CN108315279A - A kind of long-time storage method of lactobacillus plantarum liquid preparation - Google Patents
A kind of long-time storage method of lactobacillus plantarum liquid preparation Download PDFInfo
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- CN108315279A CN108315279A CN201810309582.XA CN201810309582A CN108315279A CN 108315279 A CN108315279 A CN 108315279A CN 201810309582 A CN201810309582 A CN 201810309582A CN 108315279 A CN108315279 A CN 108315279A
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- 239000007788 liquid Substances 0.000 title claims abstract description 34
- 240000006024 Lactobacillus plantarum Species 0.000 title claims abstract description 22
- 235000013965 Lactobacillus plantarum Nutrition 0.000 title claims abstract description 22
- 229940072205 lactobacillus plantarum Drugs 0.000 title claims abstract description 22
- 238000000034 method Methods 0.000 title claims abstract description 21
- 238000003860 storage Methods 0.000 title claims abstract description 18
- 238000002360 preparation method Methods 0.000 title claims abstract description 17
- 241000894006 Bacteria Species 0.000 claims abstract description 31
- 241000186660 Lactobacillus Species 0.000 claims abstract description 27
- 229940039696 lactobacillus Drugs 0.000 claims abstract description 25
- 239000007787 solid Substances 0.000 claims abstract description 9
- 239000000463 material Substances 0.000 claims abstract description 8
- 239000000654 additive Substances 0.000 claims description 6
- 230000000996 additive effect Effects 0.000 claims description 6
- 239000000725 suspension Substances 0.000 abstract description 5
- 239000004615 ingredient Substances 0.000 abstract description 4
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 20
- 238000012360 testing method Methods 0.000 description 16
- 239000004310 lactic acid Substances 0.000 description 10
- 235000014655 lactic acid Nutrition 0.000 description 10
- 238000004321 preservation Methods 0.000 description 5
- 239000003223 protective agent Substances 0.000 description 4
- 229920002472 Starch Polymers 0.000 description 3
- 230000001580 bacterial effect Effects 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000000855 fermentation Methods 0.000 description 3
- 230000004151 fermentation Effects 0.000 description 3
- 239000001814 pectin Substances 0.000 description 3
- 235000010987 pectin Nutrition 0.000 description 3
- 229920001277 pectin Polymers 0.000 description 3
- 235000010408 potassium alginate Nutrition 0.000 description 3
- 239000000737 potassium alginate Substances 0.000 description 3
- MZYRDLHIWXQJCQ-YZOKENDUSA-L potassium alginate Chemical compound [K+].[K+].O1[C@@H](C([O-])=O)[C@@H](OC)[C@H](O)[C@H](O)[C@@H]1O[C@@H]1[C@@H](C([O-])=O)O[C@@H](O)[C@@H](O)[C@H]1O MZYRDLHIWXQJCQ-YZOKENDUSA-L 0.000 description 3
- 239000008107 starch Substances 0.000 description 3
- 235000019698 starch Nutrition 0.000 description 3
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 241000219071 Malvaceae Species 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 238000011031 large-scale manufacturing process Methods 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 241000208479 Anagallis arvensis Species 0.000 description 1
- 235000001405 Artemisia annua Nutrition 0.000 description 1
- 240000000011 Artemisia annua Species 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 239000003674 animal food additive Substances 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000000151 deposition Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000013401 experimental design Methods 0.000 description 1
- 235000021121 fermented vegetables Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 244000005709 gut microbiome Species 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 241000411851 herbal medicine Species 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 229940099596 manganese sulfate Drugs 0.000 description 1
- 235000007079 manganese sulphate Nutrition 0.000 description 1
- 239000011702 manganese sulphate Substances 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 235000006286 nutrient intake Nutrition 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000005057 refrigeration Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 238000004114 suspension culture Methods 0.000 description 1
- 235000019640 taste Nutrition 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 description 1
- 239000001393 triammonium citrate Substances 0.000 description 1
- 235000011046 triammonium citrate Nutrition 0.000 description 1
- 238000009777 vacuum freeze-drying Methods 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/04—Preserving or maintaining viable microorganisms
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- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
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Abstract
The invention discloses a kind of long-time storage methods of lactobacillus plantarum liquid preparation, include the following steps：(1) the purified culture of lactobacillus plantarum is handled, lactobacillus liquid bacterium solution is made；(2) solid MRS formula materials are added into lactobacillus liquid bacterium solution；(3) the liquid lactobacillus for adding solid MRS formula materials is stored in low temperature or under room temperature.The present invention adds solid MRS ingredients in lactobacillus bacterium solution and can effectively extend the holding time of lactobacillus suspension body preparation in conjunction with suitable temperature condition (low temperature or room temperature), it is ensured that certain viable count of lactobacillus suspension body preparation；Storage method advantages of simple of the present invention, it is swift to operate, it has a good application prospect.
Description
Technical field
The present invention relates to a kind of long-time storage methods of lactobacillus plantarum liquid preparation.
Background technology
Lactobacillus plantarum is one kind of lactic acid bacteria, and optimum growth temperature is 30 DEG C~35 DEG C, and anaerobism or amphimicrobian are most suitable
PH 6.5 or so, belongs to homofermentative lactic bacteria.The life of lactobacillus plantarum and the mankind are in close relations, are that one kind being common in milk
Lactic acid bacteria in oil, meat and many fermented vegetable products can be colonized in enteron aisle and play beneficial effect.
Lactobacillus plantarum has a major impact intestinal microflora, in food fermentation, industrial lactic fermentation and medical treatment
The fields such as health care all have very strong application potential.In recent years, as food, feed etc. need the product of large-scale production to fermentation
The requirement of technique steps up, and the efficient high vigor for groping fermenting additive preserves and safe handling just seems more important.So
And lactic acid bacteria vigor is weaker, preserves more difficulty, so far, most effective store method is still vacuum freeze-drying method,
This method can be such that lactic acid bacteria preserves 3 months or more.But the method is only applicable to the preservation of strain and difficulty is used for production practices.With
The increase year by year of lactobacillus feed additive usage amount is badly in need of solving the problems, such as that the high-volume of lactic acid bacteria, room temperature, high vigor preserve,
To lay the foundation for large-scale production as feed addictive for microorganism.For this purpose, researchers have carried out multi-party research,
If Wang Chengtao et al. has studied influence of the five tastes Chinese herbal medicine to lactobacter growth and preservation vigor, discovery preserves l under room temperature
The culture solution of 1% sweet wormwood is added in week, 2 weeks, 4 weeks lactobacillus plantarum LP-2, and 2.5 are increased than control group viable count after preserving 4 weeks
Times；Lin Yanwen has studied influence of the common bombax flower to lactobacter growth and preservation vigor, finds after preserving 3 weeks, contains 5% common bombax flower
Viable count in the lactobacillus plantarum liquid of concentration is 16 times of blank control.However, these research do not provide preservation after with guarantor
The comparable situation of viable count before depositing, and the duration for studying observation fall short of, practical operation there is also certain difficulty, because
This has no practical application.
Invention content
In view of this, the purpose of the present invention is to provide a kind of long-time storage methods of lactobacillus plantarum liquid preparation.
In order to achieve the above objectives, the present invention provides the following technical solutions：
A kind of long-time storage method of lactobacillus plantarum liquid preparation, includes the following steps：
(1) the purified culture of lactobacillus plantarum is handled, lactobacillus liquid bacterium solution is made；
(2) solid MRS formula materials are added into lactobacillus liquid bacterium solution；
(3) the liquid lactobacillus for adding solid MRS formula materials is stored in low temperature or under room temperature.
Preferably, in step (1) the lactobacillus liquid bacterium solution lactobacillus a concentration of 95 × 108CFU/mL~121 ×
108CFU/mL。
Preferably, the additive amount of the MRS and the mass volume ratio of lactobacillus liquid bacterium solution are 2~3%.
Preferably, the additive amount of the MRS and the mass volume ratio of lactobacillus liquid bacterium solution are 3%.
Preferably, the cryogenic conditions are 4 DEG C, and the room temperature condition is 20 ± 5 DEG C.
The beneficial effects of the present invention are：
The present invention adds solid MRS ingredients in lactobacillus bacterium solution and combines suitable temperature condition (low temperature or room temperature) can
Effectively extend the holding time of lactobacillus suspension body preparation, it is ensured that certain viable count of lactobacillus suspension body preparation；Storage of the present invention
Method advantages of simple is deposited, it is swift to operate, it has a good application prospect.
Specific implementation mode
Below by specific embodiment, the invention will be further described, so that those skilled in the art can be better
Understand the present invention and can be practiced, but illustrated embodiment is not as a limitation of the invention.
Embodiment 1
Test material needed for 1.1 present invention：Lactobacillus plantarum, MRS culture mediums (every liter of ingredient：Peptone 10.0g, beef
Powder 5.0, yeast 4.0g, glucose 20.0g, Tween 80 1.0mL, dipotassium hydrogen phosphate 2.0g, sodium acetate 5.0g, Triammonium citrate
2.0g, magnesium sulfate 0.2g, manganese sulfate 0.05g), potassium alginate, starch, pectin.
1.2 test method：
Experiment is divided into 8 groups, and test group 1 and test group 5 are that liquid bacterium solution, test group 2 and test group 6 are added for liquid bacterium solution
3%MRS formula materials (m/v, mass volume ratio), test group 3 and test group 7 be liquid bacterium solution add 1% potassium alginate (m/v,
Mass volume ratio), test group 4 and test group 8 are that liquid bacterium solution adds the starch of 2% (m/v, mass volume ratio) and pectin mixes
Object, between the two 1:1, it is both needed to sterilize before substance addition added by every group.
After lactobacillus suspension culture good (every grade culture 14h or so), viable plate count, then according to different designs at
Reason, each processing are sub-packed in 12 centrifuge tubes (1.5mL), and sealed membrane sealing deposits in 4 degree of refrigerators and room temperature, respectively
Count plate was carried out at the 7th, 14,21,28,35,42 day.
1.3 experimental designs and experimental result is shown in Tables 1 and 2s
Table 1
Table 2
1.4 interpretation of result
(1) lactobacillus plantarum strain quality used in this experiment belongs to medium, and miscellaneous bacteria is more when diluting bacterium powder coated plate culture, warp
After crossing purifying culture, 10,000,000,000 or so (test group detects on the 1st day) can be grown in MRS fluid nutrient mediums.
(2) protective agent potassium alginate, starch, pectin used in testing are the classical protective agent of lactic acid bacteria freeze drying process, this
The results show, during fluid storage, the effect is unsatisfactory for these protective agents.
(3) solid MRS ingredients are added in lactic acid bacterial liquid can extend shelf life the preservation situation of a bacterium number, wherein 4 DEG C
When storage, lactic acid bacteria is dead with bacterial apolexis after adding MRS, and novel bacterial is slowly proliferated, and when arriving off-test, there is 1-2 hundred million
Viable count (test group 2).When normal temperature storage, since proper temperature lactic acid bacteria bacterium number is proliferated rapidly, (test group 6 is examined on the 7th day
Survey), but the later stage decline rapidly death with nutrient consumption strain.The additive amount of MRS is advisable with 2-3% (m/v), can excessively be caused
Bacterium solution is sticky.
(4) consider from deisgn product angle, if being weighed with the shelf-life, it is to guarantee the quality that refrigeration+MRS modes, which may be selected, with 35 days
Phase, the about viable bacteria of 5-6 hundred million (test group 2 detects on the 35th day).If with cost and convenience measurement can select room temperature+
MRS, using two weeks as the shelf-life, about 1,000,000,000 grades of viable count (test group 6 detects on the 14th day).
To sum up, embodiment 1 is the results show that storage method provided by the invention can be carried significantly relative to the classical protective agent of freeze-drying
High liquid Lactobacillus Survival.Meanwhile experiment also turns out, is stored under lower temperature conditions, lactic acid bacteria keeps athermobiosis shape
State, it is advantageously ensured that certain viable count of lactobacillus suspension body preparation.
It should be noted that in the case that room temperature of the present invention or room temperature are not particularly illustrated, generally referred to as 20
±5℃。
Embodiment described above is only to absolutely prove preferred embodiment that is of the invention and being lifted, protection model of the invention
It encloses without being limited thereto.Those skilled in the art on the basis of the present invention made by equivalent substitute or transformation, in the present invention
Protection domain within.Protection scope of the present invention is subject to claims.
Claims (6)
1. a kind of long-time storage method of lactobacillus plantarum liquid preparation, which is characterized in that include the following steps：
(1) the purified culture of lactobacillus plantarum is handled, lactobacillus liquid bacterium solution is made；
(2) solid MRS formula materials are added into lactobacillus liquid bacterium solution；
(3) the liquid lactobacillus for adding solid MRS formula materials is stored in low temperature or under room temperature.
2. the long-time storage method of lactobacillus plantarum liquid preparation according to claim 1, which is characterized in that the step
Suddenly in (1) lactobacillus liquid bacterium solution lactobacillus a concentration of 95 × 108CFU/mL~121 × 108CFU/mL。
3. the long-time storage method of lactobacillus plantarum liquid preparation according to claim 1, which is characterized in that described
The additive amount of MRS is 2~3% with the mass volume ratio of lactobacillus liquid bacterium solution.
4. the long-time storage method of lactobacillus plantarum liquid preparation according to claim 1, which is characterized in that described
The additive amount of MRS is 3% with the mass volume ratio of lactobacillus liquid bacterium solution.
5. the long-time storage method of lactobacillus plantarum liquid preparation according to claim 1, which is characterized in that described low
Warm condition is 4 DEG C.
6. the long-time storage method of lactobacillus plantarum liquid preparation according to claim 1, which is characterized in that the room
Warm condition is 20 ± 5 DEG C.
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| US20050208630A1 (en) * | 2002-06-03 | 2005-09-22 | Luc De Vuyst | Novel streptococcus thermophilus strains producing stable high-molecular-mass exopolysaccharides |
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| CN106520633A (en) * | 2016-12-15 | 2017-03-22 | 汉臣氏（沈阳）儿童制品有限公司 | Method for preparing lactobacillus plantarum freeze-dried powder |
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2018
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Application publication date: 20180724 |