CN114081097B - Application of probiotic synergist in microbial fermentation - Google Patents

Application of probiotic synergist in microbial fermentation Download PDF

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CN114081097B
CN114081097B CN202210018936.1A CN202210018936A CN114081097B CN 114081097 B CN114081097 B CN 114081097B CN 202210018936 A CN202210018936 A CN 202210018936A CN 114081097 B CN114081097 B CN 114081097B
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probiotic
parts
synergist
extract
probiotics
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CN114081097A (en
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王文娟
杨艳
孙笑非
孙鸣
石磊
潘宝海
朱小龙
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Beijing Dorun Technology Co ltd
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
    • A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/116—Heterocyclic compounds
    • A23K20/121—Heterocyclic compounds containing oxygen or sulfur as hetero atom
    • 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
    • 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
    • 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)
    • A61K36/29—Berberidaceae (Barberry family), e.g. barberry, cohosh or mayapple
    • A61K36/296—Epimedium
    • 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)
    • A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
    • 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)
    • A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
    • A61K36/535—Perilla (beefsteak plant)
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The application relates to the technical field of microbial additives, and particularly discloses application of a probiotic synergist in microbial fermentation. The probiotic synergist is utilized for microbial fermentation to improve the maximum growth amount of microorganisms and shorten generation time; the probiotic synergist comprises one or more of fructus Perillae extract, herba Epimedii extract, and soybean flavone. The probiotic bacteria intestinal tract colonization and propagation method can promote colonization and propagation of probiotics in intestinal tracts.

Description

Application of probiotic synergist in microbial fermentation
Technical Field
The application relates to the technical field of microbial additives, in particular to application of a probiotic synergist in microbial fermentation.
Background
The balance of intestinal flora plays an important role in preventing and treating intestinal diseases. At present, in order to protect the microecological balance of intestinal tracts of domestic animals and poultry and restore normal flora of the intestinal tracts, a microecological preparation is provided, and a better effect is achieved in the animal husbandry. However, probiotics, which are foreign bacteria, are difficult to colonize in the intestinal tract, and in order to avoid the colonized probiotics from being eliminated, the probiotics need to be continuously taken in to maintain the effectiveness of the probiotics. Therefore, it is desirable to have probiotics that enter the intestinal tract propagate more rapidly and function as quickly as possible.
A probiotic synergist is a substance that selectively stimulates the growth of one or more bacteria in the host's intestinal tract, improving the health of the host without being digested by the host's gastrointestinal tract. The synbiotics, namely the product combining the probiotics and the probiotic synergist at the same time, selectively stimulates one or a limited number of probiotics to grow and metabolize in vivo by promoting exogenous live bacteria to colonize in the intestinal tract, thereby promoting the health of a host.
Based on the above-mentioned related technologies, the current research on the probiotic synergist is mainly limited to the effect of the probiotic synergist on bifidobacteria and lactobacilli subspecies, while the research on the probiotic synergist and other probiotics, especially on the study on the probiotic synergist and bacillus is less.
Disclosure of Invention
In order to promote the colonization and propagation of probiotics in intestinal tracts, the application provides application of microbial fermentation by using a probiotic synergist.
In a first aspect, the application provides an application of a probiotic synergist for microbial fermentation, which adopts the following technical scheme:
an application of probiotic synergist in microbial fermentation is provided, wherein the probiotic synergist is used for microbial fermentation to improve the maximum growth amount of microorganisms and shorten generation time; the probiotic synergist comprises one or more of fructus Perillae extract, herba Epimedii extract, and soybean flavone.
The probiotic synergist is used for microbial fermentation, so that the maximum growth amount of microorganisms is increased, and the generation time is shortened. Wherein the probiotic synergist comprises one or more of fructus Perillae extract, herba Epimedii extract, and soybean flavone. Through experimental analysis, the perilla seed extract, the epimedium extract and the daidzein can obviously improve the maximum growth amount of microorganisms and shorten the generation time of the microorganisms.
Perilla seed is the seed of plant Perilla frutescens, and has certain medicinal value, such as effects of descending qi, clearing phlegm, moistening lung, and relaxing intestine. Perilla seed contains a large amount of oil and fat and abundant protein. The perilla seed extract is an extract prepared from perilla seeds, so the perilla seed extract also contains a large amount of grease and protein, including linolenic acid, linoleic acid, oleic acid, lysine, methionine and the like. Therefore, the perilla seed extract can provide necessary nutrient substances for the growth and propagation of microorganisms, so that the growth, propagation and passage of the microorganisms are promoted, the growth amount of the microorganisms is increased, and the generation time is shortened.
The epimedium extract is epimedium (epimedium)Epimediumbrevicornum) The dried stems and leaves of the plants are used as raw materials and processed into the product. The epimedium extract has obvious inhibiting effect on the infection caused by staphylococcus aureus and poliovirus, and also has the effects of relieving cough, eliminating phlegm and relieving asthma. Therefore, the epimedium extract can obviously inhibit various viruses, thereby reducing the infection of harmful viruses and further ensuring the growth and reproduction of microorganisms, particularly probiotics.
The daidzein is mainly derived from legume of leguminous plant, and has high content in soybean. Daidzein mainly refers to 3-benzopyrone as the mother nucleus, and the total of the naturally occurring isoflavones in soybean is 12, and can be divided into 3 types, namely daidzin (Daidzingroups), genistin (genistinroups) and glycitinglops. Therefore, daidzein can be absorbed, metabolized and transformed by microorganisms, thereby promoting the growth and reproduction of microorganisms. Daidzein can also be converted by intestinal microorganisms into different products with higher and wider biological activity, thereby further promoting indirect utilization of daidzein by organisms.
Based on the above, the perilla seed extract, the epimedium extract and the daidzein can be used independently and randomly matched to be used as a probiotic synergist, and can be used for microbial fermentation to promote the propagation of microorganisms. And furthermore, the permanent planting and the propagation of microorganisms in the intestinal tract are promoted, so that the healthy development of intestinal microecology is further promoted, the barrier and absorption functions of the intestinal tract are further promoted, the utilization rate of the feed is improved, and the growth of animals is promoted. Particularly, the probiotic synergist can obviously promote the reproduction of probiotics and improve the colonization and reproduction of the probiotics in intestinal tracts.
In a particular embodiment, the probiotic synergist comprises perilla seed extract.
Preferably, the probiotic synergist comprises, by weight, 1000 parts of perilla seed extract, 0.068-1.32 parts.
In a specific embodiment, the perilla seed extract may be added in an amount of 0.068 parts, 0.340 parts, 0.670 parts, 1.320 parts.
In some specific embodiments, the perilla seed extract may be added in an amount of 0.068-0.340 parts, 0.068-0.670 parts, 0.068-1.320 parts, 0.340-0.670 parts, 0.340-1.320 parts, 0.670-1.320 parts.
In a particular embodiment, the probiotic synergist comprises epimedium extract.
Preferably, the probiotic synergist comprises 0.50-9.62 parts of epimedium extract by weight of 1000 parts.
In a specific embodiment, the epimedium extract may be added in an amount of 0.5 parts, 2.48 parts, 4.9 parts, 9.62 parts.
In some specific embodiments, the epimedium extract may be added in an amount of 0.5-2.48 parts, 0.5-4.9 parts, 0.5-9.62 parts, 2.48-4.9 parts, 2.48-9.62 parts, 4.9-9.62 parts.
In a particular embodiment, the probiotic synergist comprises icariin.
Preferably, the probiotic synergist comprises 0.50-9.62 parts of daidzein by weight of 1000 parts.
In a specific embodiment, the daidzein may be added in an amount of 0.5 parts, 2.48 parts, 4.9 parts, 9.62 parts.
In some specific embodiments, the amount of soy flavonoids added may be 0.5-2.48 parts, 0.5-4.9 parts, 0.5-9.62 parts, 2.48-4.9 parts, 2.48-9.62 parts, 4.9-9.62 parts.
In a specific embodiment, the probiotic synergist comprises perilla seed extract, epimedium extract.
In a particular embodiment, the probiotic synergist comprises perilla seed extract, soy flavonoids.
In a specific embodiment, the probiotic synergist comprises epimedium extract, soy flavonoids.
Preferably, the probiotic synergist is perilla seed extract, epimedium extract and soybean flavone.
Preferably, the probiotic synergist comprises the following components in 1000 parts by weight: 0.017-0.440 part of perilla seed extract; 0.280-2.420 parts of epimedium extract; 0.120-1.820 part of daidzein.
In a particular embodiment, the perilla seed extract may be 0.017 parts, 0.024 parts, 0.031 parts, 0.037 parts, 0.044 parts.
In some specific embodiments, the perilla seed extract may be 0.017 to 0.024 parts, 0.017 to 0.031 parts, 0.017 to 0.037 parts, 0.024 to 0.031 parts, 0.024 to 0.037 parts, 0.024 to 0.044 parts, 0.031 to 0.037 parts, 0.031 to 0.044 parts, 0.037 to 0.044 parts.
In a specific embodiment, the epimedium extract may be 0.280 parts, 0.850 parts, 1.350 parts, 1.890 parts, 2.420 parts.
In some specific embodiments, the epimedium extract may be 0280-0.850 parts, 0.280-1.350 parts, 0.280-1.890 parts, 0.850-1.350 parts, 0.850-1.890 parts, 0.850-2.420 parts, 1.350-1.890 parts, 1.350-2.420 parts, 1.890-2.420 parts.
In a particular embodiment, the daidzein can be 0.120 parts, 0.545 parts, 0.970 parts, 1.395 parts, 1.820 parts.
In some specific embodiments, the soy flavonoids may be 0.120-0.545, 0.120-0.970, 0.120-1.395, 0.545-0.970, 0.545-1.395, 0.545-1.820, 0.970-1.395, 0.970-1.820, 1.395-1.820.
By adopting the technical scheme, through experimental analysis, the perilla seed extract, the epimedium extract and the soybean flavone are cooperatively used as a probiotic synergist, so that the propagation of probiotics can be effectively promoted, and the colonization and propagation of the probiotics in the intestinal tract are further promoted, so that the healthy development of intestinal microecology is promoted, the barrier and absorption functions of the intestinal tract are promoted, the feed utilization rate is improved, and the growth of animals is promoted.
Preferably, the probiotic synergist promotes colonization and growth of said microorganisms in vivo.
Preferably, the probiotic synergist and the microorganism are used for preparing a microbial preparation.
Preferably, the microorganism is a probiotic.
Preferably, the probiotic is bacillus.
In a particular embodiment, the probiotic is bacillus subtilis.
Preferably, the microbial preparation further comprises a pharmacologically acceptable carrier and/or additive and/or feed material.
In a second aspect, the present application provides a probiotic synergist, which adopts the following technical scheme:
a probiotic synergist comprises one or more of fructus Perillae extract, herba Epimedii extract, and soybean flavone.
Preferably, the probiotic synergist comprises, by weight, 1000 parts of perilla seed extract, 0.068-1.32 parts.
Preferably, the probiotic synergist comprises 0.50-9.62 parts of epimedium extract by weight of 1000 parts.
Preferably, the probiotic synergistic agent comprises 0.50-9.62 parts of daidzein by weight of 1000 parts.
Preferably, the probiotic synergist comprises the following components in 1000 parts by weight: 0.017-0.440 part of perilla seed extract; 0.280-2.420 parts of epimedium extract; 0.120-1.820 part of daidzein.
In a third aspect, the present application provides a microbial preparation, which adopts the following technical scheme:
the microbial preparation comprises the probiotic synergist and probiotics, and the probiotics can be promoted to colonize and grow in vivo by the probiotic synergist.
Preferably, the probiotic is bacillus.
Preferably, the probiotic is bacillus subtilis.
Preferably, the microbial preparation further comprises a pharmacologically acceptable carrier and/or additive and/or feed material.
Preferably, the microbial preparation is a tablet, an ointment, a syrup, a capsule, a granule, a powder or an injection.
In a fourth aspect, the present application provides the use of a probiotic synergist as described above or a microbial preparation as described above for microbial fermentation. In summary, the present application has the following beneficial effects:
the perilla seed extract, the epimedium extract and the daidzein provided by the application can be used independently and used in any matching way to be used as a probiotic synergist to promote the propagation of probiotics. Further, the probiotic synergist provided by the application can promote the colonization and propagation of probiotics in the intestinal tract, so that the healthy development of intestinal microecology is further promoted, the barrier and absorption functions of the intestinal tract are further promoted, the feed utilization rate is improved, and the growth of animals is promoted.
Detailed Description
The present application provides a probiotic synergist for probiotics. The probiotic synergist comprises one or more of fructus Perillae extract, herba Epimedii extract, and soybean flavone.
Specifically, the probiotic synergist comprises, by weight, 1000 parts of perilla seed extract, 0.068-1.32 parts.
Specifically, in the probiotic synergist, the addition amount of the epimedium extract is 0.50-9.62 parts by weight based on 1000 parts by weight.
Specifically, the probiotic synergist comprises, by weight, 1000 parts of daidzein, 0.50-9.62 parts.
Further, the probiotic synergist comprises the following components in parts by weight according to 1000 parts by weight: 0.017-0.440 part of perilla seed extract; 0.280-2.420 parts of epimedium extract; 0.120-1.820 parts of daidzein.
The application also provides a microbial preparation. The microbial preparation comprises the probiotic synergist and probiotics, and the probiotics can be promoted to colonize and grow in vivo by the probiotic synergist. Specifically, the probiotics is bacillus. Further, the microbial preparation further comprises a pharmacologically acceptable carrier and/or additive and/or feed raw material. The microbial preparation can be tablets, ointments, syrups, capsules, granules, powders or injections.
The probiotic synergist or the microbial preparation provided by the application can be used for preparing medicines or foods for preventing and/or treating intestinal diseases.
The present application is described in further detail below with reference to examples 1-25, comparative examples 1-16, and the test tests.
Examples
Examples 1 to 12
Examples 1-12 each provide a probiotic synergist for a probiotic.
The preparation method of the probiotic synergist specifically comprises the following steps:
filtering and sterilizing the components for preparing the probiotic synergist, adding the components into water (rho =1000g/L) according to the addition amount of the components, fully dissolving, and fixing the volume to 1L by using water to prepare the probiotic synergist, and storing the probiotic synergist in a refrigerator at 4 ℃ for later use.
The components for preparing the probiotic synergist and the addition amount of each component are shown in table 1.
Table 1 the components and proportions of the probiotic synergists provided in examples 1-12 of table 1
Figure 26776DEST_PATH_IMAGE001
Comparative example
Comparative examples 1 to 16
Comparative examples 1-16 each provide a probiotic synergist for probiotics.
Comparative examples 1 to 4 differ from example 1 in that: the probiotic synergist was lactulose and the amount of lactulose added, as shown in table 2.
Comparative examples 5 to 8 differ from example 1 in that: the probiotic synergist is the addition amount of L-aspartic acid in the formula of L-aspartic acid, and is specifically shown in Table 2.
Comparative examples 9 to 12 differ from example 1 in that: the probiotic synergist was raffinose and the amount of raffinose added, as shown in table 2.
Comparative examples 13 to 16 differ from example 1 in that: the probiotic synergist was melibiose and the amount of melibiose added, as shown in table 2.
Table 2 components and proportions of the probiotic synergists provided in comparative examples 1-16
Figure 68550DEST_PATH_IMAGE002
Test for detection
(I) Probiotics
The influence of the probiotic synergist provided in examples 1-12 and comparative examples 1-16 on the growth of probiotics was examined by using Bacillus subtilis (CGMCC No. 1007) as the probiotic.
(II) Standard Curve for growth of Probiotics
1. Culture medium
(1) Peptone water medium: 10g of peptone and 5g of sodium chloride, and adding water to 1000 mL; adjusting pH to 7.0-7.2, and sterilizing at 121 deg.C for 30 min.
(2) Peptone solid medium: on the basis of the peptone water culture medium formula, 10g of nutrient agar is added.
2. Preparation method of standard curve
Inoculating 5mL of Bacillus subtilis seed solution into 50mL of peptone water culture medium, placing in a water bath oscillator at 37 ℃ and 90rpm, and culturing for 10h to obtain stock solution. The stock solution was diluted at 0, 20%, 50%, 60%, 80%, and 100%, respectively, and colorimetric at a wavelength of 660nm, and the OD value was measured at each ratio. Meanwhile, the number of bacteria at each ratio was counted by plate counting. The above operation was repeated four times, and the average of the number of bacteria was taken.
And drawing a standard curve by taking the OD value as a horizontal coordinate and the number of bacteria as a vertical coordinate to obtain a regression equation of the standard curve. The regression equation for the standard curve is as follows:
Y=1.964×X－0.106；r2=0.992；
wherein X represents OD value and Y represents bacterial count.
(III) detection method
The probiotic synergist prepared in examples 1-12 and comparative examples 1-16 was used to replace water to prepare peptone water culture medium corresponding to each example and each comparative example. Respectively taking 5mL (OD) of bacillus subtilis seed liquid600= 1) was inoculated into 50mL of each peptone water medium described above, and placed in a water bath shaker at 37 ℃ and 90rpm, and cultured for 12h as a detection group.
In addition, 5mL of Bacillus subtilis seed solution was inoculated into 50mL of peptone water medium, placed in a water bath shaker at 37 ℃ and 90rpm, and cultured for 12h as a control group.
The OD values of the peptone water culture media at a wavelength of 660nm were measured at 6h, 8h, 10h and 12h, respectively. The number of bacteria was counted at 0h using plate counting. And calculating to obtain corresponding bacteria number according to the regression equation of the standard curve and the OD value obtained by detection. And calculating a Logistic proliferation curve by using a 3-point method to obtain the maximum growth (K) of the bacillus subtilis.
And (4) selecting any two points in the logarithmic growth phase of the bacillus subtilis on the Logistic hyperplasia curve, and calculating the generation time (G). The formula for calculating the time (G) is as follows:
G=LN（Nt）-LN（N0）/（Tt-T0）；
wherein LN (N)t）、LN（N0) The numbers of bacteria, LN (N), at two points selected during the logarithmic growth phase are shown respectivelyt）＞LN（N0）；Tt、T0Respectively representing the times, T, corresponding to two points selected in the logarithmic growth phaset＞T0。
(IV) the results of the detection
The results are shown in Table 3.
TABLE 3 test results of examples 1 to 12 and comparative examples 1 to 16
Figure 176183DEST_PATH_IMAGE003
In combination with Table 3, it can be seen from the results of the tests of comparative examples 1 to 12 that different concentrations of perilla seed extract, epimedium extract and daidzein have different degrees of influence on the maximum growth amount and generation time of Bacillus subtilis. The application controls the adding amounts of the perilla seed extract, the epimedium extract and the soybean flavone in a proper range, can effectively improve the maximum growth amount of the bacillus subtilis and shorten the generation time of the bacillus subtilis. Therefore, the perilla seed extract, the epimedium extract and the soybean flavone are used as the probiotic synergist, so that the reproductive capacity of probiotics can be effectively improved.
As is apparent from the results of the comparative examples 1 to 4, when the addition amount of the perilla seed extract was controlled to 0.068 to 1.32 parts (per 1000 parts by weight), the original maximum growth amount of Bacillus subtilis was increased from 1.829 to 1.958 to 2.502, and when the growth amount was decreased from 55.1 to 52.8 to 54.6. Especially, when the addition amount of the perilla seed extract is controlled to be 0.340-0.670 parts (per 1000 parts by weight), the original maximum growth amount of the bacillus subtilis can be increased to 2.111-2.502, and the original maximum growth amount of the bacillus subtilis can be reduced to 52.8-54.0 in generation.
As is apparent from the results of the comparative examples 5 to 8, when the amount of the epimedium extract added is controlled to 0.5 to 9.62 parts per 1000 parts by weight, the original maximum growth amount of Bacillus subtilis can be increased from 1.829 to 2.172 to 2.617, and the original maximum growth amount of Bacillus subtilis can be decreased from 55.1 to 51.1 to 54.3. Especially, when the addition amount of the perilla seed extract is controlled to 2.48 to 4.9 parts (per 1000 parts by weight), the original maximum growth amount of the bacillus subtilis can be increased to 2.365 to 2.617, and the original maximum growth amount of the bacillus subtilis can be reduced to 51.1 to 53.5 in generation.
As is apparent from the results of the comparative examples 9 to 12, when the amount of the epimedium extract added is controlled to 0.5 to 9.62 parts per 1000 parts by weight, the original maximum growth amount of Bacillus subtilis can be increased from 1.829 to 2.203 to 2.552, and the original maximum growth amount of Bacillus subtilis can be decreased from 55.1 to 51.8 to 54.1. Especially, when the addition amount of the perilla seed extract is controlled to be 2.48 to 4.9 parts (per 1000 parts by weight), the original maximum growth amount of the bacillus subtilis can be increased to 2.289 to 2.552, and the original maximum growth amount of the bacillus subtilis can be reduced to 51.8 to 53.6 in generation.
In combination with Table 3, it can be seen by comparing the test results of comparative examples 1-16 that the maximum growth of Bacillus subtilis cannot be significantly increased when lactulose, L-aspartic acid, raffinose and Astragalus polysaccharides of different concentrations cannot effectively shorten the generation of Bacillus subtilis. Therefore, lactulose, L-aspartic acid, raffinose and astragalus polysaccharide have no significant effect on the growth of the bacillus subtilis.
In summary, the perilla seed extract, the epimedium extract and the daidzein provided in the embodiments 1 to 12 of the present application are all suitable for being used as probiotic synergists of probiotics, and respective addition amounts are provided, so that the propagation of the probiotics is promoted, and the determination and propagation capacities of the probiotics in intestinal tracts are further improved. While lactulose, L-aspartic acid, raffinose and astragalus polysaccharide given in comparative examples 1-16 are not suitable as probiotic synergists for probiotics, and cannot promote the propagation of probiotics within the test range of the application.
Examples 13 to 25
Examples 13-25 provide a probiotic synergist for probiotics, respectively.
The above embodiments are different from embodiment 1 in that: the probiotic synergist comprises fructus Perillae extract, herba Epimedii extract and daidzein, and is shown in Table 4.
Table 4 components and proportions of probiotic synergists provided in examples 13-25
Figure 633709DEST_PATH_IMAGE004
Test No. 2
(I) Probiotics
The influence of the probiotic synergist provided in examples 13-25 on the growth of probiotics was examined using Bacillus subtilis (accession number: CGMCC No. 1007) as the probiotic.
(II) Standard Curve for growth of Probiotics
See "detection test one".
(III) detection method
The subjects tested were probiotic synergists prepared in examples 13-25. The detection method is shown in detection test I.
(IV) the results of the detection
The results are shown in Table 5.
TABLE 5 examination results of examples 13 to 25
Figure 516258DEST_PATH_IMAGE005
Table 5 shows that the maximum growth of bacillus subtilis can be further significantly increased and the generation time of bacillus subtilis can be shortened by using the perilla seed extract, the epimedium extract and the daidzein as the probiotic synergist, as well as the detection results of comparative examples 13 to 25. Therefore, the application further selects a compound of perilla seed extract, epimedium extract and daidzein as a probiotic synergist so as to promote the propagation of probiotics. Still further, the application also provides a compounding ratio of the perilla seed extract, the epimedium extract and the soybean flavone, the compounding ratio of the perilla seed extract, the epimedium extract and the soybean flavone is controlled within the range of 0.017-0.440 part of the perilla seed extract, 0.280-2.420 parts of the epimedium extract and 0.120-1.820 parts of the soybean flavone (per 1000 parts by weight), the maximum growth amount of probiotics can be obviously improved, and meanwhile, the generation time of the probiotics is shortened, so that the reproductive capacity of the probiotics is effectively improved.
As can be seen from the results of the tests in comparative examples 13 to 17, when the perilla seed extract, the epimedium extract and the daidzein were used in combination, controlling the addition amount of the perilla seed extract to 0.017 to 0.44 parts (per 1000 parts by weight) increased the original maximum growth of Bacillus subtilis from 1.829 to 2.689 to 2.837 and decreased from 55.1 to 48.2 to 50.4 in generations. Especially, when the addition amount of the perilla seed extract is controlled to be 0.024-0.037 parts (per 1000 parts by weight), the original maximum growth amount of the bacillus subtilis can be increased to 2.776-2.837, and the original maximum growth amount of the bacillus subtilis can be reduced to 48.2-49.4 in generation.
As is clear from the results of comparing examples 15 and 19 to 21, when the perilla seed extract, the epimedium extract and the daidzein were used in combination, the original maximum growth of Bacillus subtilis was increased from 1.829 to 2.658-2.837 and from 55.1 to 48.2-50.6 at generations when the amount of the epimedium extract was controlled to 0.280-2.420 parts (per 1000 parts by weight). Especially, when the addition amount of the perilla seed extract is controlled to be 0.850-1.890 parts (per 1000 parts by weight), the original maximum growth amount of the bacillus subtilis can be increased to 2.754-2.837, and the original maximum growth amount of the bacillus subtilis can be reduced to 48.2-49.7 in generation.
From the results of the comparison of examples 15 and 19 to 21, it is found that when the perilla seed extract, the epimedium extract and the daidzein are compounded and used, the original maximum growth amount of the bacillus subtilis can be increased from 1.829 to 2.632-2.837 and the original maximum growth amount of the bacillus subtilis can be decreased from 55.1 to 48.2-50.6 when the addition amount of the daidzein is controlled to 0.120-1.820 parts (per 1000 parts by weight). Especially, when the addition amount of the perilla seed extract is controlled to 0.545-1.395 parts (per 1000 parts by weight), the original maximum growth amount of the bacillus subtilis can be increased to 2.748-2.837, and the generation time can be reduced to 48.2-49.4.
In summary, the compound of the perilla seed extract, the epimedium extract and the soy flavone provided in the embodiments 13 to 25 of the present application can be used as a probiotic synergist of probiotics, so as to further promote the propagation of the probiotics, and further improve the fixing and propagating abilities of the probiotics in intestinal tracts.
The present embodiment is only for explaining the present application, and it is not limited to the present application, and those skilled in the art can make modifications of the present embodiment without inventive contribution as needed after reading the present specification, but all of them are protected by patent law within the scope of the claims of the present application.

Claims (6)

1. The application of the probiotic synergist for microbial fermentation is characterized in that the probiotic synergist is used for microbial fermentation to improve the maximum growth amount of microorganisms and shorten the generation time; the probiotic synergist comprises the following components in parts by weight: 0.017-0.440 parts of perilla seed extract; 0.280-2.420 parts of epimedium extract; 0.120-1.820 parts of daidzein; the microorganism is bacillus subtilis.
2. The use of a probiotic synergist according to claim 1 for microbial fermentation, characterized in that said probiotic synergist comprises the following components in parts by weight, in 1000 parts by weight: 0.024-0.037 parts of perilla seed extract; 0.850-1.890 parts of epimedium extract; 0.545-1.395 parts of daidzein.
3. Use of a probiotic synergist according to claim 1 for microbial fermentation, characterized in that a microbial preparation is prepared using the probiotic synergist and the microorganism.
4. Use of a probiotic synergist for microbial fermentation according to claim 3, characterized in that said microbial preparation further comprises a pharmacologically acceptable carrier.
5. Use of a probiotic synergist according to claim 3 for microbial fermentation, characterized in that said microbial preparation also comprises pharmacologically acceptable additives.
6. Use of a probiotic synergist for microbial fermentation according to claim 3, characterized in that said microbial preparation also comprises pharmacologically acceptable feed raw materials.
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