CN107858320A - Bacteria culture media, cultural method and the application of the two - Google Patents
Bacteria culture media, cultural method and the application of the two Download PDFInfo
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- CN107858320A CN107858320A CN201711245493.5A CN201711245493A CN107858320A CN 107858320 A CN107858320 A CN 107858320A CN 201711245493 A CN201711245493 A CN 201711245493A CN 107858320 A CN107858320 A CN 107858320A
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Abstract
The present invention relates to strain culturing technical field, specifically, there is provided a kind of bacteria culture media, cultural method and the application of the two.Bacteria culture media provided by the invention, including basal medium and additive, additive include L cysteines and sodium thioglycolate.The culture medium can provide nitrogen source, vitamin, growth factor and fermentable saccharide needed for lactic acid bacteria growth etc., the growth of other miscellaneous bacterias can be suppressed simultaneously, the addition of L cysteines and thioglycolic acid sodium additives can slow down the dissolving of lactic acid bacterial cell wall, so as to reduce the dead bacterium number in fermentation later stage, lactic acid bacteria can keep possessing high activity while high-density growth, can promote the fast-growth of lactic acid bacteria and a large amount of amplifications.Lactic acid bacteria cultural method provided by the invention, it is simple efficiently, cost is low, can largely expand the highdensity lactic acid bacteria of high activity in short cycle, the industrialization culture to lactic acid bacteria is most important.
Description
Technical field
The present invention relates to strain culturing technical field, in particular to a kind of bacteria culture media, cultural method and the two
Application.
Background technology
Probiotics especially lactic acid bacteria plays an important roll in microecological balance, preventing and treating infection is safeguarded.Saliva lactic acid bar
Bacterium is common lactic acid bacteria in daily life, and it has wide antimicrobial spectrum, to Escherichia coli, staphylococcus aureus, pneumonia
Klebsiella, Shigella, streptococcus pneumonia etc. all have good inhibition, in growth course, can secrete peroxide
Change the materials such as hydrogen, hydrogen sulfide and antibacterial peptide, directly suppress or kill harmful bacteria growing；Ring is reduced by secreting the organic acids such as lactic acid
Border pH value, reduce pathogenic bacteria etc. and grow；It has a major impact to enteric microorganism simultaneously, no matter in food fermentation, health products, life
Thing medicine and other fields suffer from being widely applied.
Probiotic products large-scale commercial in recent years, industrial fermentation production scale is promoted to expand rapidly, industrial fermentation is most
The lifting of whole viable count, it is the target that every probiotics manufacturing enterprise is pursued, this requires large scale fermentation engineering to have a height
Imitate fermentation pattern.Therefore, the technology and technique for studying large-scale industrial fermentation engineering bacteria seem extremely important.Lactic acid bacteria
The preparation of high density fermentation agent is realized to its high activity, the key of highdensity Multiplying culture.
In view of this, it is special to propose the present invention.
The content of the invention
The first object of the present invention is to provide a kind of bacteria culture media, and the second object of the present invention is to provide a kind of breast
Acetic bacterial cultural method, the of the invention the 3rd at present in providing above-mentioned culture medium or cultural method in lactic acid bacteria is cultivated
Application, with alleviate lactic acid bacteria culture units yield and activity are low in the prior art, it is impossible to the technology of proliferation of high-density culture
Problem.
In order to realize the above-mentioned purpose of the present invention, spy uses following technical scheme：
The invention provides a kind of bacteria culture media, the bacteria culture media includes basal medium and additive, described
Additive includes Cys and sodium thioglycolate.
Further, the basal medium includes the raw material of following parts by weight：Glucose 5-20 parts, peptone 0.5-
2.0 parts, beef extract 0.1-1.0 parts, yeast extract 0.1-1.0 parts, dipotassium hydrogen phosphate 0.1-0.5 parts, triammonium citrate 0.1-
0.5 part, sodium acetate 0.1-0.5 parts, magnesium sulfate 0.01-0.08 parts, manganese sulfate 0.001-0.007 parts and Tween-80 0.05-0.3
Part.
Further, the additive includes the raw material of following parts by weight：Cys 0.01-0.03 parts and thio second
Alkyd sodium 0.002-0.007 parts.
Further, the additive includes the raw material of following parts by weight：Cys 0.01-0.02 parts and thio second
Alkyd sodium 0.002-0.004 parts.
Further, each raw material is prepared by formula ratio adds pure water to be dissolved after supplying 100 parts, adjusts pH to 6.2-6.6,
The bacteria culture media is obtained after sterilizing.
Further, the culture medium solidifying agent that parts by weight are 2.3-2.7 parts is also included when preparing solid medium.
PH comprises the following steps present invention also offers a kind of lactic acid bacteria cultural method：Regulation culture temperature is 36.5-
37.5 DEG C, using above-mentioned bacteria culture media, by lactic acid bacteria actication of culture, slant strains are then prepared, then carry out one-level hair
Ferment and second order fermentation, produce acid to zymotic fluid and stop, finally giving high-density lactic acid bacteria zymotic fluid.
Further, the pressure in the fermentation tank of the second order fermentation is 0.04-0.06MPa；
Preferably, nitrogen is passed through in the fermentation tank of the second order fermentation, it is 0.04-0.06MPa to make pressure；
Preferably, the nitrogen is High Purity Nitrogen.
Further, it is 5.8-6.2 to regulate and control pH value.
Present invention also offers above-mentioned bacteria culture media or above-mentioned lactic acid bacteria cultural method in culture lactic acid bacteria
In application.
Compared with prior art, beneficial effects of the present invention are：
Bacteria culture media provided by the invention, including basal medium and additive, additive include Cys and
Sodium thioglycolate.The culture medium can provide nitrogen source, vitamin, growth factor and the fermentable sugars needed for lactic acid bacteria growth
Class etc., while the growth of other miscellaneous bacterias can be suppressed, the addition of Cys and thioglycolic acid sodium additives can slow down
The dissolving of lactic acid bacterial cell wall, so as to reduce the dead bacterium number in fermentation later stage, lactic acid bacteria can keep high-density growth
Possess high activity simultaneously, the fast-growth of lactic acid bacteria and a large amount of amplifications can be promoted.Lactic acid bacteria culture provided by the invention
Method, it is simple efficiently, cost is low, can largely expand the highdensity lactic acid bacteria of high activity in short cycle, to the work of lactic acid bacteria
Industry culture is most important.
Embodiment
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the present invention.It is unreceipted specific in embodiment
Condition person, the condition suggested according to normal condition or manufacturer are carried out.
Bacteria culture media provided by the invention, including basal medium and additive, additive include Cys and
Sodium thioglycolate.
The culture medium can provide nitrogen source, vitamin, growth factor and fermentable saccharide needed for lactic acid bacteria growth etc.,
The growths of other miscellaneous bacterias can be suppressed simultaneously, it is thin that the additions of Cys and thioglycolic acid sodium additives can slow down lactic acid
The dissolving of bacterium cell membrane, so as to reduce the dead bacterium number in fermentation later stage, lactic acid bacteria can keep gathering around while high-density growth
There is high activity, the fast-growth of lactic acid bacteria and a large amount of amplifications can be promoted.
It is preferably carried out at one in mode, basal medium includes the raw material of following parts by weight：Glucose 5-20 parts, egg
White peptone 0.5-2.0 parts, beef extract 0.1-1.0 parts, yeast extract 0.1-1.0 parts, dipotassium hydrogen phosphate 0.1-0.5 parts, citric acid
Triamine 0.1-0.5 parts, sodium acetate 0.1-0.5 parts, magnesium sulfate 0.01-0.08 parts, manganese sulfate 0.001-0.007 parts and Tween-80
0.05-0.3 parts.
Peptone, beef extract and yeast extract provide nitrogen source, vitamin and growth factor；Glucose is fermentable sugars
Class；Dipotassium hydrogen phosphate is acid-base buffer agent；Triammonium citrate, sodium acetate, magnesium sulfate, manganese sulfate and Tween-80 provide for bacterium
Growth factor, other miscellaneous bacterias can also be suppressed.
It should be noted that glucose parts by weight are typical but non-limiting is 5 parts, 6 parts, 7 parts, 8 parts, 9 parts, 10
Part, 11 parts, 12 parts, 13 parts, 14 parts, 15 parts, 16 parts, 17 parts, 18 parts, 19 parts or 20 parts；Peptone parts by weight are typical but non-
It is restricted for 0.5 part, 0.6 part, 0.7 part, 0.8 part, 0.9 part, 1.0 parts, 1.1 parts, 1.2 parts, 1.3 parts, 1.4 parts, 1.5 parts,
1.6 parts, 1.7 parts, 1.8 parts, 1.9 parts or 2.0 parts；It is 0.1 part that beef extract parts by weight are typical but non-limiting, 0.2 part,
0.3 part, 0.4 part, 0.5 part, 0.6 part, 0.7 part, 0.8 part, 0.9 part or 1.0 parts；Yeast extract parts by weight are typical but unrestricted
Property for 0.1 part, 0.2 part, 0.3 part, 0.4 part, 0.5 part, 0.6 part, 0.7 part, 0.8 part, 0.9 part or 1.0 parts；Dipotassium hydrogen phosphate
Typical but non-limiting parts by weight are 0.1 part, 0.2 part, 0.3 part, 0.4 part or 0.5 part；Triammonium citrate parts by weight allusion quotation
Type but it is nonrestrictive be 0.1 part, 0.2 part, 0.3 part, 0.4 part or 0.5 part；Sodium acetate parts by weight are typical but non-limiting
For 0.1 part, 0.2 part, 0.3 part, 0.4 part or 0.5 part；It is 0.01 part that magnesium sulfate parts by weight are typical but non-limiting, 0.02
Part, 0.03 part, 0.04 part, 0.05 part, 0.06 part, 0.07 part or 0.08 part；Manganese sulfate parts by weight are typical but non-limiting
For 0.01 part, 0.02 part, 0.03 part, 0.04 part, 0.05 part, 0.06 part or 0.07 part；Tween-80 parts by weight are typical but non-limit
Property processed for 0.05 part, 0.06 part, 0.07 part, 0.08 part, 0.09 part, 0.1 part, 0.2 part or 0.3 part.
In a preferred embodiment, basal medium includes the raw material of following parts by weight：Glucose 10-20
Part, peptone 1.0-2.0 parts, beef extract 0.5-1.0 parts, yeast extract 0.5-1.0 parts, dipotassium hydrogen phosphate 0.2-0.5 parts, lemon
Lemon acid triamine 0.2-0.5 parts, sodium acetate 0.2-0.5 parts, magnesium sulfate 0.05-0.08 parts and are told at manganese sulfate 0.005-0.007 parts
Warm -80 0.1-0.3 parts.
In a preferred embodiment, basal medium includes the raw material of following parts by weight：10 parts of glucose, egg
White 1.0 parts of peptone, 0.5 part of beef extract, 0.5 part of yeast extract, 0.2 part of dipotassium hydrogen phosphate, 0.2 part of triammonium citrate, sodium acetate
0.2 part, 0.05 part of magnesium sulfate, 0.1 part of 0.005 part of manganese sulfate and Tween-80.
In one preferred embodiment, additive includes the raw material of following parts by weight：Cys 0.01-0.03
Part and sodium thioglycolate 0.002-0.007 parts.
It should be noted that it is 0.01 part, 0.02 part or 0.03 that Cys parts by weight are typical but non-limiting
Part；It is 0.002 part that sodium thioglycolate parts by weight are typical but non-limiting, 0.003 part, 0.004 part, 0.005 part,
0.006 part or 0.007 part.
In a preferred embodiment, additive includes the raw material of following parts by weight：Cys 0.01-
0.02 part and sodium thioglycolate 0.002-0.004 parts.
In a preferred embodiment, additive includes the raw material of following parts by weight：0.02 part of Cys
With 0.003 part of sodium thioglycolate.
Addition Cys and sodium thioglycolate reducing agent can reduce the initial oxidation reduction potential of nutrient solution rapidly,
Be advantageous to the fast breeding of thalline.
In one preferred embodiment, each raw material is prepared by formula ratio, adds pure water to be dissolved after supplying 100 parts,
PH to 6.2-6.6 is adjusted, above-mentioned bacteria culture media is obtained after sterilizing.
In one preferred embodiment, the culture that parts by weight are 2.3-2.7 parts is also included when preparing solid medium
Base coagulator.Culture medium solidifying agent can be agar, gelatin, inorganic silica gel or carragheen, preferably agar.There is provided in the present invention
Fluid nutrient medium in add culture medium solidifying agent, obtain solid medium.The parts by weight of culture medium solidifying agent are typical but non-limit
Property processed for 2.3 parts, 2.4 parts, 2.5 parts, 2.6 parts or 2.7 parts.
Present invention also offers a kind of lactic acid bacteria cultural method, comprise the following steps：Regulation culture temperature is 36.5-
37.5 DEG C, cultivated using above-mentioned optimization lactic acid bacteria, by lactic acid bacteria actication of culture, then prepare slant strains, then enter
Row one grade fermemtation and second order fermentation, produce acid to zymotic fluid and stop, finally giving high-density lactic acid bacteria zymotic fluid.Need what is illustrated
To be that cultivation temperature is typical but non-limiting be 36.5 DEG C, 36.6 DEG C, 36.7 DEG C, 36.8 DEG C, 36.9 DEG C, 37.0 DEG C, 37.1
DEG C, 37.2 DEG C, 37.3 DEG C, 37.4 DEG C or 37.5 DEG C.Lactic acid bacteria cultural method provided by the invention, it is simple efficiently, cost it is low,
The highdensity lactic acid bacteria of high activity can be largely expanded in the short cycle, the industrialization culture to lactic acid bacteria is most important.
It is preferably carried out at one in mode, the pressure in the fermentation tank of second order fermentation is 0.04-0.06MPa；
Preferably, nitrogen is passed through in the fermentation tank of the second order fermentation, it is 0.04-0.06MPa to make pressure；
Preferably, the nitrogen is High Purity Nitrogen.
The anaerobic environment that lactic acid bacteria growth needs is provided, while suppresses the growth of miscellaneous bacteria.Pressure is typical but non-limiting
For 0.04MPa, 0.05MPa or 0.06MPa.
It is preferably carried out at one in mode, regulation and control pH value is 5.8-6.2, and pH adjusting agent is that acetic acid parts by weight are 9-18
The acetic acid solution and NaOH parts by weight of part are the NaOH aqueous slkalis of 10-20 parts.It is 5.8 that pH value is typical but non-limiting,
5.9th, 6.0,6.1 or 6.2.
Present invention also offers above-mentioned bacteria culture media or above-mentioned lactic acid bacteria cultural method in culture lactic acid bacteria
In application.
In order to help to further understand the present invention, technical scheme is carried out in conjunction with preferred embodiment detailed
Explanation.
Embodiment 1
A kind of bacteria culture media, including：5 parts of glucose, 0.5 part of peptone, 0.1 part of beef extract, yeast extract 0.1
Part, 0.1 part of dipotassium hydrogen phosphate, 0.1 part of triammonium citrate, 0.1 part of sodium acetate, 0.01 part of magnesium sulfate, 0.001 part of manganese sulfate, tell
0.002 part of -80 0.05 parts of temperature, 0.01 part of Cys and sodium thioglycolate；
100 parts are supplied with pure water.
Embodiment 2
A kind of bacteria culture media, including：20 parts of glucose, 2.0 parts of peptone, 1.0 parts of beef extract, yeast extract 1.0
Part, 0.5 part of dipotassium hydrogen phosphate, 0.5 part of triammonium citrate, 0.5 part of sodium acetate, 0.08 part of magnesium sulfate, 0.007 part of manganese sulfate, tell
0.007 part of -80 0.3 parts of temperature, 0.03 part of Cys and sodium thioglycolate；
100 parts are supplied with pure water.
Embodiment 3
A kind of bacteria culture media, including：10 parts of glucose, 1.0 parts of peptone, 0.5 part of beef extract, yeast extract 0.5
Part, 0.2 part of dipotassium hydrogen phosphate, 0.2 part of triammonium citrate, 0.2 part of sodium acetate, 0.05 part of magnesium sulfate, 0.005 part of manganese sulfate, tell
0.003 part of -80 0.1 parts of temperature, 0.02 part of Cys and sodium thioglycolate；
100 parts are supplied with pure water.
Embodiment 4
A kind of bacteria culture media, including：5 parts of glucose, 0.5 part of peptone, 0.1 part of beef extract, yeast extract 0.1
Part, 0.1 part of dipotassium hydrogen phosphate, 0.1 part of triammonium citrate, 0.1 part of sodium acetate, 0.01 part of magnesium sulfate, 0.001 part of manganese sulfate, tell
0.002 part of -80 0.05 parts of temperature, 0.01 part of Cys and sodium thioglycolate；
2.3 parts of agar is added, 100 parts are supplied with pure water.
Embodiment 5
A kind of bacteria culture media, including：20 parts of glucose, 2.0 parts of peptone, 1.0 parts of beef extract, yeast extract 1.0
Part, 0.5 part of dipotassium hydrogen phosphate, 0.5 part of triammonium citrate, 0.5 part of sodium acetate, 0.08 part of magnesium sulfate, 0.007 part of manganese sulfate, tell
0.007 part of -80 0.3 parts of temperature, 0.03 part of Cys and sodium thioglycolate；
2.7 parts of agar is added, 100 parts are supplied with pure water.
Embodiment 6
A kind of bacteria culture media, including：10 parts of glucose, 1.0 parts of peptone, 0.5 part of beef extract, yeast extract 0.5
Part, 0.2 part of dipotassium hydrogen phosphate, 0.2 part of triammonium citrate, 0.2 part of sodium acetate, 0.05 part of magnesium sulfate, 0.005 part of manganese sulfate, tell
0.003 part of -80 0.1 parts of temperature, 0.02 part of Cys and sodium thioglycolate；
2.5 parts of agar is added, 100 parts are supplied with pure water.
Embodiment 7-9
Experiment Lactobacillus salivarius, a kind of probiotics of Healthy People fecal sample is located away from, there is good acidproof, resistance to courage
The abilities such as juice, the bacterial strain were preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms on December 26th, 2012
The heart, preserving number are CGMCC No7045.
A kind of lactic acid bacteria cultural method, comprises the following steps：
(a) activation of strain：Lactobacillus salivarius cryopreservation tube of the freezen protective in -40 DEG C is opened, is inoculated in embodiment 4-6
Solid plate culture medium on, temperature be 36.5 DEG C under the conditions of cultivate 18 hours, so passage the generation of activation culture 2 activated
Strain；
(b) slant strains make：The single bacterium colony of activation in (a) is taken, is inoculated in any one of embodiment 4-6 solid slope training
Support in base, temperature is 36.5 DEG C, is cultivated 16 hours under anaerobic condition；
(c) primary seed solution is prepared：(b) is made into cultured inclined plane inoculating into embodiment 1-3 bacteria culture media,
37 DEG C are cultivated 12 hours；
(d) secondary seed solution is prepared：Primary seed solution in (c) is inoculated into containing embodiment according to 5% inoculum concentration
The seeding tank fermented and cultured of 1-3 culture medium, 37 DEG C of temperature control, pH maintain 5.8, in tank with 99.99% N2Pressurize
0.04MPa, cultivate 10 hours；
(e) fermented and cultured：(d) secondary seed solution is inoculated into containing embodiment according to 5% inoculum concentration by pressure differential method
In the fermentation tank culture medium of 1-3 culture medium, 37 DEG C of temperature control, pH maintains 5.8, in tank with 99.99% N2Pressurize
0.04 culture, when fermentation and acid stops, NaOH aqueous slkalis do not stop fermentation in the stream added-time, obtain highdensity Lactobacillus salivarius training
Nutrient solution.
Embodiment 10-12
Experiment Lactobacillus salivarius, a kind of probiotics of Healthy People fecal sample is located away from, there is good acidproof, resistance to courage
The abilities such as juice, the bacterial strain were preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms on December 26th, 2012
The heart, preserving number are CGMCC No7045.
A kind of lactic acid bacteria cultural method, comprises the following steps：
(a) activation of strain：Lactobacillus salivarius cryopreservation tube of the freezen protective in -40 DEG C is opened, is inoculated in embodiment 4-6
Solid plate culture medium on, temperature be 37 DEG C under the conditions of cultivate 20 hours, so pass on the generation of activation culture 2 obtain activation bacterium
Kind；
(b) slant strains make：The single bacterium colony of activation in (a) is taken, is inoculated in any one of embodiment 4-6 solid slope training
Support in base, temperature is 37 DEG C, is cultivated 18 hours under anaerobic condition；
(c) primary seed solution is prepared：(b) is made into cultured inclined plane inoculating into embodiment 1-3 bacteria culture media,
37.5 DEG C are cultivated 10 hours；
(d) secondary seed solution is prepared：Primary seed solution in (c) is inoculated into containing embodiment according to 5% inoculum concentration
The seeding tank fermented and cultured of 1-3 culture medium, 37.5 DEG C of temperature control, pH maintain 6.2, in tank with 99.99% N2Pressurize
0.06MPa, cultivate 8 hours；
(e) fermented and cultured：(d) secondary seed solution is inoculated into containing embodiment according to 5% inoculum concentration by pressure differential method
In the fermentation tank culture medium of 1-3 culture medium, 37.5 DEG C of temperature control, pH maintains 6.2, in tank with 99.99% N2Pressurize
0.06MPa is cultivated, and when fermentation and acid stops, NaOH aqueous slkalis do not stop fermentation in the stream added-time, obtain highdensity saliva breast bar
Bacteria culture fluid.
Comparative example 1
A kind of bacteria culture media, in parts by weight meter include：10 parts of glucose, 1.0 parts of peptone, 0.5 part of beef extract,
0.5 part of yeast extract, 0.2 part of dipotassium hydrogen phosphate, 0.2 part of triammonium citrate, 0.2 part of sodium acetate, 0.05 part of magnesium sulfate, manganese sulfate
0.005 part and 0.1 part of Tween-80；
100 parts are supplied with pure water.
Comparative example 2
A kind of bacteria culture media, including：10 parts of glucose, 1.0 parts of peptone, 0.5 part of beef extract, yeast extract 0.5
Part, 0.2 part of dipotassium hydrogen phosphate, 0.2 part of triammonium citrate, 0.2 part of sodium acetate, 0.05 part of magnesium sulfate, 0.005 part of manganese sulfate, tell
- 80 0.1 parts of temperature, 0.002 part of sodium thioglycolate；
100 parts are supplied with pure water.
Comparative example 3
A kind of bacteria culture media, including：10 parts of glucose, 1.0 parts of peptone, 0.5 part of beef extract, yeast extract 0.5
Part, 0.2 part of dipotassium hydrogen phosphate, 0.2 part of triammonium citrate, 0.2 part of sodium acetate, 0.05 part of magnesium sulfate, 0.005 part of manganese sulfate, tell
- 80 0.1 parts of temperature, 0.007 part of Cys；
100 parts are supplied with pure water.
Comparative example 4
A kind of bacteria culture media, including：20 parts of glucose, 2.0 parts of peptone, 1.0 parts of beef extract, yeast extract 1.0
Part, 0.5 part of dipotassium hydrogen phosphate, 0.5 part of triammonium citrate, 0.5 part of sodium acetate, 0.08 part of magnesium sulfate, 0.007 part of manganese sulfate, tell
0.010 part of -80 0.3 parts of temperature, 0.08 part of Cys and sodium thioglycolate；
100 parts are supplied with pure water.
Comparative example 5
A kind of bacteria culture media, including：25 parts of glucose, 2.2 parts of peptone, 1.4 parts of beef extract, yeast extract 1.6
Part, 0.3 part of dipotassium hydrogen phosphate, 0.8 part of triammonium citrate, 0.7 part of sodium acetate, 0.10 part of magnesium sulfate, 0.009 part of manganese sulfate, tell
0.010 part of -80 0.5 parts of temperature, 0.08 part of Cys and sodium thioglycolate；
100 parts are supplied with pure water.
Test example
Lyophilized bacterium powder will be prepared into identical method with the Lactobacillus salivarius of embodiment 1-12 and comparative example 1-5 cultures,
Detect the number of viable of bacterium powder.As a result it is as shown in the table.
| Viable count | |
| Embodiment 1 | 1.8×1011cfu/g |
| Embodiment 2 | 1.6×1011cfu/g |
| Embodiment 3 | 2.1×1011cfu/g |
| Embodiment 4 | 1.7×1011cfu/g |
| Embodiment 5 | 1.6×1011cfu/g |
| Embodiment 6 | 1.3×1011cfu/g |
| Embodiment 7 | 2.0×1011cfu/g |
| Embodiment 8 | 2.6×1011cfu/g |
| Embodiment 9 | 3.5×1011cfu/g |
| Embodiment 10 | 2.2×1011cfu/g |
| Embodiment 11 | 2.5×1011cfu/g |
| Embodiment 12 | 3.0×1011cfu/g |
| Comparative example 1 | 1.0×108cfu/g |
| Comparative example 2 | 1.2×109cfu/g |
| Comparative example 3 | 1.6×109cfu/g |
| Comparative example 4 | 2.3×1010cfu/g |
| Comparative example 5 | 1.2×1010cfu/g |
From experimental result above, the bacteria culture media that embodiment 1-6 is provided can effectively improve lactic acid bacteria
Activity；Embodiment 7-12 viable count is higher than embodiment 1-6, and it is highly dense to illustrate that cultural method provided by the invention can be realized
Degree, the culture lactic acid bacteria of high activity；Additive Cys and thioglycolic are can be seen that from comparative example 1-5 viable count
Decisive role is played in high density of the sour sodium all to lactic acid bacteria, high activity culture.
Although illustrate and describing the present invention with specific embodiment, but will be appreciated that without departing substantially from the present invention's
Many other change and modification can be made in the case of spirit and scope.It is, therefore, intended that in the following claims
Including belonging to all such changes and modifications in the scope of the invention.
Claims (10)
1. a kind of bacteria culture media, it is characterised in that the bacteria culture media includes basal medium and additive, the addition
Agent includes Cys and sodium thioglycolate.
2. bacteria culture media according to claim 1, it is characterised in that the basal medium includes following parts by weight
Raw material：Glucose 5-20 parts, peptone 0.5-2.0 parts, beef extract 0.1-1.0 parts, yeast extract 0.1-1.0 parts, phosphoric acid hydrogen
Dipotassium 0.1-0.5 parts, triammonium citrate 0.1-0.5 parts, sodium acetate 0.1-0.5 parts, magnesium sulfate 0.01-0.08 parts, manganese sulfate
0.001-0.007 parts and Tween-80 0.05-0.3 parts.
3. bacteria culture media according to claim 1, it is characterised in that the additive includes the original of following parts by weight
Material：Cys 0.01-0.03 parts and sodium thioglycolate 0.002-0.007 parts.
4. bacteria culture media according to claim 3, it is characterised in that the additive includes the original of following parts by weight
Material：Cys 0.01-0.02 parts and sodium thioglycolate 0.002-0.004 parts.
5. according to the bacteria culture media described in claim any one of 1-4, it is characterised in that each raw material is prepared by formula ratio and is added
Pure water is dissolved after supplying 100 parts, adjusts pH to 6.2-6.6, and the bacteria culture media is obtained after sterilizing.
6. bacteria culture media according to claim 5, it is characterised in that be also including parts by weight when preparing solid medium
The culture medium solidifying agent of 2.3-2.7 parts.
7. a kind of lactic acid bacteria cultural method, it is characterised in that comprise the following steps：Regulation culture temperature is 36.5-37.5 DEG C,
Using the bacteria culture media described in claim any one of 1-6, by lactic acid bacteria actication of culture, by the lactic acid bacteria system after activation
Standby slant strains, and one grade fermemtation and second order fermentation are carried out, producing acid to zymotic fluid stops, and obtains high-density lactic acid bacteria fermentation
Liquid.
8. cultural method according to claim 7, it is characterised in that the pressure in the fermentation tank of the second order fermentation is
0.04-0.06MPa；
Preferably, nitrogen is passed through in the fermentation tank of the second order fermentation, it is 0.04-0.06MPa to make pressure；
Preferably, the nitrogen is High Purity Nitrogen.
9. cultural method according to claim 8, it is characterised in that regulation and control pH value is 5.8-6.2.
10. the lactic acid bacteria training as described in the bacteria culture media or claim any one of 7-9 as described in claim any one of 1-6
Application of the method for supporting in lactic acid bacteria is cultivated.
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| CN110066734A (en) * | 2019-05-20 | 2019-07-30 | 广州天适立农生态农业发展有限公司 | A kind of quickly breeding biological tropina culture medium |
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