CN108315277A - One lactobacillus plantarum and a kind of method of freeze-drying lactic acid bacteria - Google Patents
One lactobacillus plantarum and a kind of method of freeze-drying lactic acid bacteria Download PDFInfo
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Abstract
This application discloses a lactobacillus plantarum LP5, a kind of method being freeze-dried lactic acid bacteria and the lactobacillus plantarum LP5 bacterium powders prepared by the method.The method to thalline before carrying out vacuum freeze drying, upper machine liquid is set to cool down rapidly using liquid nitrogen, to reduce cellular damage of lactic acid bacteria thalline during vacuum refrigeration to greatest extent, storage-stable of the lactic acid bacteria under normal temperature condition can be significantly improved, later stage vacuum drying directly carries out radiant temperature drying, and the energy is more saved compared with conventional method.Lactobacillus plantarum (the Lactobacillus plantarum LP5) bacterium powder, the storage-stable under normal temperature condition significantly improve, preservation 360 days under 25 DEG C of constant temperatures, and Viable detection is 75.0% or more.
Description
Technical field
The invention belongs to probiotics fields, and in particular to a lactobacillus plantarum and a kind of freeze-drying lactic acid bacteria
Method.
Background technology
Early in the seventies, some American-European developed countries develop novel probiotics in succession, it is desirable to a certain degree
Upper substitute antibiotics, and play the effect for adjusting body immunity.In recent years, probiotics were recognized with China consumer
Continuous intensification, lactobacillus micro-ecological system with its recuperating gastrointestinal tract and improve immunity in terms of significant advantage, increasingly by
The concern of people.
Lactobacillus micro-ecological preparation generally goes through vacuum freeze drying processing before launch.But in actual production
In the process, traditional Vacuum Freezing ＆ Drying Technology, it is more slow in freezing initial stage temperature-fall period, due to being deposited in outside environment
In multi-solvents and inorganic salts, wherein the low solvent of freezing point first crystallizes, and is crystallized after the low solvent of freezing point, and the nothing in system
Machine salt does not crystallize, this results in the liquid concentration of outside environment increasing, is gradually converted into glassy state, so as to cause thin
The osmotic pressure of born of the same parents' external environment gradually increases, and then intracellular cytoplasm is caused to change, also, iuntercellular and intracellular
The random arrangement of ice crystal, intercellular pressure acts directly on cytoplasma membrane, change cytoplasma membrane mobility and
Other physics and chemical property, it is also possible to puncture cell membrane.These combined factors cause cell membrane can be by oxidative damage so that
Lactic acid bacteria itself is poor to the resistance of external environment, and bioactivity is gradually lost, so as to cause lactic acid bacteria solid food shelf life
It is shorter, the serious producing and selling and application for restricting lactic acid bacteria series of products.
Therefore, it is extremely urgent to seek a kind of processing technology that storage-stable under lactic acid bacteria normal temperature condition can be improved.
Invention content
The first purpose of the application is to provide lactobacillus plantarum LP5 (the Lactobacillus plantarum of one plant of separation
LP5, L.plantarum LP5).
In order to achieve the above objectives, the application is achieved through the following technical solutions：
Lactobacillus plantarum (Lactobacillus plantarum LP5, L.plantarum LP5) sieves by the following method
Choosing obtains：
Lactobacillus plantarum LP5 is isolated from the pickles sample of spontaneous fermentation, passes through MRS solid mediums (DifcoTM
Lactobacilli MRS Agar) scribing line, in 37 DEG C, constant temperature incubation 48h is isolated and purified.It identifies and ties through Physiology and biochemistry
Conjunction 16s rRNA Molecular Identifications are 1 lactobacillus plantarum, are named as lactobacillus plantarum LP5 (Lactobacillus plantarum
LP5), which is preserved in China Committee for Culture Collection of Microorganisms's General Microbiological Culture on 25th in September in 2017
Collection.
The mechanism that the bacterial strain submits patent to approve is carried out preservation by the application, and microbial preservation number is：CGMCC
No.13459；Classification And Nomenclature is：Lactobacillus plantarum (Lactobacillus plantarum)；Depositary institution：China Microbiological bacterium
Kind preservation administration committee General Microbiological Culture collection (State Patent Office specifies patent Organism Depositary) preservation；
The preservation time：On September 25th, 2017；Preservation address：City of BeiJing, China Chaoyang District North Star West Road 1 institute 3, the Chinese Academy of Sciences is micro-
Biological study institute.
The separated lactobacillus plantarum strain of the application (Lactobacillus plantarum LP5) has following biology
Learn characteristic：Strain is gram sample bacterium, is in direct rod shape, individually, sometimes in pairs or at chain.Bacterium colony on MRS solid mediums
Diameter 1-3mm, milky, protrusion, rounded, surface is smooth.Optimum growth temperature is 25~37 DEG C, anaerobism or amphimicrobian,
Optimal pH 6.5 or so is grown muddy in MRS fluid nutrient mediums.
The second purpose of the application is to provide a kind of method of freeze-drying lactic acid bacteria, the method includes：
Step 1, lactic acid bacteria bacterium mud is mixed with water, small molecular sugar, protein and oxygen scavenger, is stirred, upper machine liquid is made；
Step 2, upper machine liquid temperature in 10min prepared by step 1 is made to be down to -50 DEG C, heat preservation；
Step 3, system prepared by step 2 is placed in -20 DEG C of environment, is frozen under vacuum according to temperature programming
It is dry.
It is freeze-dried that the third purpose of the application is to provide a kind of lactobacillus plantarum prepared by the method, the plant breast
The freeze-dried middle viable count of lactobacillus of bacillus is 4.6 × 1011CFU/g or more, preservation 360 days under 25 DEG C of constant temperatures, viable bacteria is deposited
Motility rate is 75.0% or more.
The method of freeze-drying lactic acid bacteria provided by the present application is markedly different from partition board Refrigeration Technique traditional at present, cooling
Cellular damage of lactic acid bacteria thalline during vacuum refrigeration is reduced rapidly and to greatest extent, lactic acid bacteria can be significantly improved normal
Storage-stable under the conditions of temperature, later stage vacuum drying directly carry out radiant temperature drying, the energy are more saved compared with conventional method.
The lactic acid bacteria freeze drying powder prepared using method provided by the present application, the plant prepared especially with the application method
Lactobacillus (Lactobacillus plantarum LP5) bacterium powder, the storage-stable under normal temperature condition significantly improves, 25
Preservation 360 days under DEG C constant temperature, Viable detection are 75.0% or more.
Specific implementation mode
Lactobacillus plantarum (Lactobacillus plantarum LP5, L.plantarum LP5) sieves by the following method
Choosing obtains：
Lactobacillus plantarum LP5 is isolated from the pickles sample of spontaneous fermentation, passes through MRS solid mediums (DifcoTM
Lactobacilli MRS Agar) scribing line, in 37 DEG C, constant temperature incubation 48h is isolated and purified.It identifies and ties through Physiology and biochemistry
Conjunction 16s rRNA Molecular Identifications are 1 lactobacillus plantarum, are named as lactobacillus plantarum LP5 (Lactobacillus plantarum
LP5), which is preserved in China Committee for Culture Collection of Microorganisms's General Microbiological Culture on 25th in September in 2017
Collection.
The mechanism that the bacterial strain submits patent to approve is carried out preservation by the application, and microbial preservation number is：CGMCC
No.13459；Classification And Nomenclature is：Lactobacillus plantarum (Lactobacillus plantarum)；Depositary institution：China Microbiological bacterium
Kind preservation administration committee General Microbiological Culture collection (State Patent Office specifies patent Organism Depositary) preservation；
The preservation time：On September 25th, 2017；Preservation address：City of BeiJing, China Chaoyang District North Star West Road 1 institute 3, the Chinese Academy of Sciences is micro-
Biological study institute.
The separated lactobacillus plantarum strain of the application (Lactobacillus plantarum LP5) has following biology
Learn characteristic：Strain is gram sample bacterium, is in direct rod shape, individually, sometimes in pairs or at chain.Bacterium colony on MRS solid mediums
Diameter 1-3mm, milky, protrusion, rounded, surface is smooth.Optimum growth temperature is 25~37 DEG C, anaerobism or amphimicrobian,
Optimal pH 6.5 or so is grown muddy in MRS fluid nutrient mediums.
The application also provides a kind of method of freeze-drying lactic acid bacteria, the method includes：
Step 1, by lactic acid bacteria bacterium mud and water, small molecular sugar, protein and L-cysteine salt, sodium alginate and Arab
Glue mixes, stirring, and upper machine liquid is made.
The lactic acid bacteria bacterium mud is the bacterium mud of lactobacillus plantarum strain (Lactobacillus plantarum LP5).Institute
The viable count for stating lactobacillus plantarum strain in bacterium mud (Lactobacillus plantarum LP5) is 1 × 1010CFU/mL with
On.
In a kind of achievable mode, the lactobacillus plantarum strain (Lactobacillus plantarum LP5)
Bacterium mud can be prepared according to method comprising the following steps：
The lactobacillus plantarum (Lactobacillus plantarum LP5) of freezen protective is inoculated in MRS by step 1-1
In fluid nutrient medium, 15h~25h, such as 20h are cultivated at 37 DEG C of temperature.Such secondary culture obtains activated spawn 3 times.
Wherein, MRS fluid nutrient mediums are prepared by following proportioning：10g peptones, 5g beef extracts, 4g yeast extracts, the Portugals 20g
Grape sugar, 2g dipotassium hydrogen phosphates, 5g sodium acetates, 2g trisodium citrates, 1mL Tween 80s, 0.2g magnesium sulfate, 0.05g manganese sulfates are added
1000mL distilled water adjusts pH to 6.5,121 DEG C of sterilizing 15min.
Optionally, the inoculum concentration of the lactobacillus plantarum LP5 is 2% (V/V).
Step 1-2, lactobacillus plantarum LP5 prepared by step 1-1 is inoculated into according to the ratio of 5% (v/v) have by
In the fermentation tank of the MRS fluid nutrient mediums of sterilizing, 37 DEG C of constant temperature incubations, initial pH=6.5 passes through stream plus 25% (V/ of neutralizer
V) ammonium hydroxide control system perseverance pH5.5 after fermentation 3 hours, then stops controlling permanent pH and ferment, when pH is less than 4.5, then passes through
Stream plus 25% (V/V) ammonium hydroxide control system pH to 5.5 fermentation 5h, then spontaneous fermentation is carried out, such loop control 20 hours, until breast
Sour bacterium viable count reaches 1 × 1010CFU/mL or more terminates fermentation.
The spontaneous fermentation refers to 37 DEG C of temperature of control, is fermented under conditions of not control system PH.
The step 1-2 systems prepared are centrifuged step 1-3, extract wet thallus, and bacterium mud is made.
In a kind of achievable mode, the operation temperature of step 1-3 is less than 20 DEG C, so that thalline enters dormant state,
To ensure the activity of thalline.
Further, the rotating speed of centrifuge is 12000~16000rpm, the bacterium under this rotating speed in the centrifugal separation processes
Body cell can supplement and be sufficiently separated out, and somatic cells will not be destroyed.
Under these conditions, the CFU/g of number of viable in the isolated bacterium mud >=300,000,000,000.
The small molecular sugar is selected from sucrose, glucose, lactose, maltose, trehalose, fructose, oligofructose, oligomeric gala
The composition of the weight ratios such as one or more of sugar, preferably sucrose, glucose and lactose.
The inventors discovered that the small molecular sugar, which is added, in system prepared by step 1-2 can reduce in freeze-drying process
Liquid phase forms the size of ice crystal in middle system, to reduce the risk that ice crystal punctures somatic cells wall, and then improves in freeze-dried powder
Number of viable.
The protein is selected from one or more of skimmed milk powder, soyabean protein powder, desalted whey powder, yeast powder,
The preferably composition of skimmed milk powder and yeast powder, wherein the weight ratio of skimmed milk powder and yeast powder is (0.5~2):(0.5~
1.5), such as 0.5:1.
The inventors discovered that the protein can make finished product lactic acid bacteria freeze drying powder form loose character, in order to
The use of lactic acid bacteria freeze drying powder.
One kind in L-cysteine salt, sodium ascorbate, sodium alginate and Arabic gum of the oxygen scavenger or
It is a variety of, preferably L-cysteine salt.In the oxygen scavenger, L-cysteine salt, sodium ascorbate, sodium alginate have also
Oxidizing substance that can be in removing system is added after preparation system in originality, and Arabic gum can then be formed in phage surface
Coated layer so that thalline is isolated with extraneous aerobic environment, to avoid thalline by oxidative damage.
The oxygen scavenger can improve the oxidation resistance of finished product lactic acid bacteria freeze drying powder, improve the steady of lactic acid bacteria freeze drying powder
It is qualitative, extend the shelf life of lactic acid bacteria freeze drying powder.
In step 1, the weight ratio of the bacterium mud, water, small molecular sugar, protein and oxygen scavenger is the weight of small molecular sugar
Amount:The weight of protein:Weight=1 of oxygen scavenger:(5~20):(1~5):(0.1~0.8):(0.5~1.5), preferably 1:
(8~15):(1.5~3.5):(0.3~0.6):(0.8~1.2), such as 1:10:2:0.5:1, wherein the weight of bacterium mud is with bacterium
The total weight of mud.
The upper machine liquid is prepared according to aforementioned proportion, and each component synergistic effect can improve living in lactic acid bacteria freeze drying powder
Bacterium number amount, and can effectively extend the shelf life of lactic acid bacteria freeze drying powder.
After water, small molecular sugar, protein and oxygen scavenger are mixed with lactic acid bacteria bacterium mud, at 15 DEG C~25 DEG C with 50~
The rotating speed of 100rpm stirs 1~5min, and thalline can be made to keep dormant state at 15 DEG C~25 DEG C, mixed with rotating speed stirring
Zoarium system can reduce the shearing force that stirring is formed and be damaged caused by thalline, to number of viable in guarantee system, in stirring 1
System can be stirred evenly after~5min.
Step 2, upper machine liquid prepared by step 1 is made to be down to -45 DEG C~-50 DEG C in 10min, heat preservation.
In a kind of achievable mode, upper machine liquid prepared by step 1 is positioned in lyophilized plate, it is 0.5 to form thickness
The liquid layer of the liquid layer of~2cm, preferably 1~1.5cm thickness.The liquid layer that thickness is 0.5~2cm can be under the action of liquid nitrogen quickly
It is sufficiently cooled to preset temperature, such as -50 DEG C, preset temperature also can be promptly cooled to inside liquid layer, make the liquid in thalline
It is mutually rapidly converted into solid phase, iuntercellular and the intracellular process for forming glassy state are avoided, to intracellular and iuntercellular shape
At the solid phase of rule, and then avoid puncturing cell membrane.
In a kind of achievable mode, the temperature of upper machine liquid is reduced to -50 DEG C in 10min, and thalline is made to keep suspend mode
State.If temperature reduced slowly, -50 DEG C are reduced in 10min or more, then forms that ice crystal is excessive to be easy to puncture cell membrane, such as
The reduction of fruit temperature is too fast, and -50 DEG C are reduced within 5-10min, then is easy to cause material inside and outside non-uniform temperature, causes freezing
Melt during dry, production is caused to fail.
In a kind of achievable mode, the upper machine liquid keeps the temperature 0.2~1h at this temperature after being cooled to -50 DEG C,
Such as 0.5h so that material inside and outside temperature is uniform.If soaking time is more than 1h, energy consumption is wasted；If soaking time is less than
0.2h, then material inside and outside non-uniform temperature.
In a kind of achievable mode, its cooling, liquid nitrogen sheet can be made by way of being passed through liquid nitrogen in upward machine liquid
Body temperature is low, reachable -170 DEG C hereinafter, machine liquid can be made to cool down rapidly, and can starvation, avoid somatic cells
Oxygen injury.
In a kind of achievable mode, the vacuum degree of this step system is 18-22Pa so that material is under cryogenic
Directly distil moisture therein, improves thalline vigor.
Step 3, system prepared by step 2 is placed in -30~-10 DEG C of environment, under vacuum according to temperature programming
It is lyophilized.
In a kind of achievable mode, by system prepared by step 2 by be directly moved in -50 DEG C of environment -30 DEG C~-
In 10 DEG C of environment, as in -20 DEG C of environment, temperature reaches a certain level, and temperature of charge and heating temperature are consistent can be into
The heating of row next stage.
In a kind of achievable mode, the vacuum condition is the condition that vacuum degree is 18Pa~22Pa, in this vacuum
Under degree, ice can directly distil, to achieve the effect that freeze-drying.
In a kind of achievable mode, the system prepared to step 2 according to temperature programming is dried.Optionally, institute
It may include three phases to state temperature programming, is followed successively by 10 DEG C of holdings 25h, 20 DEG C of holdings 20h, 30 DEG C of holding 3h.According to above-mentioned
The mode of temperature programming is dried, and can reduce energy consumption and ensure that obtained finished product lactic acid bacteria freeze drying powder can be dried.
The application also provides a kind of lactobacillus plantarum LP5 bacterium powders prepared by preceding method, the lactobacillus plantarum LP5 bacterium
Viable count of lactobacillus is 4.6 × 10 in powder11CFU/g or more, preservation 360 days, Viable detection are under 25 DEG C of constant temperatures
75.0% or more.Specifically refer to embodiment 1.
Embodiment
Embodiment 1
The commercially available information of agents useful for same is as follows in the present embodiment：
(1) actication of culture and culture
The lactobacillus plantarum (Lactobacillus plantarum LP5, L.plantarum LP5) of freezen protective is connect
Kind in MRS fluid nutrient mediums, 20h is cultivated at 37 DEG C of temperature, such secondary culture obtains activated spawn 3 times；
Wherein, MRS fluid nutrient mediums are prepared by following proportioning：10g peptones, 5g beef extracts, 4g yeast extracts, the Portugals 20g
Grape sugar, 2g dipotassium hydrogen phosphates, 5g sodium acetates, 2g trisodium citrates, 1mL Tween 80s, 0.2g magnesium sulfate, 0.05g manganese sulfates are added
1000mL distilled water adjusts pH to 6.5,121 DEG C of sterilizing 15min.
(2) high density fermentation
Activated lactobacillus plantarum LP5 is inoculated into the MRS cultures having by sterilizing according to the ratio of 5% (v/v)
In the fermentation tank of base, 37 DEG C of constant temperature incubations, initial pH=6.5 passes through stream plus (V/V) ammonium hydroxide of neutralizer 25% control system perseverance
PH5.5 after fermenting 3 hours, then stops controlling permanent pH, continues to ferment, and when pH is less than 4.5, then passes through stream plus 25% (V/
V) the fermentations of ammonium hydroxide control system pH to 5.5 5h, then spontaneous fermentation is carried out, such loop control 20 hours, until viable count of lactobacillus
Reach 1 × 1010CFU/mL or more terminates fermentation.
(3) liquid nitrogen vacuum freeze drying
Reagent described in this step passes through high level electronic beam current and scans material, quickly kills the active bacteria in material
Body, material thickness 2cm, material transmission speed 1-5m/min；
3.1, zymotic fluid made from (2) is centrifuged into extraction wet thallus through 14000rpm, it is 3000 that viable count in bacterium mud, which is made,
~5,000 hundred million CFU/g, operating process feed temperature control are 20 DEG C or less；
3.2,10Kg water, 2Kg sucrose, 0.5Kg skimmed milks and 1.0Kg yeast are added into bacterium mud made from 1Kg steps 3.1
Powder keeps the rotating speed of 100r/min stirrings, under the conditions of 20 DEG C, stirs 2min, and upper machine liquid is made；
3.3, upper machine liquid is put into lyophilized plate, liquid layer thickness 1cm, lyophilized plate is put into liquid nitrogen refrigerating equipment, directly
It is passed through liquid nitrogen and upper machine liquid is cooled to -50 DEG C rapidly in 10min, and keep this temperature 30min；
3.4, it takes out the system after liquid nitrogen quickly cooling and is put into vacuum freeze drier, at this time temperature in vacuum freeze drying cabin
It is -20 DEG C, vacuum freeze drier is by way of radiant temperature, under conditions of keeping vacuum degree 20Pa, is lyophilized 48 hours,
Final products are obtained, partition board heating curve is：10 DEG C, 25h；20 DEG C, 20h；30 DEG C, 3h.
Control group is that the upper machine liquid for preparing step 3.2 is prepared using conventional vacuum freeze-drying method, specially：With
Aforementioned (3) liquid nitrogen vacuum freeze drying is similar, differs only in the operation without 3.3 steps, and body prepared by step 3.2
System directly carries out the processing of 3.4 steps.
Bacterium powder preparation prepared by embodiment 1 and control group is respectively placed under 25 DEG C of constant temperatures, respectively 0 day,
It samples within 30 days, 60 days, 90 days, 120 days, 150 days, 180 days, 210 days, 240 days, 270 days, 300 days, 330 days and 360 days, into
Row viable count is examined, and experimental data is shown in Table 1.
1 liquid nitrogen vacuum freeze drying preparation room temperature bin stability situation of table
As shown in Table 1, the lactic acid bacteria freeze drying pulvis prepared using liquid nitrogen vacuum freeze-drying method provided by the present application is existed
When being stored 360 days under normal temperature condition, viable count can reach 3.51 × 1011CFU/g, survival rate 75.0%, and use general
Under the same conditions, viable count just drops to 2.5 × 10 to freeze dried powder prepared by logical freeze drying process after 30 days11CFU/g with
Under, and viable count is reduced to 1.5 × 10 after 60 days11CFU/g is hereinafter, number of viable has had declined two quantity after 180 days
Grade, this explanation have good bin stability using freeze dried powder prepared by method provided by the present application, have very high reality
Border application value.
The application is described in detail above in association with detailed description and exemplary example, but these explanations are simultaneously
It should not be understood as the limitation to the application.It will be appreciated by those skilled in the art that without departing from the application spirit and scope,
A variety of equivalent substitution, modification or improvements can be carried out to technical scheme and embodiments thereof, these each fall within the application
In the range of.The protection domain of the application is determined by the appended claims.
Claims (10)
1. the lactobacillus plantarum LP5 (Lactobacillus plantarum LP5, L.plantarum LP5) of one plant of separation,
It is characterized in that, microbial preservation number is CGMCC No.13459.
2. a kind of method of freeze-drying lactic acid bacteria, which is characterized in that the method includes：
Step 1, lactic acid bacteria bacterium mud is mixed with water, small molecular sugar, protein and oxygen scavenger, is stirred, upper machine liquid is made；
Step 2, upper machine liquid temperature in 10min prepared by step 1 is made to be down to -45 DEG C~-50 DEG C, heat preservation；
Step 3, system prepared by step 2 is placed in -20 DEG C of environment, is lyophilized under vacuum according to temperature programming.
3. according to the method described in claim 2, it is characterized in that, the lactic acid bacteria bacterium mud is lactobacillus plantarum LP5 bacterium muds.
4. according to the method in claim 2 or 3, which is characterized in that
The small molecular sugar is in sucrose, glucose, lactose, maltose, trehalose, fructose, oligofructose, galactooligosaccharide
One or more, the preferably composition of the weight ratios such as sucrose, glucose and lactose；
The protein is selected from one or more of skimmed milk powder, soyabean protein powder, desalted whey powder, yeast powder, preferably
For the composition of skimmed milk powder and yeast powder；
The oxygen scavenger is selected from one or more of L-cysteine salt, sodium ascorbate, sodium alginate and Arabic gum,
Preferably L-cysteine salt.
5. according to claim 2 to 4 any one of them method, which is characterized in that the bacterium mud, water, small molecular sugar, protein
And the weight that the weight ratio of oxygen scavenger is small molecular sugar:The weight of protein:Weight=1 of oxygen scavenger:(5~20):(1~
5):(0.1~0.8):(0.5~1.5), preferably 1:(8~15):(1.5~3.5):(0.3~0.6):(0.8~1.2), example
Such as 1:10:2:0.5:1.
6. according to claim 2 to 5 any one of them method, which is characterized in that in step 2, by being passed through in upward machine liquid
The mode of liquid nitrogen makes machine liquid cool down.
7. according to claim 2 to 6 any one of them method, which is characterized in that in step 2, the upper machine liquid be cooled to-
After 50 DEG C, 0.2~1h, such as 0.5h are kept the temperature at this temperature.
8. according to claim 2 to 6 any one of them method, which is characterized in that in step 2, the vacuum degree of system is 18-
22Pa。
9. according to claim 2 to 8 any one of them method, which is characterized in that in step 3, the vacuum condition is vacuum
Degree is 18Pa~20Pa；
Described program heating includes three phases, is followed successively by 10 DEG C of holdings 25h, 20 DEG C of holdings 20h, 30 DEG C of holding 3h.
10. a kind of lactic acid bacteria bacterium powder prepared according to any one of claim 2 to 9 the method, which is characterized in that the lactic acid
Bacterium is lactobacillus plantarum LP5, and viable count of lactobacillus is 4.6 × 10 in the bacterium powder11CFU/g or more, under 25 DEG C of constant temperatures
Preservation 360 days, Viable detection are 75.0% or more.
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Cited By (3)
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| CN109497133A (en) * | 2018-11-08 | 2019-03-22 | 江南大学 | A kind of compound multiplication agent for delaying lactobacillus plantarum acidified milk viable count to reduce |
| CN114774281A (en) * | 2022-02-25 | 2022-07-22 | 河南省商业科学研究所有限责任公司 | Composite protective agent for freeze-drying lactobacillus bulgaricus |
| CN114917257A (en) * | 2022-05-19 | 2022-08-19 | 大禺(广州)健康研究有限公司 | Vaginal health probiotic composition, preparation method and application thereof |
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| CN104560802A (en) * | 2014-12-24 | 2015-04-29 | 中国肉类食品综合研究中心 | Fermentation strain and fermentation agent for canned meat products |
| CN106119166A (en) * | 2016-07-11 | 2016-11-16 | 江苏恒顺醋业股份有限公司 | One strain Switzerland lactic acid bacteria and application thereof |
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| EP1145643A1 (en) * | 2000-04-10 | 2001-10-17 | Phillip Connolly | Improved milk protein concentrate |
| CN102597216A (en) * | 2009-10-28 | 2012-07-18 | Cj第一制糖株式会社 | Novel lactobacillus plantarum and composition comprising the same |
| CN104560802A (en) * | 2014-12-24 | 2015-04-29 | 中国肉类食品综合研究中心 | Fermentation strain and fermentation agent for canned meat products |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109497133A (en) * | 2018-11-08 | 2019-03-22 | 江南大学 | A kind of compound multiplication agent for delaying lactobacillus plantarum acidified milk viable count to reduce |
| CN114774281A (en) * | 2022-02-25 | 2022-07-22 | 河南省商业科学研究所有限责任公司 | Composite protective agent for freeze-drying lactobacillus bulgaricus |
| CN114917257A (en) * | 2022-05-19 | 2022-08-19 | 大禺(广州)健康研究有限公司 | Vaginal health probiotic composition, preparation method and application thereof |
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