CN109593690A - The chemical stress removal method of the not tally diplobacterium of High Density Cultivation - Google Patents
The chemical stress removal method of the not tally diplobacterium of High Density Cultivation Download PDFInfo
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Abstract
The present invention provides a kind of chemical stress removal methods of High Density Cultivation bifidobacterium bifidum, comprising the following steps: the culture medium of a. preparation bifidobacterium bifidum；B. antioxidant is added in the culture medium that step (a) is prepared, and carries out sterilization treatment；C. it is inoculated with；D. continue to add antioxidant in incubation；E. after cultivating, count plate is carried out using colony counting method.Antioxidant in the step (b) is sodium isoascorbate, and the antioxidant in the step (d) is sodium ascorbate.Chemical stress removal method provided by the invention is able to ascend the culture density of bifidobacterium bifidum, promotes the culture efficiency of probiotics, saves production space and cost.
Description
Technical field
The invention belongs to probiotic's culture technical fields, and in particular to the height density cultural method of bifidobacterium bifidum.
Background technique
Some bacteriums (such as lactobacillus and Bifidobacterium) have excellent functional characteristic, can provide one for the mankind
The health-care effect of series, thus it is referred to as probiotics by people.In the production process of probiotics, it is desirable to by improving benefit
The density of raw bacterium culture, Lai Jieyue production space, and obtain the formulation products of higher prebiotic bacterial content.But probiotics is in life
Certain metabolite can be generated in growth process, and there is the competitive relation to nutriment, excessively high culture each other
Density, by probiotics growth and breeding have adverse effect on.
Bifidobacterium bifidum strictly anaerobic cannot be proliferated, the residual of oxygen, meeting in fermentation liquid under conditions of being full of oxygen
Coercion is generated to bifidobacterium bifidum, is unfavorable for the proliferation of bifidobacterium bifidum.The present invention is the not tally bifid bar of probiotics
The High Density Cultivation of bacterium provides a new technical method, utilizes antioxidant -- and arabo-ascorbic acid and ascorbic acid eliminate training
The dissolved oxygen supported in base realizes the High Density Cultivation of bifidobacterium bifidum to eliminate dissolved oxygen to the coercion of bifidobacterium bifidum,
This method is known as chemical stress removal method.The chemical stress removal method is related to following reaction: ascorbic acid+oxygen → hydroascorbic acid+
Water；Arabo-ascorbic acid+oxygen → dehydrogenation arabo-ascorbic acid+water.
With the variation of socio-economic development and natural environment, rhythm of life is accelerated, and various pressure are also comed one after another, people
There is the problem of various healths aspects.And probiotics and people carry out the connection of health own profound.It is prebiotic
Bacterium is a kind of active microorganism beneficial to host, is to be colonized in human body intestinal canal, in reproductive system, can generate definite health efficacy
So as to improve the active beneficial microorganism general name of host's microecological balance, performance beneficial effect.Probiotics is roughly divided into three classes,
Including lactobacillus (such as lactobacillus acidophilus, Lactobacillus casei, Lactobacillus Jensenii, Raman lactobacillus), Bifidobacterium is (such as length
Bifidobacterium, bifidobacterium breve, oval Bifidobacterium, bifidobacterium thermophilum etc.), Gram positive cocci (such as streptococcus fecalis, cream
Coccus, intermediary streptococcus etc.), in addition, there are also the scopes that some yeast can also be included into probiotics.
With going deep into for probiotics and human health relationship research, it has been found that probiotics integrates with following several sides
The effect in face: (1) it improves immunity: generating a large amount of immunoglobulin, activate the immunocyte of body, so that external bacterium is difficult to
It retains in vivo, reaches immunoregulation effect；(2) improve gastrointestinal function: forming protective barrier in gut mucosal surface, resisted
Evil bacterium destroys the invasion of intestinal mucosa, causes the environment for being unfavorable for harmful bacteria growth, inhibits growth, the breeding of harmful bacteria；(3)
Trophism: fermentation generates lactic acid and acetic acid, can improve the utilization rate of calcium, phosphorus, iron, promotes the digestion and suction of related nutritional substance
It receives；(4) lead to intestines profit just: can produce a large amount of acidic materials in the intestine, stimulate intestines peristalsis, play the effect etc. of defaecation.Enteron aisle is micro-
The research and development of ecological agent product can carry out positive promote meaning to the health care belt of the mankind, with important economic value and
Community health's meaning.
Chinese patent announces CN108783462A and discloses a kind of industrial process of beneficial bacteria of intestinal tract preparation, including
(1) Lactobacillus casei, Lactobacillus rhamnosus, bifidobacterium bifidum, lactobacillus acidophilus, streptococcus thermophilus, Lactobacillus delbrueckii are protected
Leah subspecies etc. are added to carry out mass propgation respectively with fermentor, (2) obtain wet bacterium mud by centrifugation and sterile water washing, by wet bacterium
Mud is uniformly mixed with special protective preparation by 1:1, and (3) pelletized wet bacterium mud and protectant mixture, round as a ball, cryogenic vacuum
Dry, coating, is made beneficial bacteria of intestinal tract preparation.The invention provide condition of culture under, probiotics can high-density growth breeding,
It collects obtained wet bacterium mud to be not required to be freeze-dried, vacuum and low temperature rapid draing is directly carried out with a kind of special protective preparation and can keep
Thallus large number of viable saves the energy, reduces production cost.But this method needs to add a large amount of protection in use
Agent reduces the relative amount of viable count in probiotics preparation.Chinese patent announcement CN102614225A discloses a kind of compound
Probiotics viable bacteria preparation and its industrialized preparing process, the complex probiotics viable bacteria preparation are double by lactobacillus acidophilus A-10, youth
Discrimination bacillus B-13, auxiliary protection carrier mix, and wherein compound probiotic bacterium powder is 2.0%-30%, and auxiliary protection carrier is
70-98%, although the invention improves the number of viable and survival rate of probiotics to a certain extent, still exist auxiliary
The problem for helping protection carrier dosage big.
So being badly in need of a kind of industrial process for the probiotics that Viable detection is high, protective agent additive amount is few at present.
Summary of the invention
The present invention provides a kind of chemical stress removal methods of High Density Cultivation bifidobacterium bifidum, are able to ascend not tally bifid
The culture density of bacillus saves production space and cost.
The present invention provides a kind of chemical stress removal methods of High Density Cultivation bifidobacterium bifidum, and the method includes following
Step:
A. the culture medium of bifidobacterium bifidum is prepared；
B. antioxidant is added in the culture medium that step (a) is prepared, and carries out sterilization treatment；
C. it is inoculated with；
D. continue to add antioxidant in incubation；
E. after cultivating, count plate is carried out using colony counting method.
Culture medium in the step (a) are as follows: glucose 5.0-15.0g/L, pancreatin hydrolysis casein 2.0-15.0g/L,
Corn pulp 1.5-5.5g/L, sodium citrate 1.0-5.0g/L, dipotassium hydrogen phosphate 0.2-2.5g/L, magnesium sulfate 0.1-0.75g/L, L-
Cysteine hydrochloride 0.001-0.1g/L, Tween-20 0.5-2.0mL/L, pH 6.0-7.0.
Antioxidant in the step (b) is sodium isoascorbate, additive amount 8-15g/L.
Sterilising conditions in the step (b) are 115 DEG C × 15min.
Inoculum density in the step (c) is calculated as 0.2g/L with bifidobacterium bifidum bacterium powder.
Antioxidant in the step (d) is sodium ascorbate.
The additive amount of antioxidant is the 0.8-1.5 of antioxidant additive amount quality in step (a) in the step (d)
Times.
In the step (d), when viable count is 3-4.5 × 107Antioxidant is added when cfu/mL.
The accumulative culture 48h of the step (e) terminates to cultivate.
The present invention provides a kind of industrial processes of beneficial bacteria of intestinal tract preparation, comprising the following steps: (1) will do respectively
Lactobacillus paracasei, Lactobacillus rhamnosus, bifidobacterium bifidum, lactobacillus delbruockii subspecies bulgaricus are inoculated into different fermented and cultureds
Fermented and cultured is carried out in base, obtains fermentation culture；(2) streptococcus thermophilus is inoculated into fermentation medium and carries out fermentation training
It supports, adds porous-starch before fermentation ends into fermentation liquid, obtain fermentation culture；(3) collection step (1) and step (2) institute
The fermentation liquid of each strain obtained obtains the wet bacterium mud of activity, adds distilled water to be uniformly mixed with protective agent wet bacterium mud；(4) by granulation
Machine, spheronizator, drying machine, seed-coating machine pelletized, be round as a ball, low-temperature vacuum drying, coating, obtains enteron aisle after detection is qualified
Probiotics preparation.It is able to ascend the quantity of viable bacteria in probiotics preparation using method of the invention, extends depositing for probiotics preparation
It puts the time, reduces production cost.
The present invention provides a kind of industrial process of beneficial bacteria of intestinal tract preparation, this method is in lower protective agent additive amount
On the basis of, it can guarantee higher Viable detection, significantly improve the content of viable bacteria in every gram of finished product.
The present invention provides a kind of industrial processes of beneficial bacteria of intestinal tract preparation, the described method comprises the following steps:
(1) respectively by Lactobacillus casei, Lactobacillus rhamnosus, bifidobacterium bifidum, lactobacillus delbruockii subspecies bulgaricus
It is inoculated into different fermentation mediums and carries out fermented and cultured, when viable count is up to standard in fermented and cultured, terminates fermented and cultured, obtain
To fermentation culture；
(2) streptococcus thermophilus is inoculated into fermentation medium and carries out fermented and cultured, added before fermentation ends into fermentation liquid
Add porous-starch, obtains fermentation culture；
(3) fermentation liquid of collection step (1) and the resulting each strain of step (2) and mixing, are centrifuged and are lived after washing
Property wet bacterium mud, add distilled water to be uniformly mixed with protective agent wet bacterium mud, obtain bacterium mud protection agent composition, wherein wet bacterium mud: protecting
Protect agent: the mass ratio of distilled water is 1:0.5-0.8:0.4-0.8；
(4) the obtained bacterium mud protection agent composition of step (3) is successively passed through into granulator, spheronizator, drying machine, coating
Machine pelletized, is round as a ball, low-temperature vacuum drying, coating, obtains beneficial bacteria of intestinal tract preparation after detection is qualified.
Fermentative medium formula in the step (1) are as follows: glucose 5.0-15.0g, pancreatin hydrolysis casein 2.0-
15.0g, yeast extract 1.5-5.5g, sodium citrate 1.0-5.0g, dipotassium hydrogen phosphate 0.2-2.5g, magnesium sulfate 0.1-0.75g, L- half
Cystine hydrochloride 0.001-0.1g, Tomato juice 2.0-25g, Tween-20 0.5-2.0mL, Jia Shui are settled to 1000mL, pH
6.0-7.0,105 DEG C sterilize 15 minutes.
The step (1) when in fermentation liquid viable count reach 3-4.5 × 109When cfu/mL, terminate fermented and cultured.
Fermentation medium in the step (2) are as follows: glucose 95-98g/L, ammonium sulfate 7-10g/L, peptone 95-99g/
L, ammonium nitrate 4-8g/L, potassium dihydrogen phosphate 3-4g/L, magnesium sulfate 1-2g/L, pH 6.8-7.1,105 DEG C sterilize 15 minutes.
The step (2) when in fermentation liquid viable count reach 2-3 × 109When cfu/mL, porous-starch is added, continues to ferment
Culture 1-5 hours.
The step (2) adds porous-starch, makes it in initial content 10-20g/L.
Cultivation temperature in step (1) and step (2) is 30 DEG C -45 DEG C.
Protective agent in the step (3) includes each substance of following mass parts: 1-2 parts of glycerol, 2-3 parts of oligomeric different malt
Sugar, 2-4.5 parts of oligofructose, 0.1-2.5 parts of soybean peptides, 2-8 parts of calcium silicates.
In some preferred technical solutions, the brevibacterium flavum in the protective agent also containing the inactivation of 1-2 mass parts is sent out
Zymotic fluid.
The preparation method of the brevibacterium flavum fermentation liquid of the inactivation is that culture medium includes: glucose 20-40g/L, yeast
Powder 2-8g/L, ammonium sulfate 2-8g/L, Dried Corn Steep Liquor Powder 15-25g/L, potassium dihydrogen phosphate 0.5-3g/L, magnesium sulfate 0.8-1.5g/L；
PH is 6.9-7.2；Inoculum concentration is calculated as 0.1g/L with brevibacterium flavum bacterium powder；30 DEG C -45 DEG C of cultures 3-8 hours；High-temperature sterilization.
Compared with prior art, the invention has the benefit that
(1) present invention is reduced by before fermentation ends, adding porous-starch into the fermentation medium of streptococcus thermophilus
The addition of porous-starch in wet bacterium mud, and then increase the quantity of viable bacteria in every gram of product.
(2) fermentation liquid of the invention by adding a certain amount of brevibacterium flavum through inactivating in beneficial bacteria of intestinal tract, is improved
The survival rate of probiotics in final product.
(3) present invention is by adding porous-starch in the streptococcus thermophilus fermentation later period simultaneously and adding yellow in wet bacterium mud
Quarter butt fermented liquid extends the resting period of beneficial bacteria of intestinal tract preparation.
(4) present invention is it has unexpectedly been discovered that it is prebiotic that calcium silicates can increase every gram of enteron aisle on the basis of inventive formulation
Number of viable in bacteria preparation.
Specific embodiment
Unless otherwise defined, all technical and scientific terms used herein have and the technical field of the invention
The normally understood identical meaning of those of ordinary skill.In the examples where no specific technique or condition is specified, according in the art
Conventional technical means carries out.
A kind of chemical stress removal method of High Density Cultivation bifidobacterium bifidum of embodiment a
A. the culture medium of bifidobacterium bifidum, glucose 5.0g/L, pancreatin hydrolysis casein 2.0g/L, corn pulp are prepared
1.5g/L, sodium citrate 1.0g/L, dipotassium hydrogen phosphate 0.2g/L, magnesium sulfate 0.1g/L, L-cysteine hydrochloride 0.001g/L,
Tween-20 0.5mL/L, pH 6；
B. addition oxidation preventive isoascorbic acid sodium in the culture medium that step (a) is prepared, additive amount 8g/L, and 115 DEG C
× 15min carries out sterilization treatment；
C. it is inoculated with, bifidobacterium bifidum bacterium powder is calculated as 0.2g/L；
D. when viable count is 3 × 10 in incubation7Continue to add antioxidants ascorbic acid sodium, additive amount when cfu/mL
It is 0.8 times of sodium isoascorbate addition quality；
E. after cultivating 48h, count plate is carried out using colony counting method.
A kind of chemical stress removal method of High Density Cultivation bifidobacterium bifidum of embodiment b
A. the culture medium of bifidobacterium bifidum, glucose 15.0g/L, pancreatin hydrolysis casein 15.0g/L, corn pulp are prepared
5.5g/L, sodium citrate 5.0g/L, dipotassium hydrogen phosphate 2.5g/L, magnesium sulfate 0.75g/L, L-cysteine hydrochloride 0.1g/L,
Tween-20 2.0mL/L, pH7；
B. addition oxidation preventive isoascorbic acid sodium in the culture medium that step (a) is prepared, additive amount 15g/L, and 115 DEG C
× 15min carries out sterilization treatment；
C. it is inoculated with, bifidobacterium bifidum bacterium powder is calculated as 0.2g/L；
D. when viable count is 4.5 × 10 in incubation7Continue to add antioxidants ascorbic acid sodium when cfu/mL, add
Amount is 1.5 times of sodium isoascorbate addition quality；
E. after cultivating 48h, count plate is carried out using colony counting method.
A kind of industrial process of the beneficial bacteria of intestinal tract preparation of embodiment 1
(1) respectively by Lactobacillus casei, Lactobacillus rhamnosus, bifidobacterium bifidum, lactobacillus delbruockii subspecies bulgaricus
It is inoculated into different fermentation mediums and carries out fermented and cultured, when viable count reaches 4.5 × 10 in fermented and cultured9When cfu/mL,
Terminate fermented and cultured, obtain fermentation culture, wherein cultivation temperature is 30 DEG C DEG C, and the formula of fermentation medium is equal are as follows: glucose
15.0g, pancreatin hydrolysis casein 2.0g, yeast extract 5.5g, sodium citrate 1.0g, dipotassium hydrogen phosphate 2.5g, magnesium sulfate 0.1g, L-
Cysteine hydrochloride 0.1g, Tomato juice 2.0g, Tween-20 2.0mL, Jia Shui are settled to 6.0,105 DEG C of 1000mL, pH and go out
Bacterium 15 minutes；
(2) streptococcus thermophilus is inoculated into fermentation medium and carries out fermented and cultured, when in fermentation liquid viable count reach 3 ×
109When cfu/mL, porous-starch is added into fermentation liquid, makes initial content 10g/L of the porous-starch in fermented and cultured, after
Continuous fermented and cultured 5 hours, fermentation culture is obtained, cultivation temperature is 30 DEG C, fermentation medium used are as follows: glucose 98g/L, sulphur
Sour ammonium 7g/L, peptone 99g/L, ammonium nitrate 4g/L, potassium dihydrogen phosphate 4g/L, 7.1,105 DEG C of magnesium sulfate 1g/L, pH sterilizings 15
Minute；
(3) fermentation liquid of collection step (1) and the resulting each strain of step (2) and mixing, are centrifuged and are lived after washing
Property wet bacterium mud, add distilled water to be uniformly mixed with protective agent wet bacterium mud, obtain bacterium mud protection agent composition, wherein wet bacterium mud: protecting
Protect agent: the mass ratio of distilled water be 1:0.5:0.8, the protective agent include following mass parts each substance: 1 part of glycerol, 3 parts it is low
Polyisomaltose, 2 parts of oligofructose, 2.5 parts of soybean peptides, 2 parts of calcium silicates；
(4) the obtained bacterium mud protection agent composition of step (3) is successively passed through into granulator, spheronizator, drying machine, coating
Machine pelletized, is round as a ball, low-temperature vacuum drying, coating, obtains beneficial bacteria of intestinal tract preparation after detection is qualified.
A kind of industrial process of the beneficial bacteria of intestinal tract preparation of embodiment 2
(1) respectively by Lactobacillus casei, Lactobacillus rhamnosus, bifidobacterium bifidum, lactobacillus delbruockii subspecies bulgaricus
It is inoculated into different fermentation mediums and carries out fermented and cultured, when viable count reaches 3 × 10 in fermented and cultured9When cfu/mL, knot
Beam fermented and cultured, obtains fermentation culture, wherein cultivation temperature is 45 DEG C, and the formula of fermentation medium is equal are as follows: glucose
5.0g, pancreatin hydrolysis casein 15.0g, yeast extract 1.5g, sodium citrate 5.0g, dipotassium hydrogen phosphate 0.2g, magnesium sulfate 0.75g,
L-cysteine hydrochloride 0.001g, Tomato juice 25g, Tween-20 0.5mL, Jia Shui are settled to 1000mL, pH 7.0,105
DEG C sterilizing 15 minutes；
(2) streptococcus thermophilus is inoculated into fermentation medium and carries out fermented and cultured, when in fermentation liquid viable count reach 2 ×
109When cfu/mL, porous-starch is added into fermentation liquid, makes initial content 20g/L of the porous-starch in fermented and cultured, after
Continuous fermented and cultured 1 hour, fermentation culture is obtained, cultivation temperature is 45 DEG C, fermentation medium used are as follows: glucose 95g/L, sulphur
Sour ammonium 10g/L, peptone 95g/L, ammonium nitrate 8g/L, potassium dihydrogen phosphate 3g/L, 6.8,105 DEG C of magnesium sulfate 2g/L, pH sterilizings 15
Minute；
(3) fermentation liquid of collection step (1) and the resulting each strain of step (2) and mixing, are centrifuged and are lived after washing
Property wet bacterium mud, add distilled water to be uniformly mixed with protective agent wet bacterium mud, obtain bacterium mud protection agent composition, wherein wet bacterium mud: protecting
Protect agent: the mass ratio of distilled water be 1:0.8:0.4, the protective agent include following mass parts each substance: 2 parts of glycerol, 2 parts it is low
Polyisomaltose, 4.5 parts of oligofructose, 0.1 part of soybean peptide, 8 parts of calcium silicates；
(4) the obtained bacterium mud protection agent composition of step (3) is successively passed through into granulator, spheronizator, drying machine, coating
Machine pelletized, is round as a ball, low-temperature vacuum drying, coating, obtains beneficial bacteria of intestinal tract preparation after detection is qualified.
A kind of industrial process of the beneficial bacteria of intestinal tract preparation of embodiment 3
(1) respectively by Lactobacillus casei, Lactobacillus rhamnosus, bifidobacterium bifidum, lactobacillus delbruockii subspecies bulgaricus
It is inoculated into different fermentation mediums and carries out fermented and cultured, when viable count reaches 3.5 × 10 in fermented and cultured9When cfu/mL,
Terminate fermented and cultured, obtain fermentation culture, wherein cultivation temperature is 40 DEG C, and the formula of fermentation medium is equal are as follows: glucose
10.0g, pancreatin hydrolysis casein 8.0g, yeast extract 4.2g, sodium citrate 4.0g, dipotassium hydrogen phosphate 2.1g, magnesium sulfate 0.5g, L-
Cysteine hydrochloride 0.006g, Tomato juice 15g, Tween-20 1.0mL, Jia Shui are settled to 6.5,105 DEG C of 1000mL, pH
Sterilizing 15 minutes；
(2) streptococcus thermophilus is inoculated into fermentation medium and carries out fermented and cultured, when viable count reaches 2.3 in fermentation liquid
×109When cfu/mL, porous-starch is added into fermentation liquid, makes initial content 15g/L of the porous-starch in fermented and cultured,
Continue fermented and cultured 3 hours, obtain fermentation culture, cultivation temperature is 40 DEG C, fermentation medium used are as follows: glucose 97g/L,
Ammonium sulfate 8g/L, peptone 97g/L, ammonium nitrate 7g/L, potassium dihydrogen phosphate 3.5g/L, 7.0,105 DEG C of magnesium sulfate 1.2g/L, pH
Sterilizing 15 minutes；
(3) fermentation liquid of collection step (1) and the resulting each strain of step (2) and mixing, are centrifuged and are lived after washing
Property wet bacterium mud, add distilled water to be uniformly mixed with protective agent wet bacterium mud, obtain bacterium mud protection agent composition, wherein wet bacterium mud: protecting
Protect agent: the mass ratio of distilled water is 1:0.6:0.7, and the protective agent includes each substance of following mass parts: 1.5 parts of glycerol, 2.1
Part oligoisomaltose, 3.0 parts of oligofructose, 1.2 parts of soybean peptides, 6 parts of calcium silicates；
(4) the obtained bacterium mud protection agent composition of step (3) is successively passed through into granulator, spheronizator, drying machine, coating
Machine pelletized, is round as a ball, low-temperature vacuum drying, coating, obtains beneficial bacteria of intestinal tract preparation after detection is qualified.
A kind of industrial process of the beneficial bacteria of intestinal tract preparation of embodiment 4
Difference with embodiment 1 is only that protective agent used in step (3) includes each substance of following mass parts: 1 part of glycerol,
3 parts of oligoisomaltoses, 2 parts of oligofructose, 2.5 parts of soybean peptides, 2 parts of calcium silicates, 1 portion of brevibacterium flavum fermentation liquid inactivated,
The preparation method of the brevibacterium flavum fermentation liquid of the inactivation is that culture medium includes: glucose 40g/L, yeast powder
2g/L, ammonium sulfate 8g/L, Dried Corn Steep Liquor Powder 15g/L, potassium dihydrogen phosphate 3g/L, magnesium sulfate 0.8g/L, pH 7.2, inoculum concentration with
Brevibacterium flavum bacterium powder is calculated as 0.1g/L, and 30 DEG C are cultivated 8 hours, high-temperature sterilization.
A kind of industrial process of the beneficial bacteria of intestinal tract preparation of embodiment 5
Difference with embodiment 2 is only that protective agent used in step (3) includes each substance of following mass parts: 2 parts of glycerol,
2 parts of oligoisomaltoses, 4.5 parts of oligofructose, 0.1 part of soybean peptide, 8 parts of calcium silicates, 2 parts of brevibacterium flavum fermentations inactivated
Liquid,
The preparation method of the brevibacterium flavum fermentation liquid of the inactivation is that culture medium includes: glucose 20g/L, yeast powder
8g/L, ammonium sulfate 2g/L, Dried Corn Steep Liquor Powder 25g/L, potassium dihydrogen phosphate 0.5g/L, magnesium sulfate 1.5g/L, pH 6.9, inoculum concentration
It is calculated as 0.1g/L with brevibacterium flavum bacterium powder, 45 DEG C are cultivated 3 hours, high-temperature sterilization.
A kind of industrial process of the beneficial bacteria of intestinal tract preparation of embodiment 6
Difference with embodiment 3 is only that protective agent used in step (3) includes each substance of following mass parts: 1.5 parts sweet
Oil, 2.1 parts of oligoisomaltoses, 3.0 parts of oligofructose, 1.2 parts of soybean peptides, 6 parts of calcium silicates, 1.5 parts of yellow quarter butts inactivated
Fermented liquid；
The preparation method of the brevibacterium flavum fermentation liquid of the inactivation is that culture medium includes: glucose 30g/L, yeast powder
6g/L, ammonium sulfate 4g/L, Dried Corn Steep Liquor Powder 20g/L, potassium dihydrogen phosphate 1.5g/L, magnesium sulfate 1.1g/L；PH is 7.0；Inoculum concentration
0.1g/L is calculated as with brevibacterium flavum bacterium powder；40 DEG C are cultivated 5 hours；High-temperature sterilization.
A kind of industrial process of the beneficial bacteria of intestinal tract preparation of comparative example 1
Difference with embodiment 3 is only that the porous-starch added in step (2), which is put into step (3), neutralizes protective agent one
Addition is played, wherein the amount of the porous-starch added and protectant amount are equal with the corresponding additive amount in embodiment 3 respectively.
A kind of industrial process of the beneficial bacteria of intestinal tract preparation of comparative example 2
Difference with embodiment 6 is only that the porous-starch added in step (2), which is put into step (3), neutralizes protective agent one
Addition is played, wherein the amount of the porous-starch added and protectant amount are equal with the corresponding additive amount in embodiment 6 respectively.
A kind of industrial process of the beneficial bacteria of intestinal tract preparation of comparative example 3
Chinese patent announces preparation method shown in embodiment 3 in CN108783462A.
Viable count statistics: beneficial bacteria of intestinal tract preparation 0.1g made from Example 1-6 and comparative example 1-3 is dissolved in appropriate nothing
(dilution 10 in bacterium water7), count plate is carried out after dilution, as a result as shown in table 1 below.
Resting period statistics: by beneficial bacteria of intestinal tract preparation made from embodiment 1-6 and comparative example 1-3, temperature 37 is deposited in
It ± 2 DEG C, in the environment of humidity 75 ± 5%, in 0th month, first month and second month, unites to the viable count in preparation
Meter, and calculates the survival rate of probiotics, and the viable count at viable count/0th the end of month at survival rate=the first or two the end of month ×
100%, it the results are shown in Table 1.
Table 1
Experimental data in table 1 is shown, prepares beneficial bacteria of intestinal tract preparation using method of the invention, is able to ascend every gram of production
The quantity of viable bacteria in product, reduces protectant addition, improves the economic benefit of production.By embodiment 1-3 successively with embodiment 4-6
It is compared, it is found that on the basis of inventive method, the brevibacterium flavum fermentation liquid of inactivation, energy are added in protective agent
Enough number of viable further promoted in every gram of beneficial bacteria of intestinal tract preparation, and the addition energy of the brevibacterium flavum fermentation liquid inactivated
Inactivation rate of probiotics during preservation is enough reduced, the holding time is extended.
Claims (10)
1. a kind of chemical stress removal method of High Density Cultivation bifidobacterium bifidum, which comprises the following steps:
A. the culture medium of bifidobacterium bifidum is prepared；
B. antioxidant is added in the culture medium that step (a) is prepared, and carries out sterilization treatment；
C. it is inoculated with；
D. continue to add antioxidant in incubation；
E. after cultivating, count plate is carried out using colony counting method.
2. chemical stress removal method according to claim 1, which is characterized in that the culture medium in the step (a) are as follows: grape
Sugared 5.0-15.0g/L, pancreatin hydrolysis casein 2.0-15.0g/L, corn pulp 1.5-5.5g/L, sodium citrate 1.0-5.0g/L,
Dipotassium hydrogen phosphate 0.2-2.5g/L, magnesium sulfate 0.1-0.75g/L, L-cysteine hydrochloride 0.001-0.1g/L, Tween-20
0.5-2.0mL/L, pH 6.0-7.0.
3. chemical stress removal method according to claim 1, it is characterised in that: the antioxidant in the step (b) is different
Sodium ascorbate, additive amount 8-15g/L.
4. chemical stress removal method according to claim 1, it is characterised in that: the sterilising conditions in the step (b) are 115
℃×15min。
5. chemical stress removal method according to claim 1, it is characterised in that: the inoculum density in the step (c), with two
Discrimination Bifidobacteria powder is calculated as 0.2g/L.
6. chemical stress removal method according to claim 1, it is characterised in that: the antioxidant in the step (d) is anti-
Bad hematic acid sodium.
7. chemical stress removal method according to claim 1, it is characterised in that: the addition of antioxidant in the step (d)
Amount is 0.8-1.5 times of antioxidant additive amount quality in step (a).
8. chemical stress removal method according to claim 1, it is characterised in that: in the step (d), when viable count is 3-
4.5×107Antioxidant is added when cfu/mL.
9. chemical stress removal method according to claim 1, it is characterised in that: the accumulative culture 48h of the step (e) terminates to train
It supports.
10. the bifidobacterium bifidum of the described in any item chemical stress removal method cultures of claim 1-9 is in beneficial bacteria of intestinal tract preparation
In application.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111534566A (en) * | 2020-06-17 | 2020-08-14 | 江南大学 | A gradient dilution and its application in counting bifidobacteria |
| CN112553116A (en) * | 2020-12-23 | 2021-03-26 | 邢箫 | Probiotic culture medium and probiotic preparation |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103333840A (en) * | 2013-07-08 | 2013-10-02 | 昆山佰生优生物科技有限公司 | Probiotic ultralow temperature refrigeration technology and applications thereof in probiotic preparation |
| CN108323761A (en) * | 2017-12-27 | 2018-07-27 | 深圳市麦金利实业有限公司 | A kind of composite reactive probiotics preparation and preparation method thereof |
| CN108783462A (en) * | 2018-07-02 | 2018-11-13 | 哈尔滨美华生物技术股份有限公司 | A kind of industrial process of beneficial bacteria of intestinal tract preparation |
| CN108902315A (en) * | 2018-07-12 | 2018-11-30 | 广州华农大食品科技有限公司 | A kind of probiotic mixed fermentation Yoghourt and production method |
-
2019
- 2019-02-14 CN CN201910114816.XA patent/CN109593690B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103333840A (en) * | 2013-07-08 | 2013-10-02 | 昆山佰生优生物科技有限公司 | Probiotic ultralow temperature refrigeration technology and applications thereof in probiotic preparation |
| CN108323761A (en) * | 2017-12-27 | 2018-07-27 | 深圳市麦金利实业有限公司 | A kind of composite reactive probiotics preparation and preparation method thereof |
| CN108783462A (en) * | 2018-07-02 | 2018-11-13 | 哈尔滨美华生物技术股份有限公司 | A kind of industrial process of beneficial bacteria of intestinal tract preparation |
| CN108902315A (en) * | 2018-07-12 | 2018-11-30 | 广州华农大食品科技有限公司 | A kind of probiotic mixed fermentation Yoghourt and production method |
Non-Patent Citations (1)
| Title |
|---|
| 宫卫波: "《黄色短杆菌ilvBN 和ilvC 基因定点突变对L-缬氨酸发酵的影响》", 《发酵科技通讯》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111534566A (en) * | 2020-06-17 | 2020-08-14 | 江南大学 | A gradient dilution and its application in counting bifidobacteria |
| CN112553116A (en) * | 2020-12-23 | 2021-03-26 | 邢箫 | Probiotic culture medium and probiotic preparation |
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