing pineapple peel fermentation broth comprises the steps of:
a1. cleaning and peeling fresh pineapple peel, and cutting into small blocks with the thickness of 7mm multiplied by 7mm to 11mm multiplied by 11 mm;
b1. putting the pineapple peel small pieces in the step a1 into a fermentation tank according to a feed liquid ratio of 1: 4-1: 9, adding distilled water, and soaking for 35-55 min to obtain a solution to be fermented;
c1. adding saccharomycetes into the solution to be fermented in the step b1, standing and fermenting for 36-63 h, and filtering to obtain pineapple peel fermentation liquor.
4. The process according to claim 3, wherein in step c1, the yeast is added in an amount of 26 to 37% by volume of the solution to be fermented, the temperature is 20 to 29 ℃ during the stationary fermentation, the pH is 2.3 to 5.2, and the mesh size of the filter screen is 50 mesh during the filtration.
5. The preparation method according to claim 1, wherein in the step (1), the preparation method of the soapberry fermentation broth comprises the following steps:
a2. washing soapberry and grinding the soapberry into fine powder with the size of 95-137 meshes, putting the obtained soapberry powder into a fermentation tank, adding distilled water, and soaking the soapberry powder for 35-55 min to obtain a solution to be fermented;
b2. adding saccharomycetes into the solution to be fermented in the step a2, standing and fermenting for 36-63 h, and filtering to obtain soapberry fermentation liquor.
6. The method according to claim 5, wherein in step a2, the ratio of the soapberry powder to distilled water is 1: 4-1: 9.
7. the process according to claim 5, wherein in step b2, the yeast is added in an amount of 26 to 37% by volume of the solution to be fermented, the temperature is 20 to 29 ℃ during the stationary fermentation, the pH is 2.3 to 5.2, and the filtration is 50 mesh filtration.
8. The method according to claim 1, wherein in the step (3), the yeast is added in an amount of 28% -42% of the volume of the filtrate, and the temperature is 20 ℃ -29 ℃ and the pH value is 2.3-5.2 during the stationary fermentation.
9. A probiotic preparation prepared by the method according to any one of claims 1 to 8.
10. Use of the probiotic preparation according to claim 9 for preparing foods, medicines and health products, wherein the health products comprise health products with the functions of enhancing immunity, regulating intestinal flora, promoting digestion and relaxing bowels.
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