CN108841753A - A kind of lactic acid bacteria high density fermentation culture medium, preparation method and applications - Google Patents
A kind of lactic acid bacteria high density fermentation culture medium, preparation method and applications Download PDFInfo
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Abstract
The present invention provides a kind of lactic acid bacteria high density fermentation culture mediums, preparation method and applications, wherein, culture medium is made of protein enzymatic hydrolyzate, yeast powder and partial salts, and enzymolysis liquid is made of Alcalase protease hydrolytic skimmed milk powder and soybean protein isolate, and specific formula is：Skimmed milk powder (6%), soybean protein isolate (2%), glucose (3%), pearling cone meal (2%), yeast powder (1.8%), NaCl (0.23%), VC (0.3%), MgSO4·7H2O (0.02%), MnSO4·H2O (0.005%), sodium citrate (0.02%).Lactobacillus plantarum, Lactobacillus rhamnosus and lactobacillus paracasei High Density Cultivation are carried out using basal culture medium, until culture stationary phase culture solution viable count can reach 1~1.5 × 1010Cfu/mL can effectively improve the viable count of lactic acid bacteria High Density Cultivation.
Description
Technical field
The present invention relates to field of microbial fermentation, and in particular to a kind of lactic acid bacteria high density fermentation culture medium, preparation method
And its application.
Background technique
Lactic acid bacteria is the logical of a kind of gram-positive bacterium that a large amount of lactic acid can be generated using fermentable carbohydrate
Claim.It is widely present in people, animal, fowl enteron aisle, numerous food product, material and a small number of clinical samples.As domestic and foreign scholars are to lactic acid
The further investigation of bacterium finds it with reducing blood lipid, height-regulating immunity, adjusting intestinal microecology balance, antitumor, treatment diabetes
Etc. physiological functions.Meanwhile with the continuous development of biotechnology, lactic acid bacteria shows in the light industrys industry such as food, medicine, animal husbandry
Good application prospect out.
With high-cell-density cultivation (High Cell Density Culture, HCDC), separation and Freeze Drying Technique
Development, standardization, high stability probiotics freeze-dried vaccine powder provide bridge and guarantee for the extensive use of probiotics.Therefore
Also become probiotics work for how to improve probiotic's culture liquid viable count, freeze-drying survival rate, production efficiency and storage stability
The hot spot of industry PRODUCTION TRAITS.And whether the primary condition that culture medium is grown as microorganism, composition will properly directly affect training
The proliferation and activity of lactic acid bacteria during supporting.
And the streptococcus acidi lactici fermented solution of high concentration is obtained by High Density Cultivation technology, it is necessary to optimize a series of relevant
Technological parameter carries out stringent control to fermentation process.By solving these problems, designs suitable ways for training and reach
Ideal cell concentration.The cause that research report shows to influence lactic acid bacteria proliferation be known as it is very much, as strain vigor, cultivate algebra,
Cultivation cycle, the initial pH value of culture solution, cultivation temperature, incubation time, inoculum concentration, proliferation factor etc., the wherein object of culture medium
Rationality matter and the collection work that lactic acid bacteria thallus is seriously affected at branch, so the selection multiselect viscosity of culture medium is small, egg
The low transparent liquid of white matter content, cultivation temperature and incubation time can all influence the vigor of strain.And the proliferation of lactic acid bacteria because
Son is also relatively more, as tomato juice, carrot juice, mushroom juice, soybean enzymolysis liquid etc. can all promote the numerous of thallus to a certain extent
Reproductive growth.Therefore, it is just heavy to closing for the streptococcus acidi lactici fermented solution for obtaining high concentration, high activity to develop a kind of suitable culture medium
Want, at the same the lactobacillus-fermented of early period also by the later period freeze-drying and storage have an important influence on.
Summary of the invention
In view of this, the present invention provides a kind of lactic acid bacteria high density fermentation culture medium, preparation method and applications.
First aspect present invention provides a kind of preparation method of lactic acid bacteria high density fermentation culture medium, including following step
Suddenly：
(1) protein enzymatic hydrolyzate raw material, pearling cone meal enzymolysis liquid raw material and the basal medium of following weight percent are provided
Raw material, wherein the raw material of the protein enzymatic hydrolyzate includes skimmed milk powder (2~10%), soybean protein isolate (1~3%)；Barley
The raw material of powder enzymolysis liquid includes pearling cone meal (1~3%)；
(2) prepared by protein enzymatic hydrolyzate：Skimmed milk powder, the soybean protein isolate of the offer of step (1) are dissolved in water, enzyme enzyme
Solve protein enzymatic hydrolyzate；
(3) prepared by basal medium：The basal medium raw material of the offer of step (1) is dissolved in water, obtains basal medium；
(4) prepared by pearling cone meal enzymolysis liquid：The pearling cone meal of the offer of step (1) is dissolved in water, it is enzyme to digest to obtain pearling cone meal
Enzymolysis liquid；
(5) protein enzymatic hydrolyzate, basal medium and pearling cone meal enzymolysis liquid are mixed, constant volume, high-temperature sterilization, is cooled down cold
But, pH value is adjusted, lactic acid bacteria high density fermentation culture medium is obtained.
In an embodiment of the invention, in the preparation method step (1), basal medium raw material includes：Grape
Sugared (1~5%), yeast powder (1~3%), NaCl (0.1~0.3%), VC (0.2~0.4%), MgSO47H2O (0.01~
0.03%), MnSO4H2O (0.003~0.007%), sodium citrate (0.01~0.03%).
In an embodiment of the invention, the enzyme being added in the preparation method step (2) is protease, additional amount
It is the 0.1~1% of protein enzymatic hydrolyzate raw material；
In a specific embodiment of the invention, in the preparation method step (2), the enzyme of addition is Alcalase egg
White enzyme, additional amount 0.2%.
In an embodiment of the invention, in the preparation method step (2), the condition of enzymatic hydrolysis is temperature 30~60
DEG C, it is kept for 1~8 hour under 60~100rpm；
In a specific embodiment of the invention, in the preparation method step (2), the condition of enzymatic hydrolysis is temperature 45
DEG C, it is kept for 4 hours under 80rpm.
In an embodiment of the invention, in the preparation method step (4), the enzyme of addition is barley stickiness polysaccharide water
Enzyme and beta amylase are solved, additional amount is the 0.1~1% of pearling cone meal enzymolysis liquid raw material；
In a specific embodiment of the invention, in the preparation method step (4), the enzyme of addition is that barley stickiness is more
Glycosylhydrolase and beta amylase, additional amount are the 0.1% of pearling cone meal enzymolysis liquid raw material.
In an embodiment of the invention, in the preparation method step (4), the pH value of culture medium is 6.0~7.0；
In a specific embodiment of the invention, in the preparation method step (5), the pH value of culture medium is 6.0.
Third aspect present invention provides a kind of lactic acid bacteria high density fermentation culture medium, such as provided such as first aspect present invention
Application of the preparation method for the lactic acid bacteria high density fermentation culture medium that second aspect of the present invention provides in lactobacillus-fermented.
In an embodiment of the invention, the application includes the following steps：The cream that first aspect present invention is provided
Sour bacterium high density fermentation culture medium is proportionally added into the lactobacillus solution activated, adjusts pH value, then ferments, obtain high density
The streptococcus acidi lactici fermented solution of fermentation.
In a specific embodiment of the invention, the additional proportion of the lactic acid bacteria high density fermentation culture medium is lactic acid
The 5~20% of bacterium seed liquor；
Preferably, the additional proportion of the lactic acid bacteria high density fermentation culture medium is the 10% of lactobacillus solution.
In a specific embodiment of the invention, the lactobacillus solution is after being added high density fermentation culture medium,
It is 6.0 that pH value, which need to be adjusted,.
In a specific embodiment of the invention, the fermentation temperature of the high density fermentation is 30~40 DEG C, and fermentation is extremely
Fermentation liquid OD600In stable state, the streptococcus acidi lactici fermented solution of high density fermentation is obtained.
High Density Cultivation technology is widely used in the production of various microorganisms, it refers to using certain technique skill
Art, guarantee microorganism growth suitable condition, extend the exponential growth process of microorganism, thus obtain high concentration cell and
Metabolite.The High Density Cultivation of lactic acid bacteria increases the thalline quantity in unit volume culture solution, reduces production cost, from
And improve product competitiveness in the market.
The present invention is added to skimmed milk powder, soybean protein isolate inside the component of lactic acid bacteria high density fermentation culture medium,
And skimmed milk powder, soybean protein isolate are digested at the small molecule more suitable for lactic acid bacteria high density fermentation, together using protease
When be added to pearling cone meal, and pearling cone meal is resolved into more suitable for lactic acid bacteria height using barley stickiness polysaccharide hydrolase beta amylase
The small molecule of density fermentation, to obtain to effectively improve the lactic acid bacteria high density hair of lactic acid bacterial liquid concentration and lactic bacteria activity
Ferment culture medium.
The raw material sources of culture medium provided by the invention are extensive, it is low in cost, be easy to transport and store, and prepare it is simple,
It can scale use；Meanwhile using the culture medium, lactic acid bacteria high density fermentation bacterial concentration can be effectively improved and lactic acid bacteria is living
Property, fermentation costs are reduced, production efficiency is improved, are had a good application prospect.
Specific embodiment
The following is a preferred embodiment of the present invention, it is noted that for those skilled in the art
For, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also considered as
Protection scope of the present invention.
Below by a preferred embodiment of the present invention will be described in detail.The experiment of actual conditions is not specified in embodiment
Method, usually according to normal condition.In the embodiment of the present invention unless otherwise noted, agents useful for same and consumptive material are commercial goods.
The present invention provides a kind of lactic acid bacteria high density fermentation culture medium, and component includes：Skimmed milk powder (6%), soybean point
From albumen (2%), glucose (3%), pearling cone meal (2%), yeast powder (1.8%), NaCl (0.23%), VC (0.3%),
MgSO4·7H2O (0.02%), MnSO4·H2O (0.005%), sodium citrate (0.02%).
Lactic acid bacteria high density fermentation culture medium preparation method provided by the invention, includes the following steps：
(1) weigh skimmed milk powder respectively in proportion, soybean protein isolate is dissolved in suitable water, 0.2% is added at this time
Alcalase protease, control temperature 4h is kept under 45 DEG C, 80rpm, acquired solution is protein enzymatic hydrolyzate；
(2) glucose, yeast powder, NaCl, VC, MgSO are weighed in proportion4·7H2O、MnSO4·H2O, sodium citrate is abundant
It is dissolved in suitable quantity of water, as basic culture medium；
(3) prepared by pearling cone meal enzymolysis liquid：Pearling cone meal is weighed in proportion to be dissolved in suitable water, and 0.1% is added at this time
Barley stickiness polysaccharide hydrolase and 0.1% beta amylase, control temperature 4h is kept under 55 DEG C, 80rpm, acquired solution is
Pearling cone meal enzymolysis liquid；
(4) protein enzymatic hydrolyzate, basal medium and pearling cone meal enzymolysis liquid are mixed, then constant volume, at 115 DEG C,
Sterilize 20min, and then cooling down adjusts pH to 6.0 with sterilized NaOH to 37 DEG C, can be obtained lactic acid bacteria high density hair
Ferment culture medium.
A kind of lactic acid bacteria high density fermentation culture medium provided by the invention can be used for lactobacillus plantarum, Lactobacillus rhamnosus,
The High Density Cultivation of the lactic acid bacterias such as lactobacillus paracasei, specific embodiment are as follows：
Embodiment 1：Lactobacillus plantarum High Density Cultivation
(1) preparation of culture medium
Culture medium A：It is skimmed milk powder (6%), soybean protein isolate (2%), glucose (3%), yeast powder (1.8%), big
Flour (2%), NaCl (0.23%), VC (0.3%), MgSO4·7H2O (0.02%), MnSO4·H2O (0.005%), lemon
Sour sodium (0.02%).
Preparation method：Skimmed milk powder, soybean protein isolate are added in proportion first and had warmed up into 45 DEG C of water, benefit
It is stirred with blender to abundant dissolution, then protein liquid is putted into fermenter, 0.1% Alcalase protease is added, controlled
Temperature processed keeps 4h under 45 DEG C, 80rpm, weighs glucose, yeast powder, NaCl, VC, MgSO in proportion later4·7H2O、
MnSO4·H2O, sodium citrate is completely dissolved in suitable quantity of water and puts into fermenter, and finally digests the pearling cone meal digested in advance
Liquid is added in fermentor, adds water constant volume to mix, sterilize 20min at 115 DEG C, after being cooled to 37 DEG C, with sterilized NaOH tune
Save pH to 6.0.
Culture medium B：Skimmed milk powder (6%), soybean protein isolate (2%), glucose (3%), yeast powder (1.8%),
NaCl (0.23%), VC (0.3%), MgSO4·7H2O (0.02%), MnSO4·H2O (0.005%), sodium citrate
(0.02%).
Preparation method：Weigh in proportion skimmed milk powder, soybean protein isolate, glucose, yeast powder, NaCl, VC,
MgSO4·7H2O、MnSO4·H2O, sodium citrate is completely dissolved in suitable quantity of water and puts into fermenter, and water constant volume is added to mix, in
Sterilize 20min at 115 DEG C, after being cooled to 37 DEG C, adjusts pH to 6.0 with sterilized NaOH.
Culture medium C：Skimmed milk powder (6%), soybean protein isolate (2%), glucose (3%), yeast powder (1.8%),
NaCl (0.23%), VC (0.3%), MgSO4·7H2O (0.02%), MnSO4·H2O (0.005%), sodium citrate
(0.02%).
Preparation method：Skimmed milk powder and soybean protein isolate are added in proportion first and had warmed up into 45 DEG C of water,
It is stirred using blender to abundant dissolution, then protein liquid is putted into fermenter, 0.1% Alcalase protease is added,
Control temperature keeps 4h under 45 DEG C, 80rpm, weighs glucose, yeast powder, NaCl, VC, MgSO in proportion later4·7H2O、
MnSO4·H2O, sodium citrate is completely dissolved in suitable quantity of water and puts into fermenter, and adds water constant volume to mix, sterilizes at 115 DEG C
20min after being cooled to 37 DEG C, adjusts pH to 6.0 with sterilized NaOH.
Culture medium D：Skimmed milk powder (8%), glucose (3%), yeast powder (1.8%), NaCl (0.23%), VC
(0.3%), MgSO4·7H2O (0.02%), MnSO4·H2O (0.005%), sodium citrate (0.02%).
Preparation method：Skimmed milk powder is added in proportion first and is had warmed up into 45 DEG C of water, is stirred using blender
To abundant dissolution, then protein liquid is putted into fermenter, 0.1% Alcalase protease is added, controls temperature at 45 DEG C,
4h is kept under 80rpm, weighs glucose, yeast powder, NaCl, VC, MgSO in proportion later4·7H2O、MnSO4·H2O, lemon
Sour sodium, which is completely dissolved in suitable quantity of water, to be putted into fermenter, and adds water constant volume to mix, sterilize 20min at 115 DEG C, is cooled to 37 DEG C
Afterwards, pH to 6.0 is adjusted with sterilized NaOH.
(2) zymotechnique controls
The lactobacillus plantarum seed liquor activated is added in 10% ratio, controls pH6.0, temperature 37 with sterilized NaOH
DEG C, monitor fermentation liquid OD600It is in and stablizes to it, be determined as fermentation termination, viable count is surveyed in sampling.
(3) bacterium mud obtains and prepared by freeze-dried vaccine powder
After the fermentation liquid of fermentation ends is carried out centrifugation acquisition bacterium mud, adding suitable protective agent in proportion, (optimization is obtained
), vacuum freeze drying is carried out, controls -35 DEG C of precooling temperature, pre-freeze speed 0.6 DEG C/min, drying chamber pressure 32Pa, heating
30 DEG C of maximum temperature, vacuum freeze drying is carried out, obtains bacterium powder water content within 5%.
(4) cultivate and be lyophilized result
It the use of the lactobacillus plantarum zymotic fluid viable count that culture medium A obtains is 1.5 × 1010Cfu/mL, freeze-dried vaccine powder viable bacteria
Number is 4.0 × 1011Cfu/g, freeze-drying survival rate are 85%；
It the use of the lactobacillus plantarum zymotic fluid viable count that culture medium B is obtained is 1.0 × 1010Cfu/mL, freeze-dried vaccine powder viable bacteria
Number is 2.8 × 1011Cfu/g, freeze-drying survival rate are 75%；
It the use of the lactobacillus plantarum zymotic fluid viable count that culture medium C obtains is 1.2 × 1010Cfu/mL, freeze-dried vaccine powder viable bacteria
Number is 3.0 × 1011Cfu/g, freeze-drying survival rate are 80%；
It the use of the lactobacillus plantarum zymotic fluid viable count that culture medium D is obtained is 0.9 × 1010Cfu/mL, freeze-dried vaccine powder viable bacteria
Number is 1.5 × 1011Cfu/g, freeze-drying survival rate are 70%.
The above results show that the lactobacillus plantarum fermentation liquid ferment effect obtained using culture medium A is best, i.e., in culture medium
Contain simultaneously and resulting protein enzymatic hydrolyzate is digested by skimmed milk powder, soybean protein isolate and resulting barley is digested by pearling cone meal
When powder enzymolysis liquid, zymocyte liquid concentration highest, lactic bacteria activity is most strong, and survival rate highest is lyophilized.
Embodiment 2：Lactobacillus rhamnosus high density fermentation culture
(1) preparation of culture medium
Culture medium A：Skimmed milk powder (6%), soybean protein isolate (2%), glucose (3%), pearling cone meal (2%), yeast
Powder (1.8%), NaCl (0.23%), VC (0.3%), MgSO4·7H2O (0.02%), MnSO4·H2O (0.005%), lemon
Sour sodium (0.02%).
Preparation method：Skimmed milk powder, soybean protein isolate are added in proportion first and had warmed up into 45 DEG C of water, benefit
It is stirred with blender to abundant dissolution, then protein liquid is putted into fermenter, 0.1% Alcalase protease is added, controlled
Temperature processed keeps 4h under 45 DEG C, 80rpm, weighs glucose, yeast powder, NaCl, VC, MgSO in proportion later4·7H2O、
MnSO4·H2O, sodium citrate is completely dissolved in suitable quantity of water and puts into fermenter, and finally digests the pearling cone meal digested in advance
Liquid is added in fermentor, adds water constant volume to mix, sterilize 20min at 115 DEG C, after being cooled to 37 DEG C, with sterilized NaOH tune
Save pH to 6.0.
Culture medium B：Skimmed milk powder (6%), soybean protein isolate (2%), glucose (3%), yeast powder (1.8%),
NaCl (0.23%), VC (0.3%), MgSO4·7H2O (0.02%), MnSO4·H2O (0.005%), sodium citrate
(0.02%).
Preparation method：Weigh in proportion skimmed milk powder, soybean protein isolate, glucose, yeast powder, NaCl, VC,
MgSO4·7H2O、MnSO4·H2O, sodium citrate is completely dissolved in suitable quantity of water and puts into fermenter, and water constant volume is added to mix, in
Sterilize 20min at 115 DEG C, after being cooled to 37 DEG C, adjusts pH to 6.0 with sterilized NaOH.
Culture medium C：Skimmed milk powder (6%), soybean protein isolate (2%), glucose (3%), yeast powder (1.8%),
NaCl (0.23%), VC (0.3%), MgSO4·7H2O (0.02%), MnSO4·H2O (0.005%), sodium citrate
(0.02%).
Preparation method：Skimmed milk powder and soybean protein isolate are added in proportion first and had warmed up into 45 DEG C of water,
It is stirred using blender to abundant dissolution, then protein liquid is putted into fermenter, 0.1% Alcalase protease is added,
Control temperature keeps 4h under 45 DEG C, 80rpm, weighs glucose, yeast powder, NaCl, VC, MgSO in proportion later4·7H2O、
MnSO4·H2O, sodium citrate is completely dissolved in suitable quantity of water and puts into fermenter, and adds water constant volume to mix, sterilizes at 115 DEG C
20min after being cooled to 37 DEG C, adjusts pH to 6.0 with sterilized NaOH.
Culture medium D：Skimmed milk powder (8%), glucose (3%), yeast powder (1.8%), NaCl (0.23%), VC
(0.3%), MgSO4·7H2O (0.02%), MnSO4·H2O (0.005%), sodium citrate (0.02%).
Preparation method：Skimmed milk powder is added in proportion first and is had warmed up into 45 DEG C of water, is stirred using blender
To abundant dissolution, then protein liquid is putted into fermenter, 0.1% Alcalase protease is added, controls temperature at 45 DEG C,
4h is kept under 80rpm, weighs glucose, yeast powder, NaCl, VC, MgSO in proportion later4·7H2O、MnSO4·H2O, lemon
Sour sodium, which is completely dissolved in suitable quantity of water, to be putted into fermenter, and adds water constant volume to mix, sterilize 20min at 115 DEG C, is cooled to 37 DEG C
Afterwards, pH to 6.0 is adjusted with sterilized NaOH.
(2) zymotechnique controls
The Lactobacillus rhamnosus seed liquor activated is added in 10% ratio, controls pH6.0, temperature with sterilized NaOH
37 DEG C, monitor fermentation liquid OD600It is in and stablizes to it, be determined as fermentation termination, viable count is surveyed in sampling.
(3) bacterium mud obtains and prepared by freeze-dried vaccine powder
After the fermentation liquid of fermentation ends is carried out centrifugation acquisition bacterium mud, adding suitable protective agent in proportion, (optimization is obtained
), vacuum freeze drying is carried out, controls -35 DEG C of precooling temperature, pre-freeze speed 0.6 DEG C/min, drying chamber pressure 32Pa, heating
30 DEG C of maximum temperature, vacuum freeze drying is carried out, obtains bacterium powder water content within 5%.
(4) cultivate and be lyophilized result
It the use of the Lactobacillus rhamnosus zymotic fluid viable count that culture medium A obtains is 1.3 × 1010Cfu/mL, freeze-dried vaccine powder are living
Bacterium number is 3.0 × 1011Cfu/g, freeze-drying survival rate are 87%；
It the use of the Lactobacillus rhamnosus zymotic fluid viable count that culture medium B is obtained is 0.8 × 1010Cfu/mL, freeze-dried vaccine powder are living
Bacterium number is 1.9 × 1011Cfu/g, freeze-drying survival rate are 72%；
It the use of the Lactobacillus rhamnosus zymotic fluid viable count that culture medium C obtains is 1.0 × 1010Cfu/mL, freeze-dried vaccine powder are living
Bacterium number is 2.3 × 1011Cfu/g, freeze-drying survival rate are 80%.
It the use of the Lactobacillus rhamnosus zymotic fluid viable count that culture medium D is obtained is 0.7 × 1010Cfu/mL, freeze-dried vaccine powder are living
Bacterium number is 1.4 × 1011Cfu/g, freeze-drying survival rate are 70%.
The above results show that the Lactobacillus rhamnosus fermentation liquid ferment effect obtained using culture medium A is best, i.e. culture medium
In simultaneously containing being digested resulting protein enzymatic hydrolyzate by skimmed milk powder, soybean protein isolate and digested by pearling cone meal resulting big
When flour enzymolysis liquid, zymocyte liquid concentration highest, lactic bacteria activity is most strong, and survival rate highest is lyophilized.
The above embodiments are merely illustrative of the technical solutions of the present invention rather than limiting the scope of the invention, although ginseng
The present invention is explained in detail according to preferred embodiment, those skilled in the art should understand that, it can be to of the invention
Technical solution is modified or replaced equivalently, without departing from the spirit and scope of technical solution of the present invention.
Claims (10)
1. a kind of preparation method of lactic acid bacteria high density fermentation culture medium, which is characterized in that include the following steps：
(1) protein enzymatic hydrolyzate raw material, pearling cone meal enzymolysis liquid raw material and the basal medium raw material of following weight percent are provided,
Wherein, the raw material of the protein enzymatic hydrolyzate includes skimmed milk powder (2~10%), soybean protein isolate (1~3%)；Pearling cone meal enzyme
The raw material for solving liquid includes pearling cone meal (1~3%)；
(2) prepared by protein enzymatic hydrolyzate：Skimmed milk powder, the soybean protein isolate of the offer of step (1) are dissolved in water, it is enzyme to digest
Protein enzymatic hydrolyzate；
(3) prepared by basal medium：The basal medium raw material of the offer of step (1) is dissolved in water, obtains basal medium；
(4) prepared by pearling cone meal enzymolysis liquid：The pearling cone meal of the offer of step (1) is dissolved in water, it is enzyme to digest to obtain pearling cone meal enzymatic hydrolysis
Liquid；
(5) protein enzymatic hydrolyzate, basal medium and pearling cone meal enzymolysis liquid are mixed, constant volume, high-temperature sterilization, cooling down,
PH value is adjusted, lactic acid bacteria high density fermentation culture medium is obtained.
2. preparation method as described in claim 1, which is characterized in that in the preparation method step (1), the basis culture
Based raw material includes：Glucose (1~5%), yeast powder (1~3%), NaCl (0.1~0.3%), VC (0.2~0.4%),
MgSO4·7H2O (0.01~0.03%), MnSO4·H2O (0.003~0.007%), sodium citrate (0.01~0.03%)
3. preparation method as described in claim 1, which is characterized in that the enzyme being added in the preparation method step (2) is egg
White enzyme, additional amount are the 0.1~1% of protein enzymatic hydrolyzate raw material.
4. preparation method as described in claim 1, which is characterized in that in the preparation method step (4), the enzyme of addition is big
Wheat stickiness polysaccharide hydrolase and beta amylase, additional amount are the 0.1~1% of pearling cone meal enzymolysis liquid raw material.
5. preparation method as described in claim 1, which is characterized in that in the preparation method step (5), the culture medium
PH value is 6.0~7.0.
6. a kind of lactic acid bacteria high density fermentation culture medium, which is characterized in that the lactic acid bacteria high density fermentation culture medium is to use
Preparation method preparation gained as described in claim 1.
7. the preparation method of lactic acid bacteria high density fermentation culture medium as described in claim 1, lactic acid as claimed in claim 6
Application of the bacterium high density fermentation culture medium in lactobacillus-fermented.
8. the use as claimed in claim 7, which is characterized in that include the following steps：It will be using system as described in claim 1
Preparation Method prepares resulting lactic acid bacteria high density fermentation culture medium and is proportionally added into the lactobacillus solution activated, adjusts pH
Value, then ferments, obtains the streptococcus acidi lactici fermented solution of high density fermentation.
9. application as claimed in claim 8, which is characterized in that the additional proportion of the lactic acid bacteria high density fermentation culture medium is
The 5~20% of lactobacillus solution.
10. such as the described in any item applications of claim 7 to 9, which is characterized in that the lactic acid bacteria includes lactococcus bacterium
Kind, Lactobacillus species, Streptococcus species, Pediococcus species, Bifidobacterium strain, one in Leuconostoc strain
Kind is a variety of.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CZ308590B6 (en) * | 2019-09-25 | 2020-12-16 | Výzkumný ústav mlékárenský s.r.o. | Hypoallergenic vegan medium for cultivating lactic acid bacteria |
| CN112262888A (en) * | 2020-09-11 | 2021-01-26 | 云南皇氏来思尔乳业有限公司 | Bacterial enzyme synergistic starter for preparing yoghourt |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1353191A (en) * | 2000-11-09 | 2002-06-12 | 火箭兄弟公司 | Method of preparing fermentation culture medium from renewable raw material |
| CN103445068A (en) * | 2013-06-26 | 2013-12-18 | 江苏大学 | Preparation method of barley extract fermented by lactobacillus and anti-tumor effect of barley extract fermented by lactobacillus |
-
2018
- 2018-07-09 CN CN201810745867.8A patent/CN108841753A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1353191A (en) * | 2000-11-09 | 2002-06-12 | 火箭兄弟公司 | Method of preparing fermentation culture medium from renewable raw material |
| US20020079268A1 (en) * | 2000-11-09 | 2002-06-27 | Jean-Jacques Caboche | Process for preparing a fermentation medium from a renewable raw material |
| CN103445068A (en) * | 2013-06-26 | 2013-12-18 | 江苏大学 | Preparation method of barley extract fermented by lactobacillus and anti-tumor effect of barley extract fermented by lactobacillus |
Non-Patent Citations (1)
| Title |
|---|
| 白凤翎: "蛋白水解物促乳酸菌增殖及高密度培养体系研究", 《中国博士学位论文全文数据库》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CZ308590B6 (en) * | 2019-09-25 | 2020-12-16 | Výzkumný ústav mlékárenský s.r.o. | Hypoallergenic vegan medium for cultivating lactic acid bacteria |
| CN112262888A (en) * | 2020-09-11 | 2021-01-26 | 云南皇氏来思尔乳业有限公司 | Bacterial enzyme synergistic starter for preparing yoghourt |
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