CN110272837A - A kind of lactobacillus plantarum S3-10 and its screening technique and application - Google Patents
A kind of lactobacillus plantarum S3-10 and its screening technique and application Download PDFInfo
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- CN110272837A CN110272837A CN201811521804.0A CN201811521804A CN110272837A CN 110272837 A CN110272837 A CN 110272837A CN 201811521804 A CN201811521804 A CN 201811521804A CN 110272837 A CN110272837 A CN 110272837A
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- lactobacillus plantarum
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- 240000006024 Lactobacillus plantarum Species 0.000 title claims abstract description 43
- 235000013965 Lactobacillus plantarum Nutrition 0.000 title claims abstract description 43
- 229940072205 lactobacillus plantarum Drugs 0.000 title claims abstract description 43
- 238000000034 method Methods 0.000 title claims abstract description 10
- 238000012216 screening Methods 0.000 title claims abstract description 9
- 235000012055 fruits and vegetables Nutrition 0.000 claims abstract description 9
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims abstract description 8
- 150000003839 salts Chemical class 0.000 claims abstract description 6
- 244000005700 microbiome Species 0.000 claims abstract description 5
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 36
- 241000894006 Bacteria Species 0.000 claims description 23
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 20
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 18
- 239000004310 lactic acid Substances 0.000 claims description 18
- 235000014655 lactic acid Nutrition 0.000 claims description 18
- 239000011780 sodium chloride Substances 0.000 claims description 10
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 9
- 239000007787 solid Substances 0.000 claims description 9
- 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 claims description 8
- 239000008103 glucose Substances 0.000 claims description 8
- 239000002253 acid Substances 0.000 claims description 7
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 6
- 241000186660 Lactobacillus Species 0.000 claims description 6
- 229940039696 lactobacillus Drugs 0.000 claims description 6
- 235000012828 Citrullus lanatus var citroides Nutrition 0.000 claims description 5
- 238000002474 experimental method Methods 0.000 claims description 5
- 108020004465 16S ribosomal RNA Proteins 0.000 claims description 4
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 claims description 4
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 4
- 229930006000 Sucrose Natural products 0.000 claims description 4
- 235000021016 apples Nutrition 0.000 claims description 4
- 239000011575 calcium Substances 0.000 claims description 4
- 229910052791 calcium Inorganic materials 0.000 claims description 4
- 235000013336 milk Nutrition 0.000 claims description 4
- 239000008267 milk Substances 0.000 claims description 4
- 210000004080 milk Anatomy 0.000 claims description 4
- 238000004321 preservation Methods 0.000 claims description 4
- 239000005720 sucrose Substances 0.000 claims description 4
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 3
- XJRPTMORGOIMMI-UHFFFAOYSA-N ethyl 2-amino-4-(trifluoromethyl)-1,3-thiazole-5-carboxylate Chemical compound CCOC(=O)C=1SC(N)=NC=1C(F)(F)F XJRPTMORGOIMMI-UHFFFAOYSA-N 0.000 claims description 3
- 238000010186 staining Methods 0.000 claims description 3
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 2
- 241000726221 Gemma Species 0.000 claims description 2
- 102000003992 Peroxidases Human genes 0.000 claims description 2
- 239000001110 calcium chloride Substances 0.000 claims description 2
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 2
- 238000007865 diluting Methods 0.000 claims description 2
- 239000002054 inoculum Substances 0.000 claims description 2
- 238000009630 liquid culture Methods 0.000 claims description 2
- 108040007629 peroxidase activity proteins Proteins 0.000 claims description 2
- 244000141359 Malus pumila Species 0.000 claims 2
- 244000241235 Citrullus lanatus Species 0.000 claims 1
- 150000002475 indoles Chemical class 0.000 claims 1
- 230000001580 bacterial effect Effects 0.000 abstract description 14
- 235000013406 prebiotics Nutrition 0.000 abstract 1
- 230000004083 survival effect Effects 0.000 description 15
- 239000002609 medium Substances 0.000 description 11
- 235000013361 beverage Nutrition 0.000 description 6
- 238000000855 fermentation Methods 0.000 description 6
- 230000004151 fermentation Effects 0.000 description 6
- 241000220225 Malus Species 0.000 description 5
- 241000219109 Citrullus Species 0.000 description 4
- 235000013399 edible fruits Nutrition 0.000 description 4
- 239000012530 fluid Substances 0.000 description 4
- 235000015097 nutrients Nutrition 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 235000013311 vegetables Nutrition 0.000 description 4
- 238000005138 cryopreservation Methods 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 235000016709 nutrition Nutrition 0.000 description 3
- 230000035764 nutrition Effects 0.000 description 3
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- SIKJAQJRHWYJAI-UHFFFAOYSA-N Indole Chemical compound C1=CC=C2NC=CC2=C1 SIKJAQJRHWYJAI-UHFFFAOYSA-N 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000005418 vegetable material Substances 0.000 description 2
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- RWSOTUBLDIXVET-UHFFFAOYSA-N Dihydrogen sulfide Chemical compound S RWSOTUBLDIXVET-UHFFFAOYSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 101100107522 Mus musculus Slc1a5 gene Proteins 0.000 description 1
- 230000003796 beauty Effects 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 235000013365 dairy product Nutrition 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 235000013402 health food Nutrition 0.000 description 1
- 229910000037 hydrogen sulfide Inorganic materials 0.000 description 1
- PZOUSPYUWWUPPK-UHFFFAOYSA-N indole Natural products CC1=CC=CC2=C1C=CN2 PZOUSPYUWWUPPK-UHFFFAOYSA-N 0.000 description 1
- RKJUIXBNRJVNHR-UHFFFAOYSA-N indolenine Natural products C1=CC=C2CC=NC2=C1 RKJUIXBNRJVNHR-UHFFFAOYSA-N 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 235000013622 meat product Nutrition 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 239000008239 natural water Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000002864 sequence alignment Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
- A23L2/382—Other non-alcoholic beverages fermented
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/52—Adding ingredients
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/52—Adding ingredients
- A23L2/60—Sweeteners
-
- 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
- C12N1/02—Separating microorganisms from their culture media
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- 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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- 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
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/25—Lactobacillus plantarum
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Abstract
The invention discloses a kind of lactobacillus plantarum S3-10 and its screening technique and application, the lactobacillus plantarum S3-10 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and deposit number is CGMCC No.16751.The bacterial strain has good acidproof, salt tolerant, resistance to sugared characteristic, has broad prospect of application in terms of fermented fruits and vegetables juice, the prebiotic product of exploitation.
Description
Technical field
The present invention relates to food microorganisms technical fields, and in particular to a kind of lactobacillus plantarum S3-10 and its screening technique
And application.
Background technique
Lactic acid bacteria refers to that energy fermentable carbohydrates (mainly glucose) produce one kind of a large amount of lactic acid in nature
In widely distributed gram-positive bacterium common name.Due to lactic acid bacteria have nutrition, health special efficacy, it is fashionable Europe,
The markets such as beauty, Japan and Korea S., and it is widely used in the food such as health food, beverage, meat products, dairy products and preventive medicine field.
Due to the increasingly raising of the living standard, the level of consumption of the development and people of society, domestic beverage market occurs
Earth-shaking variation, people gradually pursue the beverage of natural health and nutrition.Some countries such as moral, Japan and Korea S. are already to hair
Ferment beverage made of fruits or vegetables has conducted extensive research and develops, especially Japanese, and the development trend in beverage made of fruits or vegetables market comes into
Good track.Domestic researcher lactobacillus-fermented also has been carried out with regard to some fruit and vegetable materials in terms of exploration, and at the beginning of achieving
The progress of step.Lactic acid bacteria in research for fruits and vegetables fermentation is mostly milk fermentation bacterial strain, limited and acidproof using fruits and vegetables ability
Property it is very poor, therefore the fruit and vegetable materials that can not effectively ferment generate good flavor and nutriment.Therefore, screening is suitable for fruit
The tailored version lactic acid bacteria of vegetable fermentation can shorten fruits and vegetables fermentation time, produce more nutrition, taste is better, the higher fruit of safety
Vegetable fermented product.
Summary of the invention
The present invention is in view of the above-mentioned problems, provide a kind of lactobacillus plantarum S3-10 and its screening technique and using
Goal of the invention is realized by following scheme: a kind of lactobacillus plantarum S3-10(Lactobacillus plantarum S3-
10) China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number, are preserved on November 16th, 2018
For CGMCC NO. 16751.
Further, lactobacillus plantarum S3-10(Lactobacillus plantarumS3-10 morphological feature) are as follows:
Circular colonies, protrusion, smooth, fine and closely woven, faint yellow or canescence, Gram-positive, no gemma, width be 0.9 ~ 1.2 micron,
A length of 3 ~ 8 microns of rod-short, individually, in pairs or at short chain.
Further, lactobacillus plantarum S3-10(Lactobacillus plantarumS3-10 16S rDNA total order)
Column are as shown in SEQ ID NO.1.
Further, lactobacillus plantarum S3-10(Lactobacillus plantarumS3-10) have stronger resistance to
The characteristic of acid, salt tolerant, resistance to sugar.
Further, lactobacillus plantarum S3-10(Lactobacillus plantarumS3-10) in fermented fruits and vegetables juice
Application.Zymotechnique formula are as follows: watermelon juice 33.3%, glucose 5%, sucrose 5%, calcium chloride 0.5%, sodium chloride 3%, citric acid
0.1%.Lactobacillus plantarum S3-10 is inoculated into above-mentioned formula with 5% inoculum concentration, after 37 DEG C of 18 h of constant-temperatureanaerobic anaerobic culture, produces acid
Amount is 18.63 mol/L, and viable count is maintained at (2.00-2.20) × 109 cfu/mL。
A kind of method of separating plant lactobacillus, takes the apple sample in Yiyuan Apples In Mountainous Area producing region, crushes and inhales after diluting
It takes in solution coating to the MRS solid medium containing bromocresol purple, 37 DEG C of 24 h of constant-temperatureanaerobic anaerobic culture；It selects raised, yellowish
The culture in acid MRS solid medium (pH 2.0) of color bacterium colony carries out primary election, then selects target bacterium colony and continues in MRS with high salt
Final election is carried out in solid medium (addition 10%NaCl and 0.5% calcium carbonate), the apparent single colonie of yellow calcium circle is selected and continuously passes
It is commissioned to train and supports no less than 3 times, the single colonie purified.It selects the single bacterium and carries out Gram's staining, peroxidase haptoreaction, Yin
Diindyl experiment, litmus milk experiment and lactic acid chromatography etc. carry out the identification of lactic acid bacteria.Sterile glycerol is added in lactic acid bacteria after identification
Liquid Culture is based on -80 DEG C and saves backup.
Detailed description of the invention
Fig. 1 is the colony characteristics figure of the bacterial strain of the isolated lactobacillus plantarum S3-10 of the present invention, and Fig. 2, Fig. 3 are
The micro- sem observation characteristic pattern of Gram's staining of the bacterial strain of the isolated lactobacillus plantarum S3-10 of the present invention.
Specific embodiment
Below with reference to embodiment, the present invention is further explained, but not limited to the examples.Reality as used in the following examples
Proved recipe method is conventional method unless otherwise specified.The materials, reagents and the like used in the following examples, unless otherwise specified,
Commercially obtain.
Embodiment 1: this example demonstrates that the separation and identification of lactobacillus plantarum S3-10
One, the separation of bacterial strain
Present invention bacterium from the microbiologic population in Yiyuan Apples In Mountainous Area producing region.
The apple sample in Yiyuan Apples In Mountainous Area producing region is taken, crushes and gradient dilution is 10-8Afterwards draw solution be applied to containing
In the MRS solid medium of bromocresol purple, 37 DEG C of 24 h of constant-temperatureanaerobic anaerobic culture；It is solid in acid MRS to select protrusion, faint yellow bacterium colony
Culture carries out primary election in body culture medium (pH 2.0), then selects target bacterium colony and continues in MRS solid medium with high salt (addition
10%NaCl and 0.5% calcium carbonate) in carry out final election, select the apparent single colonie continuous passage culture of yellow calcium circle and be no less than 3 times,
The single colonie purified.Picking individual colonies are inoculated in 37 DEG C of 18 h of culture in MRS fluid nutrient medium in case preservation and identification make
With.
Two, the identification of bacterial strain
Pure cultured spawn Gram-positive, microscopy be it is rod-shaped, hydrogen peroxide test, indole reaction, gelatin liquefaction experiment and
Hydrogen sulfide reaction result is feminine gender.Litmus milk tests and lactic acid chromatography test for identification are the result shows that the bacterial strain energy lactic acid producing is
Lactic acid bacteria.The aimed strain of purifying is subjected to the identification of 16S rDNA gene sequencing, as a result as shown in SEQ ID No.1, in NCBI
BLAST sequence alignment is carried out on website, as the result is shown the sequence and lactobacillus plantarum (Lactobacillus plantarum)
16S rDNA sequence homology is more than 99%.
In summary as a result, determine screening S3-10 bacterial strain be lactobacillus plantarum (Lactobacillus plantarum).The Strain Designation is lactobacillus plantarum S3-10(by applicantLactobacillus plantarum S3-
10) China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation, and are preserved on November 16th, 2018
Number be CGMCC No. 16751.
Embodiment 2: this example demonstrates that the resistance to sugar test of the acidproof salt tolerant of lactobacillus plantarum S3-10
Lactobacillus plantarum S3-10 glycerol cryopreservation tube is activated to cell concentration and is adjusted to 2.5 × 108, being then inoculated with pH is 2.0-
3.0 MRS fluid nutrient medium, 37 DEG C of constant temperature incubations are pair with 0 h sample in the separately sampled carry out count plate of 0 h and 3 h
According to the survival rate of bacterial strain, survival rate=(viable count of 3 h/0 h viable count) × 100% in calculating sample
| Bacterial strain | Survival rate in the MRS of pH=2.0 | Survival rate in the MRS of pH=2.5 | Survival rate in the MRS of pH=3.0 |
| Lactobacillus plantarum S3-10 | 90.1% | 107.5% | 129.1% |
| Compare lactic acid bacteria | 54.2% | 88.7% | 94.5% |
Lactobacillus plantarum S3-10 glycerol cryopreservation tube is activated to cell concentration and is adjusted to 2.5 × 108, is then inoculated with NaCl concentration
For the MRS fluid nutrient medium of 5.0%-10.0%, 37 DEG C of constant temperature incubations, in the separately sampled carry out count plate of 0 h and 3 h, with 0 h
Sample is control, calculates the survival rate of bacterial strain in sample, survival rate=(viable count of 3 h/0 h viable count) × 100%
| Bacterial strain | Survival rate in the MRS containing 5.0%NaCl | Survival rate in the MRS containing 8.0%NaCl | Survival rate in the MRS containing 10.0%NaCl |
| Lactobacillus plantarum S3-10 | 108.7% | 96.7% | 89.6% |
| Compare lactic acid bacteria | 95.8% | 80.4% | 62.3% |
Lactobacillus plantarum S3-10 glycerol cryopreservation tube is activated to cell concentration and is adjusted to 2.5 × 108, it is dense to be then inoculated with glucose
Degree is the MRS fluid nutrient medium of 5.0%-10.0%, 37 DEG C of constant temperature incubations, in the separately sampled carry out count plate of 0 h and 3 h, with 0
H sample is control, calculates the survival rate of bacterial strain in sample, survival rate=(viable count of 3 h/0 h viable count) × 100%
| Bacterial strain | Survival rate in the MRS containing 5.0% glucose | Survival rate in the MRS containing 8.0% glucose | Survival rate in the MRS containing 10.0% glucose |
| Lactobacillus plantarum S3-10 | 108.7% | 96.7% | 89.6% |
| Compare lactic acid bacteria | 97.6% | 88.5% | 70.1% |
Embodiment 3: this example demonstrates that lactobacillus plantarum S3-10 fermented fruits and vegetables juice
The preparation of lactobacillus plantarum S3-10 seed liquor: the lactobacillus plantarum S3-10 glycerol stocks pipe of preservation is inoculated into MRS liquid
In body culture medium, the seed liquor of lactobacillus plantarum S3-10 is obtained after 37 DEG C of 18 h of constant-temperatureanaerobic anaerobic culture.
It configures liquid fermentation medium: watermelon juice filtering is selected, according to watermelon juice 33.3%, glucose 5%, sucrose 5%, chlorine
Change calcium 0.5%, sodium chloride 3%, citric acid 0.1% proportion configuration, 121 DEG C, sterilizing 30 min after, be cooled to 37 DEG C or so.
The seed liquor of 5% lactobacillus plantarum S3-10 is inoculated into liquid fermentation medium, 37 DEG C of constant-temperatureanaerobic anaerobic cultures
After 18 h, producing acid amount is 18.63 mol/L, and viable count is maintained at (2.00-2.20) × 109cfu/mL.4 DEG C of refrigerators are put into protect
6 h of temperature, carry out taste allotment with sodium citrate and sucrose, and the strong lactic acid bacteria hair of tasty, natural water melon taste can be obtained
Ferment beverage.
Sequence table
<110>Shandong Technology Univ
<120>a kind of lactobacillus plantarum S3-10 and its screening technique and application
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1474
<212> DNA
<213>lactobacillus plantarum S3-10 (Lactobacillus plantarum S3-10)
<400> 1
gggccgtggc ggcgtgctat acatgcaagt cgaacgaact ctggtattga ttggtgcttg 60
catcatgatt tacatttgag tgagtggcga actggtgagt aacacgtggg aaacctgccc 120
agaagcgggg gataacacct ggaaacagat gctaataccg cataacaact tggaccgcat 180
ggtccgagct tgaaagatgg cttcggctat cacttttgga tggtcccgcg gcgtattagc 240
tagatggtgg ggtaacggct caccatggca atgatacgta gccgacctga gagggtaatc 300
ggccacattg ggactgagac acggcccaaa ctcctacggg aggcagcagt agggaatctt 360
ccacaatgga cgaaagtctg atggagcaac gccgcgtgag tgaagaaggg tttcggctcg 420
taaaactctg ttgttaaaga agaacatatc tgagagtaac tgttcaggta ttgacggtat 480
ttaaccagaa agccacggct aactacgtgc cagcagccgc ggtaatacgt aggtggcaag 540
cgttgtccgg atttattggg cgtaaagcga gcgcaggcgg ttttttaagt ctgatgtgaa 600
agccttcggc tcaaccgaag aagtgcatcg gaaactggga aacttgagtg cagaagagga 660
cagtggaact ccatgtgtag cggtgaaatg cgtagatata tggaagaaca ccagtggcga 720
aggcggctgt ctggtctgta actgacgctg aggctcgaaa gtatgggtag caaacaggat 780
tagataccct ggtagtccat accgtaaacg atgaatgcta agtgttggag ggtttccgcc 840
cttcagtgct gcagctaacg cattaagcat tccgcctggg gagtacggcc gcaaggctga 900
aactcaaagg aattgacggg ggcccgcaca agcggtggag catgtggttt aattcgaagc 960
tacgcgaaga accttaccag gtcttgacat actatgcaaa tctaagagat tagacgttcc 1020
cttcggggac atggatacag gtggtgcatg gttgtcgtca gctcgtgtcg tgagatgttg 1080
ggttaagtcc cgcaacgagc gcaaccctta ttatcagttg ccagcattaa gttgggcact 1140
ctggtgagac tgccggtgac aaaccggagg aaggtgggga tgacgtcaaa tcatcatgcc 1200
ccttatgacc tgggctacac acgtgctaca atggatggta caacgagttg cgaactcgcg 1260
agagtaagct aatctcttaa agccattctc agttcggatt gtaggctgca actcgcctac 1320
atgaagtcgg aatcgctagt aatcgcggat cagcatgccg cggtgaatac gttcccgggc 1380
cttgtacaca ccgcccgtca caccatgaga gtttgtaaca cccaaagtcg gtggggtaac 1440
cttttaggaa ccagccgcta aaggggaaca aaat 1474
Claims (8)
1. a kind of lactobacillus plantarum S3-10(Lactobacillus plantarumS3-10) it is preserved in Chinese microorganism strain
Preservation administration committee common micro-organisms center, deposit number are CGMCC No.16751.
2. lactobacillus plantarum S3-10 according to claim 1, it is characterised in that: circular colonies, protrusion is smooth, fine and closely woven,
Faint yellow or canescence, Gram-positive, no gemma, width are 0.9 ~ 1.2 micron, a length of 3 ~ 8 microns of rod-short, list
It is a, in pairs or at short chain.
3. lactobacillus plantarum S3-10 according to claim 1, it is characterised in that: the lactobacillus plantarum 16S rDNA
As shown in SEQ ID NO.1.
4. a kind of screening technique for separating lactobacillus plantarum described in claim 1, it is characterised in that: Yiyuan Apples In Mountainous Area is taken to produce
The apple sample in area, draw solution is applied in the MRS solid medium containing bromocresol purple after crushing and diluting, 37 DEG C of constant temperature
24 h of Anaerobic culturel；It selects the culture in acid MRS solid medium (pH 2.0) of protrusion, faint yellow bacterium colony and carries out primary election, so
After select target bacterium colony and continue to carry out final election in MRS solid medium with high salt (addition 10%NaCl and 0.5% calcium carbonate), select
The apparent single colonie continuous passage culture of yellow calcium circle is no less than 3 times, the single colonie purified.
5. selecting the single bacterium carries out Gram's staining, peroxidase haptoreaction, indoles experiment, litmus milk experiment and lactic acid layer
Analysis etc. carries out the identification of lactic acid bacteria.
It is saved backup 6. sterile glycerol Liquid Culture is added in the lactic acid bacteria after identification and is based on -80 DEG C.
7. a kind of application of lactobacillus plantarum S3-10 according to claim 1 in fermented fruits and vegetables juice, it is characterised in that institute
The zymotechnique formula stated are as follows: watermelon juice 33.3%, glucose 5%, sucrose 5%, calcium chloride 0.1%, sodium chloride 3%, citric acid
0.1%。
8. lactobacillus plantarum S3-10 is inoculated into above-mentioned formula with 5% inoculum concentration, after 37 DEG C of 18 h of constant-temperatureanaerobic anaerobic culture, produce
Acid amount is 18.63 mol/L, and viable count is maintained at (2.00-2.20) × 109 cfu/mL。
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| CN112553124A (en) * | 2020-12-28 | 2021-03-26 | 福建省农业科学院农业工程技术研究所 | Lactobacillus plantarum strain and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107058159A (en) * | 2016-12-29 | 2017-08-18 | 石家庄君乐宝乳业有限公司 | There are the Lactobacillus plantarum JMCC0017, its screening technique and application of good survival in milk beverage |
| CN107227278A (en) * | 2017-07-17 | 2017-10-03 | 浙江鸣食品股份有限公司 | A kind of Lactobacillus plantarum A11 and its application |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107058159A (en) * | 2016-12-29 | 2017-08-18 | 石家庄君乐宝乳业有限公司 | There are the Lactobacillus plantarum JMCC0017, its screening technique and application of good survival in milk beverage |
| CN107227278A (en) * | 2017-07-17 | 2017-10-03 | 浙江鸣食品股份有限公司 | A kind of Lactobacillus plantarum A11 and its application |
Non-Patent Citations (1)
| Title |
|---|
| 张玉慧: "乳酸菌发酵蓝莓果汁的工艺研究", 《中国优秀博硕士学位论文全文数据库(硕士) 工程科技Ⅰ辑》 * |
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| CN112553124A (en) * | 2020-12-28 | 2021-03-26 | 福建省农业科学院农业工程技术研究所 | Lactobacillus plantarum strain and application thereof |
| CN112553124B (en) * | 2020-12-28 | 2022-06-07 | 福建省农业科学院农业工程技术研究所 | Lactobacillus plantarum strain and application thereof |
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