CN107227278A - A kind of Lactobacillus plantarum A11 and its application - Google Patents
A kind of Lactobacillus plantarum A11 and its application Download PDFInfo
- Publication number
- CN107227278A CN107227278A CN201710580930.2A CN201710580930A CN107227278A CN 107227278 A CN107227278 A CN 107227278A CN 201710580930 A CN201710580930 A CN 201710580930A CN 107227278 A CN107227278 A CN 107227278A
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- Prior art keywords
- lactobacillus plantarum
- acid
- application
- fermentation
- lactobacillus
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Classifications
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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
- C12N1/205—Bacterial isolates
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C19/00—Cheese; Cheese preparations; Making thereof
- A23C19/06—Treating cheese curd after whey separation; Products obtained thereby
- A23C19/061—Addition of, or treatment with, microorganisms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
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- A23C9/127—Fermented milk preparations; Treatment using microorganisms or enzymes using microorganisms of the genus lactobacteriaceae and other microorganisms or enzymes, e.g. kefir, koumiss
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- A23L13/45—Addition of, or treatment with, microorganisms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- 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
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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
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
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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
- C12N1/02—Separating microorganisms from their culture media
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- 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
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
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Abstract
The present invention provides a kind of Lactobacillus plantarum A11（Lactobacillus plantarum）China Committee for Culture Collection of Microorganisms's common micro-organisms center is preserved on June 2nd, 2017, preserving number is CGMCC No.14216.
Description
Technical field
The present invention relates to a kind of Lactobacillus plantarum and its application and preparation method, belong to food microorganisms screening with applying skill
Art field.
Background technology
Lactic acid bacteria (Lactic acid bacteria LAB) is that a class can produce a class of lactic acid using carbohydrate
The general name of bacterium.Lactic acid bacteria is gram-positive cocci or bacillus, belongs to anaerobism or facultative anaerobic bacteria.In lactic acid bacteria fermentation process
In, volatile acid or gas are produced sometimes, the food such as salted vegetables and pickles are just made of it early in thousands of years of preceding ancients, near
For continuing to develop for science and technology, people gradually pay close attention to probiotic lactobacillus, and this kind of lactic acid bacteria is the bacterium beneficial to human or animal,
It can be taken as food additives, have definite health efficacy, to vivo environment（The bile of hydrochloric acid in gastric juice and enteron aisle）There is certain tolerance
Ability, can be colonized in human body intestinal canal.Mainly there is the newborn bar of lactobacillus acidophilus, plant currently as the strain of probiotic lactobacillus
Bacterium, Lactobacillus rhamnosus, lactobacillus fermenti, lactobacillus reuteri, cheese and lactobacillus paracasei, Bifidobacterium etc..
Probiotic lactobacillus can produce the metabolite of multiple beneficial, such as organic acid, polysaccharide, bacteriocin.Wherein have
Machine acid is the important substance that lactose is produced after lactobacillus-fermented, and it is not only the important indicator of acidified milk attenuation degree, is also
Influence one of key factor of acidified milk local flavor, nutrition and health.Lactic acid and the proper ratio with acetic acid in organic acid, can be with
Preferably improve enteric microorganism composition, increase intestines peristalsis, reduce corrupt substance in enteron aisle, defecation is in perfect condition,
Promote and safeguard intestinal health.Other polysaccharide has antioxidation activity, and can produce the lactic acid bacteria of exocellular polysaccharide can make probiotics
The quality of fermented product is more moist, and some polysaccharide have positive role to health, such as antitumor, blood coagulation, drop
Blood glucose, reducing blood lipid etc..
With the continuous social and economic development, the increasingly raising of living standards of the people, the benefit produced using lactobacillus-fermented
Raw based food and medicine increasingly sell well, and rapidly become the healthy food of global concern.At present, using probiotics fermention fruits and vegetables, open
Probiotics fermention beverage made of fruits or vegetables sour-sweet tasty and refreshing and with a variety of nutritional health functions are sent, are quite received by the market.Therefore, set up
Lactic acid bacteria culturers storehouse, and the research of lactic acid bacteria and exploitation Related product of the screening with beneficial function seems particularly on this basis
It is important.
The content of the invention
There is provided a kind of Lactobacillus plantarum and its application and preparation method for problem above by the present invention.
Goal of the invention is realized by following scheme：A kind of Lactobacillus plantarum（Lactobacillus plantarum）A11,
China Committee for Culture Collection of Microorganisms's common micro-organisms center is preserved on June 2nd, 2017, preserving number is CGMCC
No.14216。
Further, its colonial morphology is characterized as：Circular colonies, 3mm is wide, raised, smooth, fine and closely woven, and white is shallow once in a while
Yellow or buff；Its thalli morphology characteristic is：Gram-positive, no gemma, nose circle straight-bar bacterium is single, paired, into short chain.
Further, Lactobacillus plantarum A11（Lactobacillus plantarum A11）Organic acid is constituted after fermentation：
Oxalic acid 2.3%, tartaric acid 6.77%, malic acid 3.07%, lactic acid 77.0%, acetic acid 6.42%, citric acid 2.80%, butanedioic acid
1.65%。
A kind of application of Lactobacillus plantarum, Lactobacillus plantarum A11（Lactobacillus plantarumA11）For making
Standby food, health products, cosmetics with physiological regulatory action.
A kind of application of Lactobacillus plantarum, Lactobacillus plantarum A11（Lactobacillus plantarumA11）With it is other
Bacterial strain carry out compounding leavening is made, applied to acidified milk, fruits and vegetables, cheese, meat products food fermentation.
A kind of application of Lactobacillus plantarum, Lactobacillus plantarum A11（Lactobacillus plantarumA11）With drop
Solve cholesterol ability.
Further, the probiotic fermentation beverage with health-care efficacy is prepared into by fruits and vegetables fermentation.
A kind of application of Lactobacillus plantarum, probiotic fermentation beverage, its ferment technical recipe be：Carrot juice 25%, sugarcane
Sugar 5%, glucose 3%, K2HPO40.03%, KH2PO40.02%, MgSO40.03%, citric acid 0.1%
Further, Lactobacillus plantarum A11 is inoculated into above-mentioned formula by probiotic fermentation beverage with 2% inoculum concentration, 37 DEG C of hairs
Ferment 14h, produces lactic acid bacteria beverage made of fruits or vegetables, and wherein viable bacteria Particle density is up to 108More than CFU/ml.
A kind of method of separating plant lactobacillus, from the microbiologic population of Wenzhou Taishun mountain area traditional zymotic kimchi foodstuffs
Bacterium;The pickles sample of collection is placed in the physiological saline test tube of sterilizing, after fully shaking, 1ml is taken, 9ml is added to and goes out
In bacterium physiological saline, shaken well carries out gradient dilution 10-2,10-3,10-4 successively, respectively takes 0.1mL to be coated on the training of MRS solids
On the flat board for supporting base, 37 DEG C of culture 48h；Colonial morphology is observed, microprotrusion, milky circular colonies such as occurs and is tentatively set to mesh
Bacterial strain is marked, then picking single bacterium colony is rule repeatedly from aimed strain, until obtaining pure culture；Select the single bacterium and carry out leather orchid
Albert'stain Albert, microscopy and catalase experiment, Gram's staining is positive, and microscopy is shaft-like or spherical, and catalase is real
Test and be set to aimed strain for negative bacterial strain；The aimed strain of purifying is subjected to glycerine cryopreservation, in case follow-up preservation identification
And application.
Biomaterial preservation
Lactobacillus plantarum A11 provided by the present invention（Lactobacillus plantarum）It is preserved on June 2nd, 2017
China Committee for Culture Collection of Microorganisms's common micro-organisms center, preserving number is CGMCC No.14216, and preservation address is
Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica.
Figure of description
The standard curve of Fig. 1 cholesterol levels.
The gas chromatogram of Fig. 2 cholesterol solutions.
Embodiment
Below in conjunction with specific embodiment, the invention will be further described：
A kind of embodiment 1, Lactobacillus plantarum A11 of the invention was preserved in Chinese microorganism strain preservation pipe on June 2nd, 2017
Reason committee common micro-organisms center, preserving number is CGMCC No.14216.
A kind of Lactobacillus plantarum, its colonial morphology is characterized as：Circular colonies, 3mm is wide, raised, smooth, fine and closely woven, white,
Light yellow once in a while or buff；Its thalli morphology characteristic is：Gram-positive, no gemma, nose circle straight-bar bacterium, it is single, in pairs or
Into short chain.
A kind of Lactobacillus plantarum, Lactobacillus plantarum A11（Lactobacillus plantarum A11）16SrDNA total orders
It is classified as shown in sequence table.
A kind of Lactobacillus plantarum, Lactobacillus plantarum A11（Lactobacillus plantarum A11）Have for preparing
Food, health products or the cosmetics of physiological regulatory action.
A kind of Lactobacillus plantarum, Lactobacillus plantarum A11（Lactobacillus plantarum A11）Enter with other bacterial strains
Leavening is made in row compounding, applied to acidified milk, fruits and vegetables, cheese, meat products food fermentation.
A kind of Lactobacillus plantarum, Lactobacillus plantarum A11（Lactobacillus plantarum A11）Consolidate with degraded courage
Alcohol ability, can be prepared into the probiotic fermentation beverage with health-care efficacy by fruits and vegetables fermentation.
A kind of Lactobacillus plantarum, its ferment technical recipe be：Carrot juice 25%, sucrose 5%, glucose 3%, K2HPO4
0.03%, KH2PO4 0.02%, MgSO4 0.03%, citric acid 0.1%.
Lactobacillus plantarum A11, is inoculated into above-mentioned formula by a kind of Lactobacillus plantarum with 2% inoculum concentration, 37 DEG C of fermentations
14h, produces lactic acid bacteria beverage made of fruits or vegetables, and wherein viable bacteria Particle density reaches more than 108CFU/ml.
A kind of embodiment 2, Lactobacillus plantarum is being prepared with the application in reduction cholesterol functional product.
Embodiment 3, a kind of application of Lactobacillus plantarum in ferment agent for sour milk is prepared.
A kind of embodiment 4, Lactobacillus plantarum has oxidation resistance.
Embodiment 5, a kind of method of separating plant lactobacillus, Lactobacillus plantarum A11 is to pick up from Wenzhou Taishun mountain area farmers'
Traditional-handwork pickles in obtained by separation screening, specific separation authentication step is as follows：
The pickles sample of collection is placed in the physiological saline test tube of sterilizing, after fully shaking, 1ml is taken, 9ml sterile physiologicals are added to
In salt solution, shaken well carries out gradient dilution 10-2,10-3,10-4, respectively takes 0.1mL to be coated on MRS solid mediums successively
On flat board, 37 DEG C of culture 48h.Colonial morphology is observed, microprotrusion, milky circular colonies such as occurs and is tentatively set to object bacteria
Strain, then picking single bacterium colony is rule repeatedly from aimed strain, until obtaining pure culture.Select the single bacterium and carry out gram dye
Color, microscopy and catalase experiment, Gram's staining is positive, and microscopy is shaft-like or spherical, and catalase experiment is
Negative bacterial strain is set to aimed strain.The aimed strain of purifying is subjected to glycerine cryopreservation, in case follow-up preservation is identified and should
With.
The Lactobacillus plantarum A11's of embodiment 6 isolates and purifies and identification preservation
The Lactobacillus plantarum A11 of the present invention is separation screening gained in the traditional-handwork pickles for pick up from Wenzhou Taishun mountain area farmers',
Specific separation authentication step is as follows：
The pickles sample of collection is placed in the physiological saline test tube of sterilizing, after fully shaking, 1ml is taken, 9ml sterile physiologicals are added to
In salt solution, shaken well carries out gradient dilution 10-2,10-3,10-4, respectively takes 0.1mL to be coated on MRS solid mediums successively
On flat board, 37 DEG C of culture 48h.Colonial morphology is observed, microprotrusion, milky circular colonies such as occurs and is tentatively set to object bacteria
Strain, then picking single bacterium colony is rule repeatedly from aimed strain, until obtaining pure culture.Select the single bacterium and carry out gram dye
Color, microscopy and catalase experiment, Gram's staining is positive, and microscopy is shaft-like or spherical, and catalase experiment is
Negative bacterial strain is set to aimed strain.The aimed strain of purifying is subjected to glycerine cryopreservation, in case follow-up preservation is identified and should
With.
The aimed strain of purifying is subjected to the identification of 16S rDNA gene sequencing, concrete operations are as follows：With Ezup pillar bacteriums
Genome DNA extraction kit extracts aimed strain genomic DNA, using bacterial universal primers to 27F：5’-
AGTTTGATCMTGGCTCAG -3 ', 1492R：5 '-GGTTACCTTGTTACGACTT -3 ') carry out 16S rDNA PCR it is real
Test, after PCR reaction amplifications terminate, PCR primer is entered into row agarose gel electrophoresis, and gel imaging, it is single, clear to amplify
The bacterial strain of band is sequenced with gene sequencer, as a result as shown in SEQ ID NO.1, and BLAST sequences are carried out on NCBI websites
Row are compared, and as a result show the sequence and Lactobacillus plantarum（Lactobacillus plantarum）16S rDNA sequence homologies
Property more than 99%, with reference to the homology analysis in the morphological feature and bio-chemical characteristics of A11 bacterium, and phylogenetic tree, judge
The A11 bacterial strains of screening are Lactobacillus plantarum.
Then the aimed strain isolated and purified is subjected to API Physiology and biochemistry identifications, process is as follows：1）The preparation of inoculum：
The bacterial strain of purifying is passed on MRS broth bouillons and is inoculated in MRS agar afterwards twice, 37 DEG C of culture 24h, 5 bacterium colonies of picking
Bacteria suspension is made in the CHL culture mediums of API 50 in the consistent bacterium colony of form.2）The inoculation of reagent strip：With vaccinated API 50
CHL culture mediums add the sterile test tube for having reagent strip, and are managed with all determine of sterile mineral oil covering, 37 DEG C of culture 48h.3）Sentence
Read reagent strip：Contained Bromocresol purple turns yellow in culture（It is black from purple stain that No. 25 aesculins determine pipe）For positive findings, institute
Have and determine the biochemical collection of illustrative plates that pipe result constitutes bacterial strain, the reaction identification table of reference standard bacterial strain, it is plant to determine whether the bacterial strain
Thing lactobacillus, this result is consistent with 16S rDNA gene sequencing qualification results, final to determine that aimed strain A11 is the newborn bar of plant
Bacterium.
Strain Designation of the present invention is Lactobacillus plantarum（Lactobacillus plantarum）A11, on June 2nd, 2017
China Committee for Culture Collection of Microorganisms's common micro-organisms center is preserved in, preserving number is CGMCC No. 14216.Address
For Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, postcode 100101.
Organic acid is constituted after the Lactobacillus plantarum A11 of embodiment 7 fermentations
Lactobacillus plantarum is carried out after passage activation, MRS fluid nutrient mediums are inoculated in 2% inoculum concentration, 37 DEG C of quiescent culture 48h,
4000r/min, centrifuges 15min, collects fermented supernatant fluid, and 0.45 μm of aqueous phase membrane filtration, filtrate is analyzed for upper machine.Chromatostrip
Part mobile phase：3%CH3OH-0.5%NH4H2PO4 (pH=2.5) solution, flow velocity is 1.0mL/min, and ultraviolet detection wavelength is
210nm, sample size is 10 μ L.
Lactobacillus plantarum A11（Lactobacillus plantarum A11）Organic acid is constituted after fermentation：
Organic acid has important influence to acidified milk local flavor.The malic acid, formic acid, amber of trace are found that in traditional acidified milk
The organic acids such as amber acid, propionic acid.And significantly correlated or extremely notable phase is all presented between each organic acid content and between titratable acidity
Close.
The Lactobacillus plantarum A11 of embodiment 8 acid and bile salt tolerance experiment
Lactobacillus plantarum A11 is activated by glycerine cryopreservation tube, and picking single bacterium drops down onto 37 DEG C of constant temperature trainings in 10mL MRS fluid nutrient mediums
24h is supported, thalline is collected by centrifugation, is washed 2-3 times with sterile saline, its concentration is adjusted to 109cfu/mL, then by this bacterium solution
It is inoculated in the MRS fluid nutrient mediums that pH is 3.0,37 DEG C incubated, in the separately sampled carry out count plate of 0h and 3h, with 0h
Sample is compares, the survival rate of calculating 3h sample bacterial strains, and survival rate=（3h viable count/0h viable count）×100%
Lactobacillus plantarum A11 is activated by glycerine cryopreservation tube, and picking single bacterium drops down onto 37 DEG C of constant temperature trainings in 10mL MRS fluid nutrient mediums
24h is supported, thalline is collected by centrifugation, is washed 2-3 times with sterile saline, its concentration is adjusted to 109cfu/mL, then by this bacterium solution
It is inoculated in the MRS fluid nutrient mediums that gallbladder salinity is 0.3%, 37 DEG C incubated, in the separately sampled progress viable bacteria meter of 0h and 3h
Number, using 0h samples as control, the survival rate of calculating 3h sample bacterial strains, survival rate=（3h viable count/0h viable count）×100%
Simultaneously using lactobacillus acidophilus ATCC 43121 as control, above-mentioned acid and bile salt tolerance experiment is carried out, concrete outcome is shown in Table 1
The bacterial strain of table 1 is acidproof, bile tolerance testing result
Can lactic acid bacteria field planting survival course depends on resist internal poor environment with lactic acid bacteria, generally with the acidproof resistance to courage of lactic acid bacteria
Salt ability is used as measurement index.It is can be seen that from the data of table 1 compared with lactobacillus acidophilus ATCC 43121, we filter out
The Lactobacillus plantarum A11 come has stronger acidproof and bile tolerance ability, and especially survival rate is up under conditions of pH=3
108.56%, illustrate that Lactobacillus plantarum A11 can not only survive in hydrochloric acid in gastric juice, and slightly breed to a certain extent.This knot
Fruit demonstrates Lactobacillus plantarum A11 and possesses higher intestines and stomach survival ability, also show what the bacterial strain was developed as probiotics
Potentiality.
Removals of the Lactobacillus plantarum A11 of embodiment 9 to cholesterol is tested
The drafting of standard curve：The accurate cholesterol titer for preparing 0.2mg/mL, take 1 respectively, 2,3,4,5mL is in 10mL capacity
In bottle, isopropanol constant volume is used.Then Agilent GC HP-5 (30m × 0.32mm × 0.25 μm) are used, are detected, column temperature：
Initial temperature is 100 DEG C, keeps 1min, and 300 DEG C are warming up to 20 DEG C/min speed, keeps 10min, 250 DEG C of injector temperature,
300 DEG C of detector temperature, sample size 1 μ L, each sample feeding three times writes down peak area, using concentration as abscissa, peak area is
Ordinate, makes standard curve.
The preparation of sample：Lactobacillus plantarum A11 is activated by glycerine cryopreservation tube, and picking single bacterium drops down onto 5mL MRS Liquid Cultures
37 DEG C of incubated 24h in base, pass on 37 DEG C of incubated 24h into 10ml MRS fluid nutrient mediums, so by 2% inoculum concentration
The seed liquor that this makes is inoculated in the MRS fluid nutrient mediums containing 0.1mg/mL cholesterol by 2% inoculum concentration afterwards, 37 DEG C
Incubated 24h
Cholesterol is extracted and determined：Zymotic fluid 5000rpm is centrifuged into 15min, takes supernatant 1mL in screw-cap test tube, plus 10% and enter hydrogen-oxygen
Change potassium methanol solution 2mL（w/v）, test tube is screwed, in 75 DEG C of water-bath saponification 50min, therebetween every concussion in 10 minutes once, saponification
After the completion of, cooling adds 2mL water, 4mL n-hexanes（Repeat extraction once）, whirlpool concussion 2min, standing 10min are layered, taken
Upper strata n-hexane layer merges the n-hexane common 8mL of layer extracted twice, nitrogen drying, the dissolving of 1mL isopropanols, mistake in new test tube
Gas chromatograph is injected after 0.45 μm of filter membrane, peak area is determined, the concentration of cholesterol in sample solution, root is obtained by standard curve
It is qualitative according to retention time, quantified by external standard method.Wherein the MRS fluid nutrient mediums containing 0.1mg/mL cholesterol for not connecing bacterial strain are made
For blank control.
Simultaneously using lactobacillus acidophilus ATCC 43121 as control, above-mentioned cholesterol degradation experiment is carried out, concrete outcome is shown in
Table 2
The bacterial strain norcholesterol testing result of table 2
It can be seen that from the data of table 2：Compared with lactobacillus acidophilus ATCC 43121, the Lactobacillus plantarum that we screen
A11 has preferable cholesterol degradation ability, and can ferment the probiotics fermention drink being prepared into health-care efficacy using fruits and vegetables
Material.
The Lactobacillus plantarum A11 of embodiment 10 anti-oxidant experiment
Lactobacillus plantarum A11 after activation is passed on 2 times is inoculated into 10mL MRS fluid nutrient mediums by 2% inoculum concentration, 37 DEG C
Incubated 18h, 4000r/min, centrifuge 15min, collect fermented supernatant fluid and thalline.With the phosphate buffer solution weight of pH=7.4
Outstanding thalline, then extracts the cell-free extract of thalline with ultrasonic fragmentation.
Lactobacillus plantarum A11 fermented supernatant fluid and the antioxidation activity of cell-free extract are determined using DPPH methods：Respectively
The fermented supernatant fluid and cell-free extract of 2mL thalline are taken, the 0.2mM DPPH solution that 2mL95% ethanol is prepared is separately added into,
Shake up and stand 30 min of reaction at rear room temperature dark, its absorbance at 517nm is determined with ultraviolet specrophotometer.
DPPH clearance rates (%)=[1 (Asample-Ablank)/Acontrol] × 100
Asample:The absorbance of sample and DPPH mixed liquors；
Ablank:The absorbance of sample and 95% alcohol mixeding liquid；
Acontrol:The solvent of sample dissolution and the absorbance of DPPH mixed liquors；
The anti-oxidant testing result of the bacterial strain of table 3
As a result show：Lactobacillus plantarum A11 fermented supernatant fluid and cell-free extract has stronger oxidation resistance, to DPPH
Radical scavenging activity is respectively 96.95% and 64.21%, and fermented supernatant fluid wants star to be higher than nothing to DPPH radicals scavenging effects
Cell extract.Illustrate that the metabolite such as polysaccharide that Lactobacillus plantarum A11 fermentation process is produced is played DPPH free radicals
Prior effect, with stronger oxidation resistance.
The Lactobacillus plantarum A11 of embodiment 11 prepares probiotic fermentation beverage
The preparation of Lactobacillus plantarum A11 seed liquors：Lactobacillus plantarum A11 after activation passage is inoculated into 10mL by 2% inoculum concentration
MRS fluid nutrient mediums in, 37 DEG C of incubated 48h, 4000r/min, centrifuge 15min, collect thalline, with sterile water washing 2-
3 times, its concentration is adjusted to 109cfu/mL.
Configure liquid fermentation medium：Carrot is cleaned, stripping and slicing, the distilled water of the quality such as addition is beaten into beater
Carrot juice, the dispensing in formula is added in carrot juice so that carrot juice 25%, sucrose 5%, glucose 3%, K2HPO4
0.03%, KH2PO4 0.02%, MgSO4 0.03%, citric acid 0.1%, shearing, homogeneous after stirring, 121 DEG C of sterilizing 30min,
Cold cut is to 37 DEG C or so.
2% Lactobacillus plantarum A11 seed liquor is inoculated into liquid fermentation medium, 37 DEG C of fermentation 14h are put into 4 DEG C
Refrigerator 8h, taste allotment is carried out with sodium citrate and sucrose, you can obtain tasty, and nutritious, natural carrot taste is strong
And the probiotic fermentation beverage with good healthcare function.
Although the present invention is described by reference to preferred embodiment, those of ordinary skill in the art should
Work as understanding, the description of above-described embodiment can be not limited to, in the range of claims, can make each in form and details
Plant change.
Sequence table
<110>The ring food limited company of Zhejiang one
<120>A kind of Lactobacillus plantarum A11 and its application
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 1428
<212> DNA
<213>Lactobacillus plantarum A11（Lactobacillus plantarum A11）
<400> 1
gcaagtcgaa cgaactctgg tattgattgg tgcttgcatc atgatttaca tttgagtgag 60
tggcgaactg gtgagtaaca cgtgggaaac ctgcccagaa gcgggggata acacctggaa 120
acagatgcta ataccgcata acaacttgga ccgcatggtc cgagtttgaa agatggcttc 180
ggctatcact tttggatggt cccgcggcgt attagctaga tggtggggta acggctcacc 240
atggcaatga tacgtagccg acctgagagg gtaatcggcc acattgggac tgagacacgg 300
cccaaactcc tacgggaggc agcagtaggg aatcttccac aatggacgaa agtctgatgg 360
agcaacgccg cgtgagtgaa gaagggtttc ggctcgtaaa actctgttgt taaagaagaa 420
catatctgag agtaactgtt caggtattga cggtatttaa ccagaaagcc acggctaact 480
acgtgccagc agccgcggta atacgtaggt ggcaagcgtt gtccggattt attgggcgta 540
aagcgagcgc aggcggtttt ttaagtctga tgtgaaagcc ttcggctcaa ccgaagaagt 600
gcatcggaaa ctgggaaact tgagtgcaga agaggacagt ggaactccat gtgtagcggt 660
gaaatgcgta gatatatgga agaacaccag tggcgaaggc ggctgtctgg tctgtaactg 720
acgctgaggc tcgaaagtat gggtagcaaa caggattaga taccctggta gtccataccg 780
taaacgatga atgctaagtg ttggagggtt tccgcccttc agtgctgcag ctaacgcatt 840
aagcattccg cctggggagt acggccgcaa ggctgaaact caaaggaatt gacgggggcc 900
cgcacaagcg gtggagcatg tggtttaatt cgaagctacg cgaagaacct taccaggtct 960
tgacatacta tgcaaatcta agagattaga cgttcccttc ggggacatgg atacaggtgg 1020
tgcatggttg tcgtcagctc gtgtcgtgag atgttgggtt aagtcccgca acgagcgcaa 1080
cccttattat cagttgccag cattaagttg ggcactctgg tgagactgcc ggtgacaaac 1140
cggaggaagg tggggatgac gtcaaatcat catgcccctt atgacctggg ctacacacgt 1200
gctacaatgg atggtacaac gagttgcgaa ctcgcgagag taagctaatc tcttaaagcc 1260
attctcagtt cggattgtag gctgcaactc gcctacatga agtcggaatc gctagtaatc 1320
gcggatcagc atgccgcggt gaatacgttc ccgggccttg tacacaccgc ccgtcacacc 1380
atgagagttt gtaacaccca aagtcggtgg ggtaaccttt aggaacca 1428
Claims (9)
1. a kind of Lactobacillus plantarum（Lactobacillus plantarum）A11, Chinese micro- life is preserved on June 2nd, 2017
Thing culture presevation administration committee common micro-organisms center, preserving number is CGMCC No.14216.
2. Lactobacillus plantarum according to claim 1, it is characterised in that：Its colonial morphology is characterized as：Circular colonies,
3mm is wide, raised, smooth, fine and closely woven, white, light yellow once in a while or buff；Its thalli morphology characteristic is：Gram-positive, no bud
Spore, nose circle straight-bar bacterium is single, paired, into short chain.
3. Lactobacillus plantarum according to claim 1, it is characterised in that：Lactobacillus plantarum A11（Lactobacillus
plantarum A11）Organic acid is constituted after fermentation：Oxalic acid 2.3%, tartaric acid 6.77%, malic acid 3.07%, lactic acid 77.0%, second
Acid 6.42%, citric acid 2.80%, butanedioic acid 1.65%.
4. a kind of application of Lactobacillus plantarum according to claim 1, it is characterised in that：Lactobacillus plantarum A11
（Lactobacillus plantarumA11）For preparing food, health products, cosmetics with physiological regulatory action.
5. a kind of application of Lactobacillus plantarum according to claim 1, it is characterised in that：Lactobacillus plantarum A11
（Lactobacillus plantarumA11）With other bacterial strains carry out compounding leavening is made, applied to acidified milk, fruits and vegetables,
The fermentation of cheese, meat products food.
6. a kind of application of Lactobacillus plantarum according to claim 1, it is characterised in that：Lactobacillus plantarum A11
（Lactobacillus plantarumA11）With cholesterol degradation ability.
7. the application of Lactobacillus plantarum according to claim 6, it is characterised in that：It is prepared into by fruits and vegetables fermentation with guarantor
The probiotic fermentation beverage of strong effect.
8. a kind of application of Lactobacillus plantarum according to claim 7, it is characterised in that：Described probiotics fermention drink
Expect, its technical recipe fermented is：Carrot juice 25%, sucrose 5%, glucose 3%, K2HPO40.03%, KH2PO40.02%,
MgSO40.03%, citric acid 0.1%
The application of Lactobacillus plantarum according to claim 7 or 8, it is characterised in that：Probiotic fermentation beverage connects with 2%
Lactobacillus plantarum A11 is inoculated into above-mentioned formula by the amount of kind, 37 DEG C of fermentation 14h, produces lactic acid bacteria beverage made of fruits or vegetables, wherein viable count
Concentration is up to 108More than CFU/ml.
9. a kind of method for separating the Lactobacillus plantarum described in claim 1, it is characterised in that：From Wenzhou Taishun mountain area, tradition is sent out
Bacterium in the microbiologic population of fermented pickled vegetable food;The pickles sample of collection is placed in the physiological saline test tube of sterilizing, filled
Divide after concussion, take 1ml, be added in 9ml sterile salines, shaken well, gradient dilution 10-2,10-3,10-4 are carried out successively,
0.1mL is respectively taken to be coated on the flat board of MRS solid mediums, 37 DEG C of culture 48h；Colonial morphology is observed, microprotrusion, breast such as occurs
The circular colonies of white are tentatively set to aimed strain, and then picking single bacterium colony is rule repeatedly from aimed strain, until obtaining pure
Culture；Select the single bacterium and carry out Gram's staining, microscopy and catalase experiment, Gram's staining is positive, microscopy
To be shaft-like or spherical, catalase experiment is set to aimed strain for negative bacterial strain；The aimed strain of purifying is subjected to glycerine
Cryopreservation, in case follow-up preservation identification and application.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108977396A (en) * | 2018-08-20 | 2018-12-11 | 江南大学 | A kind of lactobacillus plantarum of high-yield lactic acid and its application |
| CN109170437A (en) * | 2018-10-12 | 2019-01-11 | 中国海洋大学 | A kind of viable type banana carrot composite juice beverage and preparation method thereof |
| CN109234189A (en) * | 2018-08-17 | 2019-01-18 | 甘肃普诺贝康生物科技有限责任公司 | One plant of lactobacillus plantarum strain BX62 and its application with oxidation resistance |
| CN110272837A (en) * | 2018-12-13 | 2019-09-24 | 山东理工大学 | A kind of lactobacillus plantarum S3-10 and its screening technique and application |
| CN110903996A (en) * | 2019-10-15 | 2020-03-24 | 江南大学 | Method for preparing yogurt by lactobacillus plantarum with high short-chain fatty acid yield |
| CN112195139A (en) * | 2020-11-10 | 2021-01-08 | 广西壮族自治区农业科学院 | Lactobacillus plantarum strain LDVS007 and application thereof |
| CN113308416A (en) * | 2021-07-20 | 2021-08-27 | 四川大学 | Lactobacillus plantarum capable of inhibiting kidney stone formation and application thereof |
| CN115232768A (en) * | 2022-07-22 | 2022-10-25 | 江南大学 | Lactobacillus paracasei JN-8 and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101864375A (en) * | 2010-01-15 | 2010-10-20 | 吉林省农业科学院 | Lactobacillu plantarurn with function of reducing cholesterol and application thereof |
| CN105725215A (en) * | 2016-03-28 | 2016-07-06 | 济南康多宝生物技术有限公司 | Lactobacillus oral solution and preparation method thereof |
| CN105779343A (en) * | 2016-03-28 | 2016-07-20 | 济南康多宝生物技术有限公司 | Lactobacillus plantarum capable of effectively removing cholesterol and free radicals and application thereof |
-
2017
- 2017-07-17 CN CN201710580930.2A patent/CN107227278A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101864375A (en) * | 2010-01-15 | 2010-10-20 | 吉林省农业科学院 | Lactobacillu plantarurn with function of reducing cholesterol and application thereof |
| CN105725215A (en) * | 2016-03-28 | 2016-07-06 | 济南康多宝生物技术有限公司 | Lactobacillus oral solution and preparation method thereof |
| CN105779343A (en) * | 2016-03-28 | 2016-07-20 | 济南康多宝生物技术有限公司 | Lactobacillus plantarum capable of effectively removing cholesterol and free radicals and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| JIAN ZHANG等: "Potential Probiotic Characterization of Lactobacillus plantarum Strains Isolated from Inner Mongolia "Hurood" Cheese", 《J. MICROBIOL. BIOTECHNOL.》 * |
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| CN109234189A (en) * | 2018-08-17 | 2019-01-18 | 甘肃普诺贝康生物科技有限责任公司 | One plant of lactobacillus plantarum strain BX62 and its application with oxidation resistance |
| CN108977396A (en) * | 2018-08-20 | 2018-12-11 | 江南大学 | A kind of lactobacillus plantarum of high-yield lactic acid and its application |
| CN108977396B (en) * | 2018-08-20 | 2020-08-04 | 江南大学 | Lactobacillus plantarum for high yield of lactic acid and application thereof |
| CN109170437A (en) * | 2018-10-12 | 2019-01-11 | 中国海洋大学 | A kind of viable type banana carrot composite juice beverage and preparation method thereof |
| CN110272837A (en) * | 2018-12-13 | 2019-09-24 | 山东理工大学 | A kind of lactobacillus plantarum S3-10 and its screening technique and application |
| CN110903996A (en) * | 2019-10-15 | 2020-03-24 | 江南大学 | Method for preparing yogurt by lactobacillus plantarum with high short-chain fatty acid yield |
| CN112195139A (en) * | 2020-11-10 | 2021-01-08 | 广西壮族自治区农业科学院 | Lactobacillus plantarum strain LDVS007 and application thereof |
| CN113308416A (en) * | 2021-07-20 | 2021-08-27 | 四川大学 | Lactobacillus plantarum capable of inhibiting kidney stone formation and application thereof |
| CN113308416B (en) * | 2021-07-20 | 2023-02-24 | 四川大学 | Lactobacillus plantarum capable of inhibiting kidney stone formation and application thereof |
| CN115232768A (en) * | 2022-07-22 | 2022-10-25 | 江南大学 | Lactobacillus paracasei JN-8 and application thereof |
| CN115232768B (en) * | 2022-07-22 | 2023-04-14 | 江南大学 | Lactobacillus paracasei JN-8 and application thereof |
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