CN102559765B - Yeast culture and application thereof in promoting proliferation of intestinal probiotics. - Google Patents
Yeast culture and application thereof in promoting proliferation of intestinal probiotics. Download PDFInfo
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- CN102559765B CN102559765B CN201210051962.0A CN201210051962A CN102559765B CN 102559765 B CN102559765 B CN 102559765B CN 201210051962 A CN201210051962 A CN 201210051962A CN 102559765 B CN102559765 B CN 102559765B
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Abstract
The invention discloses a yeast culture and application thereof in promoting proliferation of intestinal probiotics. The invention provides a method for preparing the yeast culture. The method comprises the following step of fermenting Saccharomyces cerevisiae with the collection number of CGMCC No. 2.1882 to obtain the yeast culture. The invention provides a process for preparing the yeast culture, by which the yeast culture with excellent performance is obtained. The yeast culture provided by the invention contains a variety of nutrients and can provide a nutrient substrate and nutritional active substances for growth of probiotics, thereby promoting the proliferation of the probiotics. The yeast culture and the application thereof in promoting the proliferation of the intestinal probiotics are of great value in the field of food or feed.
Description
Technical field
The present invention relates to a kind of yeast culture and the application in promotion proliferation of intestinal probiotics thereof.
Background technology
Along with the universal even abuse of feeding antibiotic, the negative effect such as resistance, drug residue that it brings becomes increasingly conspicuous.The restriction of feeding antibiotic is used and forbids is the inexorable trend of animal husbandry development, alternative promote intestinal beneficial bacterium propagation, improve intestinal environment, improve animal body immunizing power and develop, the novel green fodder additives that does not simultaneously decompose, do not produce resistance, noresidue for enteron aisle endoenzyme is the inevitable choice of sustainable development of animal husbandry.
Yeast culture (yeast culture, YC) refer to the micro-ecological goods that formed by yeast under specific process conditions after fully fermenting, mainly comprise substratum and a small amount of yeast cell of non-activity of yeast cell meta-bolites, variation after fermentation.As a kind of biologically active additives, yeast culture is nutritious, complicated component, contains VITAMIN, amino acid, digestive ferment, organic acid, polysaccharide and unidentified growth factor etc.According to relevant report, yeast culture can effectively improve animal gastrointestinal tract flora, promotes the probiotics breedings such as milk-acid bacteria, CELLULOLYTIC BACTERIUM.
The quality of yeast culture is subject to the impact of the factors such as bacterial classification, fermentation manufacturing technique and detection means.Yeast, under different culture environment, as differing temps, humidity or acid or alkali environment especially substrate composition, can produce different tunnings, and this wherein comprises a large amount of UGFs.The substratum using while producing yeast culture and the stability of composition, concentration and the bioavailability thereof of zymotechnique control parameter on meta-bolites in its end product have vital impact.Even if using identical barms in the situation that, different culture media composition or different fermentative production control technique can cause meta-bolites composition and concentration in yeast culture product to occur obvious difference.
Summary of the invention
The object of this invention is to provide a kind of yeast culture and the application in promotion proliferation of intestinal probiotics thereof.
The invention provides a kind of method of preparing yeast culture, is CGMCC to be numbered to 2.1882 yeast saccharomyces cerevisiae (Saccharomyces cerevisiae) ferment, and obtains yeast culture.
Described fermentation can comprise the steps:
(1) described yeast saccharomyces cerevisiae is seeded to seed culture medium, 18-40h (as 18-24h or 24-40h) is cultivated in 20-50 DEG C (as 20-28 DEG C or 28-50 DEG C), 100-200rpm (as 100-150rpm or 150-200rpm) jolting, obtains seed liquor;
(2) seed liquor of step (1) is seeded to fermention medium, 20-50 DEG C (as 20-30 DEG C or 30-50 DEG C) leaves standstill and cultivates 30-60h (30-45h or 45-60h).
The preparation method of described fermention medium is as follows: get 20-200g carbon source, 5-300g nitrogenous source, 0.1-10g (as 0.1-1g or 1-10g) KH
2pO
4, 0.1-10g (as 0.1-1g or 1-10g) MgSO
47H
2o, 0.1-4g (as 0.1-1g or 1-4g) MnSO
4h
2o, 0.1-4g (as 0.1-1g or 1-4g) CaCl
2, 0.1-4g (as 0.1-1g or 1-4g) CuSO
45H
2o, 0.1-4g (as 0.1-1g or 1-4g) ZnSO
47H
2o and 0.1-4g (as 0.1-1g or 1-4g) FeSO
44H
2o, water (as distilled water) dissolves and is settled to 1L.
The carbohydrate that described 20-200g carbon source specifically can contain sucrose by 10-100g glucose (as 10-50g or 50-100g) and 10-100g (as 10-50g or 50-100g) forms.Described 20-200g carbon source specifically can be the carbohydrate that 20-200g (as 20-100g or 100-200g) glucose or 20-200g (as 20-100g or 100-200g) contain sucrose.The described carbohydrate that contains sucrose is specially sucrose or molasses.
Described 5-300g nitrogenous source specifically can be by 2-100g (as 2-50g or 50-100g) yeast extract paste, 2-100g (as 2-50g or 50-100g) peptone and 1-100g (as 1-50g or 50-100g) ammonium sulfate composition, also can be formed by 2.5-150g (as 2.5-70g or 70-150g) yeast extract paste and 2.5-150g (as 2.5-70g or 70-150g) peptone, also can be formed by 2.5-150g (as 2.5-70g or 70-150g) yeast extract paste and 2.5-150g (as 2.5-70g or 70-150g) ammonium sulfate, also can be formed by 2.5-150g (as 2.5-70g or 70-150g) peptone and 2.5-150g (as 2.5-70g or 70-150g) ammonium sulfate.Described 5-300g nitrogenous source specifically can be 5-300g yeast extract paste or 5-300g peptone or 5-300g ammonium sulfate.
The preparation method of described seed culture medium is specific as follows: get 10-40g (as 10-20g or 20-40g) glucose, 5-40g (as 5-10g or 10-40g) yeast powder, 10-40g (as 10-20g or 20-40g) peptone and 0.1-4g (as 0.1-2g or 2-4g) MnSO
4h
2o, water (as distilled water) dissolves and is settled to 1L.
The pH value of described seed culture medium specifically can be nature pH.
The inoculum size of described seed liquor is 1%-30% (as 1%-15% or 15%-30%).The special seed liquor of inoculum size in this patent and the volume ratio of fermention medium.
Described seed liquor is seeded to the initial time of described fermention medium, and the concentration of described yeast saccharomyces cerevisiae is 1 × 10
5-3 × 10
6cfu/ml is (as 1 × 10
5-1.5 × 10
6cfu/ml or 1.5 × 10
6-3 × 10
6cfu/ml).
Described method also comprises the steps: to complete the centrifugal 5min of total system 4000rpm of fermentation, collects supernatant liquor.
Described method also comprises described fermentation biofilter (step that 0.22 μ m) filters for supernatant.
The yeast culture that arbitrary described method prepares above all belongs to protection scope of the present invention.
Described yeast culture can be used for promoting proliferation of intestinal probiotics.Described beneficial bacteria of intestinal tract specifically can be animal bifidobacteria, Lactobacterium acidophilum, plant lactobacillus, thermophilus streptococcus or lactobacillus delbruockii subspecies bulgaricus.
Described yeast culture can be used for the preparation of preparation promotion proliferation of intestinal probiotics.Described beneficial bacteria of intestinal tract specifically can be animal bifidobacteria, Lactobacterium acidophilum, plant lactobacillus, thermophilus streptococcus or lactobacillus delbruockii subspecies bulgaricus.
The present invention also protects a kind of method of cultivating beneficial bacteria of intestinal tract, is adding beneficial bacteria of intestinal tract (probiotic bacterium seed liquor) and described yeast culture for the substratum of cultivating described beneficial bacteria of intestinal tract, obtaining initial incubation system, then cultivating.Described beneficial bacteria of intestinal tract specifically can be animal bifidobacteria, Lactobacterium acidophilum, plant lactobacillus, thermophilus streptococcus or lactobacillus delbruockii subspecies bulgaricus.In described initial incubation system, the add-on of described yeast culture can be 0.1%-20% volume ratio (as 0.1%-10% volume ratio or 10%-20% volume ratio).In described initial incubation system, the add-on of described probiotics bacterial liquid can be 0.1%-20% volume ratio (as 0.1%-10% volume ratio or 10%-20% volume ratio).In described initial incubation system, described yeast culture and described probiotics bacterial liquid specifically can be the volume ratio of 1: 1.It is 10 that described probiotics bacterial liquid specifically can be concentration
7the bacterium liquid of cfu/ml.
Described animal bifidobacteria specifically can be CGMCC and is numbered 1.2268 bacterial strain.Described Lactobacterium acidophilum specifically can be CGMCC and is numbered 1.1878 bacterial strain.Described plant lactobacillus specifically can be CGMCC and is numbered 1.557 bacterial strain.Described thermophilus streptococcus specifically can be CGMCC and is numbered 1.2471 bacterial strain.Described lactobacillus delbruockii subspecies bulgaricus specifically can be CGMCC and is numbered 1.1480 bacterial strain.
The invention provides a kind of preparation technology of yeast culture, and obtained the yeast culture of excellent property.In yeast culture provided by the invention, contain various nutrients, can provide nutrition substrate and nutrient active matter for beneficial bacteria of intestinal tract growth, thereby promote proliferation of intestinal probiotics.The present invention has great value for field of food or field of fodder.
Embodiment
Following embodiment is convenient to understand better the present invention, but does not limit the present invention.Experimental technique in following embodiment, if no special instructions, is ordinary method.Test materials used in following embodiment, if no special instructions, is and purchases available from routine biochemistry reagent shop.Quantitative test in following examples, all arranges and repeats experiment, results averaged for three times.In embodiment, all adopt HCl or NaOH to regulate the pH value of substratum.
Substratum used in embodiment is as follows:
MRS substratum: purchased from Qingdao Hai Bo Bioisystech Co., Ltd, catalog number: HB0384-1.
PYG substratum: purchased from Qingdao Hai Bo Bioisystech Co., Ltd, catalog number: HB0398.
Glucose-yeast extract medium (pH 7.0): take 1.0g glucose, 5.0g peptone, 2.5g yeast extract paste, dissolve and be settled to 1L with distilled water.
Bacterial strain used in embodiment is as follows:
Yeast saccharomyces cerevisiae (Saccharomyces cerevisiae): CGMCC is numbered 2.1882.
Animal bifidobacteria (Bifidobacterium animalis is called for short BA): CGMCC is numbered 1.2268.
Lactobacterium acidophilum (Lactobacillus acidophilus is called for short LA): CGMCC is numbered 1.1878.
Plant lactobacillus (Lactobacillus plantarum is called for short LP): CGMCC is numbered 1.557.
Thermophilus streptococcus (Streptococcus thermophilus): CGMCC is numbered 1.2471.
Lactobacillus delbruockii subspecies bulgaricus (Lactobacillus delbrueckii subsp.bulgaricus): CGMCC is numbered 1.1480.
The preparation of embodiment 1, yeast culture and application
One, the preparation of substratum
Seed culture medium (natural pH): take 10g glucose, 5g yeast powder, 40g peptone and 4g MnSO
4h
2o, dissolves and is settled to 1L with distilled water; 121 DEG C, 0.1Mpa high-temperature sterilization 15min, for subsequent use.
Fermention medium (pH4.0): take carbon source (10g glucose and 10g sucrose), nitrogenous source (2g yeast extract paste, 2g peptone and 1g ammonium sulfate), 10g KH
2pO
4, 10g MgSO
47H
2o, 4g MnSO
4h
2o, 4g CaCl
2, 4g CuSO
45H
2o, 4g ZnSO
47H
2o and 4g FeSO
44H
2o, dissolves and is settled to 1L with distilled water; 121 DEG C, 0.1Mpa high-temperature sterilization 15min, for subsequent use.
Two, the preparation of yeast culture
1, the yeast saccharomyces cerevisiae after activation is accessed in seed culture medium, 28 DEG C, 150rpm (shimmy amplitude 25mm) shaking culture 24h, obtain seed liquor.
2, in 250ml triangular flask, 2ml seed liquor is seeded to 200ml fermention medium (being that inoculum size is 1%), the initial system that obtains fermenting, in the initial system of fermenting, the concentration of yeast saccharomyces cerevisiae is 1 × 10
5cfu/ml; 20 DEG C of standing for fermentation 60h of the initial system of fermenting, termination system obtains fermenting.
3, the centrifugal 5min of termination system 4000rpm (centrifugal radius is 13.5cm) that will ferment, collect supernatant liquor, by biofilter for supernatant liquor, (0.22 μ m) filters, and obtains cell-free filtrate (through aseptic the growing of flat board inspection), claims again aseptic yeast culture.
Three, the application of yeast culture
Five kinds of beneficial bacteria of intestinal tract (animal bifidobacteria, Lactobacterium acidophilum, plant lactobacillus, thermophilus streptococcus and lactobacillus delbruockii subspecies bulgaricus) are tested respectively as follows:
1, the beneficial bacteria of intestinal tract after activation is resuspended with sterilized water, obtaining bacteria concentration is 10
7the bacteria suspension of cfu/ml.
2, packet transaction
Experimental group: (Lactobacterium acidophilum, plant lactobacillus and lactobacillus delbruockii subspecies bulgaricus all adopt MRS substratum to add aseptic culture medium in centrifuge tube, animal bifidobacteria adopts PYG substratum, thermophilus streptococcus adopts glucose-yeast extract medium), the bacteria suspension that obtains of the aseptic yeast culture that obtains of step 2 and step 1, be initial system (liquid amount of centrifuge tube is 50%); The volumn concentration of aseptic yeast culture in initial system is 0.1%, and the volumn concentration of bacteria suspension in initial system is 0.1%.
Control group: sterilized water is replaced to the aseptic yeast culture in experimental group, other same experimental group.
Build the stopper of centrifuge tube in each group, cultivate 24h for 37 DEG C.
3, cultivate and finish rear employing ultraviolet-visible pectrophotometer and measure OD600 in total system (characterizing the biomass of beneficial bacteria of intestinal tract), the results are shown in Table 1.
The OD600 result of the experimental group in table 1 embodiment 1 and control group
| The OD600 result of experimental group | The OD600 result of control group | |
| Animal bifidobacteria | 0.648 (result after diluting 4 times) | 0.259 (result after diluting 4 times) |
| Lactobacterium acidophilum | 0.666 (result after diluting 11 times) | 0.238 (result after diluting 11 times) |
| Plant lactobacillus | 0.513 (result after diluting 11 times) | 0.223 (result after diluting 11 times) |
| Thermophilus streptococcus | 0.535 (result after diluting 11 times) | 0.243 (result after diluting 11 times) |
| Lactobacillus delbruockii subspecies bulgaricus | 0.487 (result after diluting 11 times) | 0.256 (result after diluting 11 times) |
Compared with control group, add after aseptic yeast culture, the OD600 of animal bifidobacteria has improved 1.5 times, the OD600 of Lactobacterium acidophilum and has improved 1.8 times, the OD600 of plant lactobacillus and improved 1.3 times, the OD600 of lactobacillus delbruockii subspecies bulgaricus and improved 0.9 times, the OD600 of thermophilus streptococcus and improved 1.2 times.
The preparation of embodiment 2, yeast culture and application
One, the preparation of substratum
Seed culture medium (natural pH): take 40g glucose, 40g yeast powder, 10g peptone and 0.1gMnSO
4h
2o, is settled to 1L after dissolving with distilled water; 121 DEG C, 0.1Mpa high-temperature sterilization 15min, for subsequent use.
Fermention medium (pH5.5): take carbon source (100g glucose and 100g sucrose), nitrogenous source (100g yeast extract paste, 100g peptone and 100g ammonium sulfate), 0.1g KH
2pO
4, 0.1g MgSO
47H
2o, 0.1g MnSO
4h
2o, 0.1g CaCl
2, 0.1g CuSO
45H
2o, 0.1g ZnSO
47H
2o and 0.1g FeSO
44H
2o, dissolves and is settled to 1L with distilled water; 121 DEG C, 0.1Mpa high-temperature sterilization 15min, for subsequent use.
Two, the preparation of yeast culture
1, the yeast saccharomyces cerevisiae after activation is accessed in seed culture medium, 20 DEG C, 100rpm (shimmy amplitude 25mm) shaking culture 18h, obtain seed liquor.
2, in 250ml triangular flask, 60ml seed liquor is seeded to 200ml fermention medium (being that inoculum size is 30%), the initial system that obtains fermenting, in the initial system of fermenting, the concentration of yeast saccharomyces cerevisiae is 3 × 10
6cfu/ml; 50 DEG C of standing for fermentation 30h of the initial system of fermenting, termination system obtains fermenting.
3, the centrifugal 5min of termination system 4000rpm (centrifugal radius 13.5cm) that will ferment, collect supernatant liquor, by biofilter for supernatant liquor, (0.22 μ m) filters, and obtains cell-free filtrate (through aseptic the growing of flat board inspection), claims again aseptic yeast culture.
Three, the application of yeast culture
Five kinds of beneficial bacteria of intestinal tract (animal bifidobacteria, Lactobacterium acidophilum, plant lactobacillus, thermophilus streptococcus and lactobacillus delbruockii subspecies bulgaricus) are tested respectively as follows:
1, with 1 of the step 3 of embodiment 1.
2, packet transaction
Experimental group: (Lactobacterium acidophilum, plant lactobacillus and lactobacillus delbruockii subspecies bulgaricus all adopt MRS substratum to add aseptic culture medium in centrifuge tube, animal bifidobacteria adopts PYG substratum, thermophilus streptococcus adopts glucose-yeast extract medium) in, the bacteria suspension that obtains of the aseptic yeast culture that obtains of step 2 and step 1, be initial system (liquid amount of centrifuge tube is 50%); The volumn concentration of aseptic yeast culture in initial system is 20%, and the volumn concentration of bacteria suspension in initial system is 20%.
Control group: fermention medium is replaced to the aseptic yeast culture in experimental group, other same experimental group.
Build the stopper of centrifuge tube in each group, cultivate 24h for 37 DEG C.
3, cultivate and finish rear employing ultraviolet-visible pectrophotometer and measure OD600 in total system (characterizing the biomass of beneficial bacteria of intestinal tract), the results are shown in Table 2.
The OD600 result of the experimental group in table 2 embodiment 2 and control group
| The OD600 result of experimental group | The OD600 result of control group | |
| Animal bifidobacteria | 0.803 (result after diluting 4 times) | 0.259 (result after diluting 4 times) |
| Lactobacterium acidophilum | 0.690 (result after diluting 11 times) | 0.238 (result after diluting 11 times) |
| Plant lactobacillus | 0.580 (result after diluting 11 times) | 0.223 (result after diluting 11 times) |
| Thermophilus streptococcus | 0.753 (result after diluting 11 times) | 0.243 (result after diluting 11 times) |
| Lactobacillus delbruockii subspecies bulgaricus | 0.640 (result after diluting 11 times) | 0.256 (result after diluting 11 times) |
Compared with control group, add after aseptic yeast culture, the OD600 of animal bifidobacteria has improved 2.1 times, the OD600 of Lactobacterium acidophilum and has improved 1.9 times, the OD600 of plant lactobacillus and improved 1.6 times, the OD600 of lactobacillus delbruockii subspecies bulgaricus and improved 1.5 times, the OD600 of thermophilus streptococcus and improved 2.1 times.
The preparation of embodiment 3, yeast culture and application
One, the preparation of substratum
Seed culture medium (natural pH): take 20g glucose, 10g yeast powder, 20g peptone and 2g MnSO
4h
2o, dissolves and is settled to 1L with distilled water; 121 DEG C, 0.1Mpa high-temperature sterilization 15min, for subsequent use.
Fermention medium (pH6.5) takes carbon source (50g glucose and 50g sucrose), nitrogenous source (50g yeast extract paste, 50g peptone and 50g ammonium sulfate), 1g KH
2pO
4, 1g MgSO
47H
2o, 1g MnSO
4h
2o, 1g CaCl
2, 1g CuSO
45H
2o, 1g ZnSO
47H
2o and 1g FeSO
44H
2o, dissolves and is settled to 1L with distilled water; 121 DEG C, 0.1Mpa high-temperature sterilization 15min, for subsequent use.
Two, the preparation of yeast culture
1, the yeast saccharomyces cerevisiae after activation is accessed in seed culture medium, 50 DEG C, 200rpm (shimmy amplitude 25mm) shaking culture 40h, obtain seed liquor.
2, in 250ml triangular flask, 30ml seed liquor is seeded to 200ml fermention medium (being that inoculum size is 15% volume ratio), the initial system that obtains fermenting, in the initial system of fermenting, the concentration of yeast saccharomyces cerevisiae is 1.5 × 10
6cfu/ml; 30 DEG C of standing for fermentation 45h of the initial system of fermenting, termination system obtains fermenting.
3, the centrifugal 5min of termination system 4000rpm (centrifugal radius 13.5cm) that will ferment, collect supernatant liquor, by biofilter for supernatant liquor, (0.22 μ m) filters, and obtains cell-free filtrate (through aseptic the growing of flat board inspection), claims again aseptic yeast culture.
Three, the application of yeast culture
Five kinds of beneficial bacteria of intestinal tract (animal bifidobacteria, Lactobacterium acidophilum, plant lactobacillus, thermophilus streptococcus and lactobacillus delbruockii subspecies bulgaricus) are tested respectively as follows:
1, with 1 of the step 3 of embodiment 1.
2, packet transaction
Experimental group: (Lactobacterium acidophilum, plant lactobacillus and lactobacillus delbruockii subspecies bulgaricus all adopt MRS substratum to add aseptic culture medium in centrifuge tube, animal bifidobacteria adopts PYG substratum, thermophilus streptococcus adopts glucose-yeast extract medium) in, the bacteria suspension that obtains of the aseptic yeast culture that obtains of step 2 and step 1, be initial system (liquid amount of centrifuge tube is 50%); The volumn concentration of aseptic yeast culture in initial system is 10%, and the volumn concentration of bacteria suspension in initial system is 10%.
Control group: fermention medium is replaced to the aseptic yeast culture in experimental group, other same experimental group.
The stopper of building centrifuge tube in each group, is placed in anaerobic jar, cultivates 24h for 37 DEG C.
3, cultivate and finish rear employing ultraviolet-visible pectrophotometer and measure OD600 in total system (characterizing the biomass of beneficial bacteria of intestinal tract), the results are shown in Table 3.
The OD600 result of the experimental group in table 3 embodiment 3 and control group
| The OD600 result of experimental group | The OD600 result of control group | |
| Animal bifidobacteria | 0.699 (result after diluting 4 times) | 0.259 (result after diluting 4 times) |
| Lactobacterium acidophilum | 0.595 (result after diluting 11 times) | 0.238 (result after diluting 11 times) |
| Plant lactobacillus | 0.513 (result after diluting 11 times) | 0.223 (result after diluting 11 times) |
| Thermophilus streptococcus | 0.632 (result after diluting 11 times) | 0.243 (result after diluting 11 times) |
| Lactobacillus delbruockii subspecies bulgaricus | 0.512 (result after diluting 11 times) | 0.256 (result after diluting 11 times) |
Compared with control group, add after aseptic yeast culture, the OD600 of animal bifidobacteria has improved 1.7 times, the OD600 of Lactobacterium acidophilum and has improved 1.5 times, the OD600 of plant lactobacillus and improved 1.3 times, the OD600 of lactobacillus delbruockii subspecies bulgaricus and improved 1.0 times, the OD600 of thermophilus streptococcus and improved 1.6 times.
The preparation of embodiment 4, yeast culture and application
One, the preparation of substratum
Seed culture medium (natural pH): with the seed culture medium of embodiment 3.
Fermention medium (pH6.5): carbon source is 20g glucose, nitrogenous source is made up of 2.5g yeast extract paste and 2.5g peptone, and other formula is with the fermention medium of embodiment 3.
Two, the preparation of yeast culture
With the step 2 of embodiment 3.
Three, the application of yeast culture
With the step 3 of embodiment 3.The results are shown in Table 4.
The OD600 result of the experimental group in table 4 embodiment 4 and control group
| The OD600 result of experimental group | The OD600 result of control group | |
| Animal bifidobacteria | 0.489 (result after diluting 4 times) | 0.259 (result after diluting 4 times) |
| Lactobacterium acidophilum | 0.415 (result after diluting 11 times) | 0.238 (result after diluting 11 times) |
| Plant lactobacillus | 0.332 (result after diluting 11 times) | 0.223 (result after diluting 11 times) |
| Thermophilus streptococcus | 0.461 (result after diluting 11 times) | 0.243 (result after diluting 11 times) |
| Lactobacillus delbruockii subspecies bulgaricus | 0.325 (result after diluting 11 times) | 0.256 (result after diluting 11 times) |
Compared with control group, add after aseptic yeast culture, the OD600 of animal bifidobacteria has improved 88.8%, the OD600 of Lactobacterium acidophilum has improved 74.4%, the OD600 of plant lactobacillus has improved 48.9%, the OD600 of lactobacillus delbruockii subspecies bulgaricus has improved 89.7%, the OD600 of thermophilus streptococcus has improved 27.0%.
The preparation of embodiment 5, yeast culture and application
One, the preparation of substratum
Seed culture medium (natural pH): with the seed culture medium of embodiment 3.
Fermention medium (pH6.5): carbon source is 200g glucose, nitrogenous source is made up of 150g yeast extract paste and 150g peptone, and other formula is with the fermention medium of embodiment 3.
Two, the preparation of yeast culture
With the step 2 of embodiment 3.
Three, the application of yeast culture
With the step 3 of embodiment 3.The results are shown in Table 5.
The OD600 result of the experimental group in table 5 embodiment 5 and control group
| The OD600 result of experimental group | The OD600 result of control group | |
| Animal bifidobacteria | 0.569 (result after diluting 4 times) | 0.259 (result after diluting 4 times) |
| Lactobacterium acidophilum | 0.471 (result after diluting 11 times) | 0.238 (result after diluting 11 times) |
| Plant lactobacillus | 0.403 (result after diluting 11 times) | 0.223 (result after diluting 11 times) |
| Thermophilus streptococcus | 0.528 (result after diluting 11 times) | 0.243 (result after diluting 11 times) |
| Lactobacillus delbruockii subspecies bulgaricus | 0.401 (result after diluting 11 times) | 0.256 (result after diluting 11 times) |
Compared with control group, add after aseptic yeast culture, the OD600 of animal bifidobacteria improved that 1.2 times, the OD600 of Lactobacterium acidophilum have improved 97.9%, the OD600 of plant lactobacillus has improved 80.7%, the OD600 of lactobacillus delbruockii subspecies bulgaricus has improved 56.6%, the OD600 of thermophilus streptococcus has improved 1.2 times.
The preparation of embodiment 6, yeast culture and application
One, the preparation of substratum
Seed culture medium (natural pH): with the seed culture medium of embodiment 3.
Fermention medium (pH6.5): carbon source is 100g glucose, nitrogenous source is made up of 70g yeast extract paste and 70g peptone, and other formula is with the fermention medium of embodiment 3.
Two, the preparation of yeast culture
With the step 2 of embodiment 3.
Three, the application of yeast culture
With the step 3 of embodiment 3.The results are shown in Table 6.
The OD600 result of the experimental group in table 6 embodiment 6 and control group
| The OD600 result of experimental group | The OD600 result of control group | |
| Animal bifidobacteria | 0.529 (result after diluting 4 times) | 0.259 (result after diluting 4 times) |
| Lactobacterium acidophilum | 0.425 (result after diluting 11 times) | 0.238 (result after diluting 11 times) |
| Plant lactobacillus | 0.453 (result after diluting 11 times) | 0.223 (result after diluting 11 times) |
| Thermophilus streptococcus | 0.522 (result after diluting 11 times) | 0.243 (result after diluting 11 times) |
| Lactobacillus delbruockii subspecies bulgaricus | 0.381 (result after diluting 11 times) | 0.256 (result after diluting 11 times) |
Compared with control group, add after aseptic yeast culture, the OD600 of animal bifidobacteria has improved 1.0 times, the OD600 of Lactobacterium acidophilum and has improved 78.6% times, the OD600 of plant lactobacillus and improved 1.0 times, the OD600 of lactobacillus delbruockii subspecies bulgaricus and improved 48.8% times, the OD600 of thermophilus streptococcus and improved 1.15 times.
The preparation of embodiment 7, yeast culture and application
One, the preparation of substratum
Seed culture medium (natural pH): with the seed culture medium of embodiment 3.
Fermention medium (pH6.5): carbon source is 20g sucrose, nitrogenous source is made up of 2.5g yeast extract paste and 2.5g ammonium sulfate, and other formula is with the fermention medium of embodiment 3.
Two, the preparation of yeast culture
With the step 2 of embodiment 3.
Three, the application of yeast culture
With the step 3 of embodiment 3.The results are shown in Table 7.
The OD600 result of the experimental group in table 7 embodiment 7 and control group
| The OD600 result of experimental group | The OD600 result of control group | |
| Animal bifidobacteria | 0.509 (result after diluting 4 times) | 0.259 (result after diluting 4 times) |
| Lactobacterium acidophilum | 0.415 (result after diluting 11 times) | 0.238 (result after diluting 11 times) |
| Plant lactobacillus | 0.320 (result after diluting 11 times) | 0.223 (result after diluting 11 times) |
| Thermophilus streptococcus | 0.441 (result after diluting 11 times) | 0.243 (result after diluting 11 times) |
| Lactobacillus delbruockii subspecies bulgaricus | 0.316 (result after diluting 11 times) | 0.256 (result after diluting 11 times) |
Compared with control group, add after aseptic yeast culture, the OD600 of animal bifidobacteria has improved 96.5%, the OD600 of Lactobacterium acidophilum has improved 74.4%, the OD600 of plant lactobacillus has improved 43.5%, the OD600 of lactobacillus delbruockii subspecies bulgaricus has improved 23.4%, the OD600 of thermophilus streptococcus has improved 81.5%.
The preparation of embodiment 8, yeast culture and application
One, the preparation of substratum
Seed culture medium (natural pH): with the seed culture medium of embodiment 3.
Fermention medium (pH6.5): carbon source is 200g sucrose, nitrogenous source is made up of 150g yeast extract paste and 150g ammonium sulfate, and other formula is with the fermention medium of embodiment 3.
Two, the preparation of yeast culture
With the step 2 of embodiment 3.
Three, the application of yeast culture
With the step 3 of embodiment 3.The results are shown in Table 8.
The OD600 result of the experimental group in table 8 embodiment 8 and control group
| The OD600 result of experimental group | The OD600 result of control group | |
| Animal bifidobacteria | 0.602 (result after diluting 4 times) | 0.259 (result after diluting 4 times) |
| Lactobacterium acidophilum | 0.489 (result after diluting 11 times) | 0.238 (result after diluting 11 times) |
| Plant lactobacillus | 0.399 (result after diluting 11 times) | 0.223 (result after diluting 11 times) |
| Thermophilus streptococcus | 0.540 (result after diluting 11 times) | 0.243 (result after diluting 11 times) |
| Lactobacillus delbruockii subspecies bulgaricus | 0.421 (result after diluting 11 times) | 0.256 (result after diluting 11 times) |
Compared with control group, add after aseptic yeast culture, the OD600 of animal bifidobacteria improved that 1.32 times, the OD600 of Lactobacterium acidophilum have improved that 1.05 times, the OD600 of plant lactobacillus have improved 78.9%, the OD600 of lactobacillus delbruockii subspecies bulgaricus has improved 64.5%, the OD600 of thermophilus streptococcus has improved 1.22 times.
The preparation of embodiment 9, yeast culture and application
One, the preparation of substratum
Seed culture medium (natural pH): with the seed culture medium of embodiment 3.
Fermention medium (pH6.5): carbon source is 100g sucrose, nitrogenous source is made up of 70g yeast extract paste and 70g ammonium sulfate, and other formula is with the fermention medium of embodiment 3.
Two, the preparation of yeast culture
With the step 2 of embodiment 3.
Three, the application of yeast culture
With the step 3 of embodiment 3.The results are shown in Table 9.
The OD600 result of the experimental group in table 9 embodiment 9 and control group
| The OD600 result of experimental group | The OD600 result of control group | |
| Animal bifidobacteria | 0.559 (result after diluting 4 times) | 0.259 (result after diluting 4 times) |
| Lactobacterium acidophilum | 0.465 (result after diluting 11 times) | 0.238 (result after diluting 11 times) |
| Plant lactobacillus | 0.373 (result after diluting 11 times) | 0.223 (result after diluting 11 times) |
| Thermophilus streptococcus | 0.482 (result after diluting 11 times) | 0.243 (result after diluting 11 times) |
| Lactobacillus delbruockii subspecies bulgaricus | 0.359 (result after diluting 11 times) | 0.256 (result after diluting 11 times) |
Compared with control group, add after aseptic yeast culture, the OD600 of animal bifidobacteria improved that 1.16 times, the OD600 of Lactobacterium acidophilum have improved 95.4%, the OD600 of plant lactobacillus has improved 67.3%, the OD600 of lactobacillus delbruockii subspecies bulgaricus has improved 40.2%, the OD600 of thermophilus streptococcus has improved 98.4%.
The preparation of embodiment 10, yeast culture and application
One, the preparation of substratum
Seed culture medium (natural pH): with the seed culture medium of embodiment 3.
Fermention medium (pH6.5): carbon source is 100g molasses, nitrogenous source is made up of 70g peptone and 70g ammonium sulfate, and other formula is with the fermention medium of embodiment 3.
Two, the preparation of yeast culture
With the step 2 of embodiment 3.
Three, the application of yeast culture
With the step 3 of embodiment 3.The results are shown in Table 10.
The OD600 result of the experimental group in table 10 embodiment 10 and control group
| The OD600 result of experimental group | The OD600 result of control group | |
| Animal bifidobacteria | 0.502 (result after diluting 4 times) | 0.259 (result after diluting 4 times) |
| Lactobacterium acidophilum | 0.410 (result after diluting 11 times) | 0.238 (result after diluting 11 times) |
| Plant lactobacillus | 0.322 (result after diluting 11 times) | 0.223 (result after diluting 11 times) |
| Thermophilus streptococcus | 0.441 (result after diluting 11 times) | 0.243 (result after diluting 11 times) |
| Lactobacillus delbruockii subspecies bulgaricus | 0.320 (result after diluting 11 times) | 0.256 (result after diluting 11 times) |
Compared with control group, add after aseptic yeast culture, the OD600 of animal bifidobacteria has improved 93.8%, the OD600 of Lactobacterium acidophilum has improved 72.3%, the OD600 of plant lactobacillus has improved 44.4%, the OD600 of lactobacillus delbruockii subspecies bulgaricus has improved 25% times, the OD600 of thermophilus streptococcus and improved 81.5%.
The preparation of embodiment 11, yeast culture and application
One, the preparation of substratum
Seed culture medium (natural pH): with the seed culture medium of embodiment 3.
Fermention medium (pH6.5): carbon source is 200g molasses, nitrogenous source is made up of 150g peptone and 150g ammonium sulfate, and other formula is with the fermention medium of embodiment 3.
Two, the preparation of yeast culture
With the step 2 of embodiment 3.
Three, the application of yeast culture
With the step 3 of embodiment 3.The results are shown in Table 11.
The OD600 result of the experimental group in table 11 embodiment 11 and control group
| The OD600 result of experimental group | The OD600 result of control group |
| Animal bifidobacteria | 0.567 (result after diluting 4 times) | 0.259 (result after diluting 4 times) |
| Lactobacterium acidophilum | 0.465 (result after diluting 11 times) | 0.238 (result after diluting 11 times) |
| Plant lactobacillus | 0.470 (result after diluting 11 times) | 0.223 (result after diluting 11 times) |
| Thermophilus streptococcus | 0.504 (result after diluting 11 times) | 0.243 (result after diluting 11 times) |
| Lactobacillus delbruockii subspecies bulgaricus | 0.392 (result after diluting 11 times) | 0.256 (result after diluting 11 times) |
Compared with control group, add after aseptic yeast culture, the OD600 of animal bifidobacteria improved that 1.2 times, the OD600 of Lactobacterium acidophilum have improved 95.4%, the OD600 of plant lactobacillus has improved that 1.1 times, the OD600 of lactobacillus delbruockii subspecies bulgaricus have improved 53.1%, the OD600 of thermophilus streptococcus has improved 1.07 times.
The preparation of embodiment 12, yeast culture and application
One, the preparation of substratum
Seed culture medium (natural pH): with the seed culture medium of embodiment 3.
Fermention medium (pH6.5): carbon source is 20g molasses, nitrogenous source is made up of 2.5g peptone and 2.5g ammonium sulfate, and other formula is with the fermention medium of embodiment 3.
Two, the preparation of yeast culture
With the step 2 of embodiment 3.
Three, the application of yeast culture
With the step 3 of embodiment 3.The results are shown in Table 12.
The OD600 result of the experimental group in table 12 embodiment 12 and control group
| The OD600 result of experimental group | The OD600 result of control group | |
| Animal bifidobacteria | 0.452 (result after diluting 4 times) | 0.259 (result after diluting 4 times) |
| Lactobacterium acidophilum | 0.366 (result after diluting 11 times) | 0.238 (result after diluting 11 times) |
| Plant lactobacillus | 0.302 (result after diluting 11 times) | 0.223 (result after diluting 11 times) |
| Thermophilus streptococcus | 0.401 (result after diluting 11 times) | 0.243 (result after diluting 11 times) |
| Lactobacillus delbruockii subspecies bulgaricus | 0.312 (result after diluting 11 times) | 0.256 (result after diluting 11 times) |
Compared with control group, add after aseptic yeast culture, the OD600 of animal bifidobacteria has improved 74.5%, the OD600 of Lactobacterium acidophilum has improved 53.8%, the OD600 of plant lactobacillus has improved 35.4%, the OD600 of lactobacillus delbruockii subspecies bulgaricus has improved 21.9%, the OD600 of thermophilus streptococcus has improved 65.0%.
The preparation of embodiment 13, yeast culture and application
One, the preparation of substratum
Seed culture medium (natural pH): with the seed culture medium of embodiment 3.
Fermention medium (pH6.5): carbon source is 20g glucose, nitrogenous source is 5g yeast extract paste, other formula is with the fermention medium of embodiment 3.
Two, the preparation of yeast culture
With the step 2 of embodiment 3.
Three, the application of yeast culture
With the step 3 of embodiment 3.
Three, the application of yeast culture
With the step 3 of embodiment 3.The results are shown in Table 13.
The OD600 result of the experimental group in table 13 embodiment 13 and control group
| The OD600 result of experimental group | The OD600 result of control group | |
| Animal bifidobacteria | 0.432 (result after diluting 4 times) | 0.259 (result after diluting 4 times) |
| Lactobacterium acidophilum | 0.342 (result after diluting 11 times) | 0.238 (result after diluting 11 times) |
| Plant lactobacillus | 0.285 (result after diluting 11 times) | 0.223 (result after diluting 11 times) |
| Thermophilus streptococcus | 0.375 (result after diluting 11 times) | 0.243 (result after diluting 11 times) |
| Lactobacillus delbruockii subspecies bulgaricus | 0.290 (result after diluting 11 times) | 0.256 (result after diluting 11 times) |
Compared with control group, add after aseptic yeast culture, the OD600 of animal bifidobacteria has improved 66.8%, the OD600 of Lactobacterium acidophilum has improved 43.7%, the OD600 of plant lactobacillus has improved 27.8%, the OD600 of lactobacillus delbruockii subspecies bulgaricus has improved 13.3%, the OD600 of thermophilus streptococcus has improved 54.3%.
The preparation of embodiment 14, yeast culture and application
One, the preparation of substratum
Seed culture medium (natural pH): with the seed culture medium of embodiment 3.
Fermention medium (pH6.5): carbon source is 200g glucose, nitrogenous source is 300g yeast extract paste, other formula is with the fermention medium of embodiment 3.
Two, the preparation of yeast culture
With the step 2 of embodiment 3.
Three, the application of yeast culture
With the step 3 of embodiment 3.
Three, the application of yeast culture
With the step 3 of embodiment 3.The results are shown in Table 14.
The OD600 result of the experimental group in table 14 embodiment 14 and control group
| The OD600 result of experimental group | The OD600 result of control group | |
| Animal bifidobacteria | 0.555 (result after diluting 4 times) | 0.259 (result after diluting 4 times) |
| Lactobacterium acidophilum | 0.490 (result after diluting 11 times) | 0.238 (result after diluting 11 times) |
| Plant lactobacillus | 0.421 (result after diluting 11 times) | 0.223 (result after diluting 11 times) |
| Thermophilus streptococcus | 0.410 (result after diluting 11 times) | 0.243 (result after diluting 11 times) |
| Lactobacillus delbruockii subspecies bulgaricus | 0.344 (result after diluting 11 times) | 0.256 (result after diluting 11 times) |
Compared with control group, add after aseptic yeast culture, the OD600 of animal bifidobacteria improved that 1.14 times, the OD600 of Lactobacterium acidophilum have improved that 1.06 times, the OD600 of plant lactobacillus have improved 88.8%, the OD600 of lactobacillus delbruockii subspecies bulgaricus has improved 34.4%, the OD600 of thermophilus streptococcus has improved 68.7%.
The preparation of embodiment 15, yeast culture and application
One, the preparation of substratum
Seed culture medium (natural pH): with the seed culture medium of embodiment 3.
Fermention medium (pH6.5): carbon source is 20g sucrose, nitrogenous source is 5g peptone, other formula is with the fermention medium of embodiment 3.
Two, the preparation of yeast culture
With the step 2 of embodiment 3.
Three, the application of yeast culture
With the step 3 of embodiment 3.
Three, the application of yeast culture
With the step 3 of embodiment 3.The results are shown in Table 15.
The OD600 result of the experimental group in table 15 embodiment 15 and control group
| The OD600 result of experimental group | The OD600 result of control group | |
| Animal bifidobacteria | 0.445 (result after diluting 4 times) | 0.259 (result after diluting 4 times) |
| Lactobacterium acidophilum | 0.358 (result after diluting 11 times) | 0.238 (result after diluting 11 times) |
| Plant lactobacillus | 0.289 (result after diluting 11 times) | 0.223 (result after diluting 11 times) |
| Thermophilus streptococcus | 0.390 (result after diluting 11 times) | 0.243 (result after diluting 11 times) |
| Lactobacillus delbruockii subspecies bulgaricus | 0.295 (result after diluting 11 times) | 0.256 (result after diluting 11 times) |
Compared with control group, add after aseptic yeast culture, the OD600 of animal bifidobacteria has improved 71.8%, the OD600 of Lactobacterium acidophilum has improved 50.4%, the OD600 of plant lactobacillus has improved 29.6%, the OD600 of lactobacillus delbruockii subspecies bulgaricus has improved 15.2%, the OD600 of thermophilus streptococcus has improved 60.5%.
The preparation of embodiment 16, yeast culture and application
One, the preparation of substratum
Seed culture medium (natural pH): with the seed culture medium of embodiment 3.
Fermention medium (pH6.5): carbon source is 200g sucrose, nitrogenous source is 300g peptone, other formula is with the fermention medium of embodiment 3.
Two, the preparation of yeast culture
With the step 2 of embodiment 3.
Three, the application of yeast culture
With the step 3 of embodiment 3.
Three, the application of yeast culture
With the step 3 of embodiment 3.The results are shown in Table 16.
The OD600 result of the experimental group in table 16 embodiment 16 and control group
| The OD600 result of experimental group | The OD600 result of control group | |
| Animal bifidobacteria | 0.535 (result after diluting 4 times) | 0.259 (result after diluting 4 times) |
| Lactobacterium acidophilum | 0.452 (result after diluting 11 times) | 0.238 (result after diluting 11 times) |
| Plant lactobacillus | 0.396 (result after diluting 11 times) | 0.223 (result after diluting 11 times) |
| Thermophilus streptococcus | 0.377 (result after diluting 11 times) | 0.243 (result after diluting 11 times) |
| Lactobacillus delbruockii subspecies bulgaricus | 0.310 (result after diluting 11 times) | 0.256 (result after diluting 11 times) |
Compared with control group, add after aseptic yeast culture, the OD600 of animal bifidobacteria improved that 1.07 times, the OD600 of Lactobacterium acidophilum have improved 89.9%, the OD600 of plant lactobacillus has improved 77.6%, the OD600 of lactobacillus delbruockii subspecies bulgaricus has improved 21.1%, the OD600 of thermophilus streptococcus has improved 55.1%.
The preparation of embodiment 17, yeast culture and application
One, the preparation of substratum
Seed culture medium (natural pH): with the seed culture medium of embodiment 3.
Fermention medium (pH6.5): carbon source is 20g molasses, nitrogenous source is 5g ammonium sulfate, other formula is with the fermention medium of embodiment 3.
Two, the preparation of yeast culture
With the step 2 of embodiment 3.
Three, the application of yeast culture
With the step 3 of embodiment 3.
Three, the application of yeast culture
With the step 3 of embodiment 3.The results are shown in Table 17.
The OD600 result of the experimental group in table 17 embodiment 17 and control group
| The OD600 result of experimental group | The OD600 result of control group | |
| Animal bifidobacteria | 0.405 (result after diluting 4 times) | 0.259 (result after diluting 4 times) |
| Lactobacterium acidophilum | 0.329 (result after diluting 11 times) | 0.238 (result after diluting 11 times) |
| Plant lactobacillus | 0.260 (result after diluting 11 times) | 0.223 (result after diluting 11 times) |
| Thermophilus streptococcus | 0.355 (result after diluting 11 times) | 0.243 (result after diluting 11 times) |
| Lactobacillus delbruockii subspecies bulgaricus | 0.269 (result after diluting 11 times) | 0.256 (result after diluting 11 times) |
Compared with control group, add after aseptic yeast culture, the OD600 of animal bifidobacteria has improved 56.4%, the OD600 of Lactobacterium acidophilum has improved 38.2%, the OD600 of plant lactobacillus has improved 16.6%, the OD600 of lactobacillus delbruockii subspecies bulgaricus has improved 5.1%, the OD600 of thermophilus streptococcus has improved 46.1%.
The preparation of embodiment 18, yeast culture and application
One, the preparation of substratum
Seed culture medium (natural pH): with the seed culture medium of embodiment 3.
Fermention medium (pH6.5): carbon source is 200g molasses, nitrogenous source is 300g ammonium sulfate, other formula is with the fermention medium of embodiment 3.
Two, the preparation of yeast culture
With the step 2 of embodiment 3.
Three, the application of yeast culture
With the step 3 of embodiment 3.
Three, the application of yeast culture
With the step 3 of embodiment 3.The results are shown in Table 18.
The OD600 result of the experimental group in table 18 embodiment 18 and control group
| The OD600 result of experimental group | The OD600 result of control group | |
| Animal bifidobacteria | 0.505 (result after diluting 4 times) | 0.259 (result after diluting 4 times) |
| Lactobacterium acidophilum | 0.421 (result after diluting 11 times) | 0.238 (result after diluting 11 times) |
| Plant lactobacillus | 0.360 (result after diluting 11 times) | 0.223 (result after diluting 11 times) |
| Thermophilus streptococcus | 0.352 (result after diluting 11 times) | 0.243 (result after diluting 11 times) |
| Lactobacillus delbruockii subspecies bulgaricus | 0.270 (result after diluting 11 times) | 0.256 (result after diluting 11 times) |
Compared with control group, add after aseptic yeast culture, the OD600 of animal bifidobacteria has improved 95.0%, the OD600 of Lactobacterium acidophilum has improved 76.9%, the OD600 of plant lactobacillus has improved 61.4%, the OD600 of lactobacillus delbruockii subspecies bulgaricus has improved 5.5%, the OD600 of thermophilus streptococcus has improved 44.9%.
Claims (4)
1. preparing a method for yeast culture, is CGMCC to be numbered to 2.1882 yeast saccharomyces cerevisiae (Saccharomyces cerevisiae) ferment, and obtains yeast culture;
Described fermentation comprises the steps:
(1) described yeast saccharomyces cerevisiae is seeded to seed culture medium, 20-50 DEG C, 100-200rpm jolting cultivation 18-40h, obtain seed liquor;
(2) seed liquor of step (1) is seeded to fermention medium, 20-50 DEG C leaves standstill cultivation 30-60h;
The preparation method of described fermention medium is as follows: get 20-200g carbon source, 5-300g nitrogenous source, 0.1-10g KH
2pO
4, 0.1-10g MgSO
47H
2o, 0.1-4g MnSO
4h
2o, 0.1-4g CaCl
2, 0.1-4g CuSO
45H
2o, 0.1-4gZnSO
47H
2o and 0.1-4g FeSO
44H
2o, by water dissolution and be settled to 1L;
Described carbon source is glucose and/or sucrose and/or molasses; Described nitrogenous source is yeast extract paste and/or peptone and/or ammonium sulfate;
The preparation method of described seed culture medium is specific as follows: get 10-40g glucose, 5-40g yeast powder, 10-40g peptone and 0.1-4g MnSO
4h
2o, by water dissolution and be settled to 1L;
When described seed liquor is seeded to described fermention medium, the volume proportion of described seed liquor and described fermention medium is 1-30:100.
2. the yeast culture that described in claim 1, method prepares.
3. the application of yeast culture in the preparation of preparation promotion proliferation of intestinal probiotics described in claim 2;
Described beneficial bacteria of intestinal tract is animal bifidobacteria, Lactobacterium acidophilum, plant lactobacillus, thermophilus streptococcus or lactobacillus delbruockii subspecies bulgaricus.
4. cultivating a method for beneficial bacteria of intestinal tract, is to add yeast culture described in described beneficial bacteria of intestinal tract and claim 2 for the substratum of cultivating beneficial bacteria of intestinal tract, obtaining initial incubation system, then cultivating;
Described beneficial bacteria of intestinal tract is animal bifidobacteria, Lactobacterium acidophilum, plant lactobacillus, thermophilus streptococcus or lactobacillus delbruockii subspecies bulgaricus.
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