CN105062918B - One lactobacillus plantarum and its application - Google Patents
One lactobacillus plantarum and its application Download PDFInfo
- Publication number
- CN105062918B CN105062918B CN201510464648.9A CN201510464648A CN105062918B CN 105062918 B CN105062918 B CN 105062918B CN 201510464648 A CN201510464648 A CN 201510464648A CN 105062918 B CN105062918 B CN 105062918B
- Authority
- CN
- China
- Prior art keywords
- lactobacillus plantarum
- lactobacillus
- application
- free radical
- product
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention discloses a lactobacillus plantarum and its applications.Lactobacillus plantarum (Lactobacillus plantarum) NC301 provided by the invention, preservation registration number are CGMCC NO.11013.Lactobacillus plantarum NC301 also belongs to protection scope of the present invention in the application prepared in the product for adjusting intestinal flora.The present invention also protects lactobacillus plantarum NC301 preparing the application in antioxidant.Bacterial strain provided by the invention is one plant of Lactic Acid Bacteria new strains with potential probiotic properties, can be used as the probiotics for adjusting intestinal flora, improvement function of intestinal canal applied to food, health care product and medicine field, application prospect is very wide.
Description
Technical field
The invention belongs to microorganisms technical fields, and in particular to a lactobacillus plantarum and its application.
Background technique
Chinese tradition fermented food is with a long history, unique flavor, by the favor of consumer.Traditional fermented food includes hair
Ferment vegetables, fermentation meat product, fermented dairy product, white wine and fermented seasonings etc. are the important components of China's food industry.
Lactic acid bacteria in traditional fermented food not only contributes to improve flavor, mouthfeel and the quality etc. of fermented food, and some lactic acid bacterias are also
With excellent probiotic properties.
The superior geographical environment in Changbaishan area creates local natural resources abundant, forms very characteristic fermentation
Food, such as pickles, salted vegetables.By long-term natural domestication, it is benefit that lactic acid bacteria, which becomes the dominant microflora in these traditional cuisines,
The good source of raw bacterium.
Summary of the invention
The object of the present invention is to provide a lactobacillus plantarum and its applications.
Lactobacillus plantarum (Lactobacillus plantarum) NC301 provided by the invention, in 06 month 2015 23
Day is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (abbreviation CGMCC, address are as follows: court of Beijing
The institute 3 of positive area's North Star West Road 1, Institute of Microorganism, Academia Sinica), preservation registration number is CGMCC NO.11013.Plant
Lactobacillus (Lactobacillus plantarum) NC301CGMCC NO.11013, abbreviation lactobacillus plantarum NC301.
Lactobacillus plantarum NC301 also belongs to protection of the invention in the application prepared in the product for adjusting intestinal flora
Range.The function of " for the adjusting the product of intestinal flora " be (a) and or (b) and or (c) and or (d): (a) promote
Lactobacillus increases；(b) Bifidobacterium is promoted to increase；(c) enterococcus is promoted to reduce；(d) enterobacteria is promoted to reduce.
The present invention also protects a kind of for adjusting the product of intestinal flora, and active constituent is lactobacillus plantarum NC301.Institute
State function " for adjusting the product of intestinal flora " be (a) and or (b) and or (c) and or (d): (a) lactobacillus is promoted to increase
Add；(b) Bifidobacterium is promoted to increase；(c) enterococcus is promoted to reduce；(d) enterobacteria is promoted to reduce.
The present invention also protects lactobacillus plantarum NC301 preparing the application in antioxidant.The function of the antioxidant
To remove free radical.The free radical be hydroxy radical and or DPPH free radical and or ultra-oxygen anion free radical.
The present invention also protects a kind of antioxidant, and active constituent is lactobacillus plantarum NC301.The function of the antioxidant
It can be removing free radical.The free radical be hydroxy radical and or DPPH free radical and or ultra-oxygen anion free radical.
The present invention also protects lactobacillus plantarum NC301 preparing the application in food, health care product or drug.The food tool
Body can be Yoghourt.
The present invention isolated lactobacillus plantarum from traditional zymotic Tian jin cabbage pickled in sweet and sour passes through external acid tolerance and cholate
The test such as tolerance, Adhering capacity, antioxidation and improvement function of intestinal canal and adjusting intestinal flora, it was confirmed that the bacterial strain is one plant
Lactic Acid Bacteria new strains with potential probiotic properties, the probiotics that can be used as and adjust intestinal flora, improve function of intestinal canal
Applied to food, health care product and medicine field, application prospect is very wide.
Detailed description of the invention
Fig. 1 is the photo of lactobacillus plantarum NC301 under an optical microscope.
Fig. 2 is photo of the lactobacillus plantarum NC301 under transmission electron microscope.
Fig. 3 is regulating power of the lactobacillus plantarum NC301 to enteron aisle flora.
Specific embodiment
Embodiment below facilitates a better understanding of the present invention, but does not limit the present invention.Experiment in following embodiments
Method is unless otherwise specified conventional method.Test material as used in the following examples is unless otherwise specified certainly
What routine biochemistry reagent shop was commercially available.Quantitative test in following embodiment is respectively provided with three repeated experiments, as a result makes even
Mean value.Bilein: the big biological medicine company limited liability company of Henan benefit, product number P333004, No. CAS is 8008-63-7.
Caco-2 cell: the American Type Culture Collection committee of Chinese Academy of Sciences cell bank.
BCP fluid nutrient medium: yeast extract 2.5g/L, peptone 5g/L, glucose 5g/L, bromocresol purple 0.04g/L, solvent
For water, pH7.0；121 DEG C of sterilizing 15min.Agar is added on the basis of BCP fluid nutrient medium to 15g/L, as BCP solid
Culture medium.
MRS fluid nutrient medium: peptone 10g/L, beef extract 10g/L, yeast extract 5g/L, KH2PO42g/L, sodium acetate 5g/
L, sodium citrate 5g/L, MgSO4·7H2O 0.2g/L、MnSO4·4H2O 0.05g/L, Tween 80 1mL, glucose 20g/L, it is molten
Agent is water, pH6.6；121 DEG C of sterilizing 15min.It is solid to 15g/L, as MRS that agar is added on the basis of MRS fluid nutrient medium
Body culture medium
The preparation method of the PBS buffer solution of pH7.2: NaCl 8g, KCl 0.2g, Na2HPO41.42g K2HPO40.27g,
Water is added to be settled to 1L, concentrated hydrochloric acid tune pH7.2.
The preparation method of the PBS buffer solution of pH7.4: NaCl 8g, KCl 0.2g, Na2HPO41.42g K2HPO40.27g,
Water is added to be settled to 1L, concentrated hydrochloric acid tune pH7.4.
Lactobacillus rhamnosus LGG is one plant of existing probiotics, is indicated with LGG.Refer to the text of Lactobacillus rhamnosus LGG
It offers: Zhang Qilin, Ma Chunli, Li Aili, Cao Nan, Song Lanlan, application of the Lactobacillus rhamnosus LGG in Cottage cheese, " food
Product industry " 04 phase in 2015.
The separation and identification of embodiment 1, bacterial strain
Sample for strain isolation is the traditional zymotic Tian jin cabbage pickled in sweet and sour of Yanbian of China.
It crosses repeatedly culture through BCP solid medium tablets, the single colonie that picking produces yellow circle is inoculated in MRS solid medium
Continue to cultivate on plate, crosses repeatedly culture purified through MRS solid medium tablets, obtain the bacterial strain of more plants of pure cultures.Wherein
One plant is named as NC301 bacterial strain.
Two, the identification of bacterial strain
The photo of NC301 bacterial strain under an optical microscope is shown in Fig. 1.Somatic cells are in nose circle direct rod shape, it is most of for it is single,
It in pairs or is in cell chain.
Photo of the NC301 bacterial strain under transmission electron microscope is shown in Fig. 2.Elongated rod shape, cell wall, cell membrane are complete, and cytoplasm is uniform.
NC301 bacterial strain is no motion of bacillus.
37~42 DEG C of NC301 bacterial strain optimum growth temperature, appropriate pH are 5.0~7.0.
The physiological and biochemical property of NC301 bacterial strain: Gram-positive, negative catalase can give birth in 15 DEG C and 45 DEG C
It is long, it is resistant to 6.5%NaCl, does not hydrolyze starch, do not liquefy gelatin, does not generate hydrogen sulfide, and glucose fermentation produces acid but do not produce gas, biphenyl
Amine test is negative, and indole test is negative, and voges-Proskauer test is positive.
API 50CH (French Mei Liai company) qualification result shows that NC301 bacterial strain is lactobacillus plantarum.NC301 bacterial strain energy
It utilizes: ribose, galactolipin, glucose, fructose, mannose, mannitol, sorbierite, amarogentin, arbutin, N- acetyl-grape
Osamine, seven leaf-alcohols, salicin, cellobiose, maltose, lactose, melibiose, sucrose, trehalose, melezitose, raffinose, Mang ox
Sugar, D-arabinose alcohol, gluconate.NC301 bacterial strain cannot utilize: arabinose, xylose, adonite, sorbose, sandlwood
Sugar, dulcitol, inositol, Alpha-Methyl-D-Glucose glycosides, Alpha-Methyl-D-MANNOSE glycosides, synanthrin, starch, glycogen, xylitol, D- pine
Disaccharides, D-Tag, D- meter threose, rock sugar, L-arabinose alcohol, 2- keto-D-gluconate salt, 5- keto-D-gluconate salt.
The 16s rDNA partial sequence of NC301 bacterial strain is as shown in sequence 1 in sequence table.
The above qualification result shows that NC301 bacterial strain belongs to lactobacillus plantarum (Lactobacillus plantarum).
Three, the preservation of bacterial strain
Lactobacillus plantarum (Lactobacillus plantarum) NC301 provided by the invention, in 06 month 2015 23
Day is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (abbreviation CGMCC, address are as follows: court of Beijing
The institute 3 of positive area's North Star West Road 1, Institute of Microorganism, Academia Sinica), preservation registration number is CGMCC NO.11013.Plant
Lactobacillus (Lactobacillus plantarum) NC301CGMCC NO.11013, abbreviation lactobacillus plantarum NC301 is used
NC301 is indicated.
Embodiment 2, the measurement of the tolerance of lactobacillus plantarum NC301
1, lactobacillus plantarum NC301 is seeded to MRS fluid nutrient medium, obtain bacteria suspension (bacteria concentration in bacteria suspension:
log10Cfu/ml=8.57 ± 0.17), bacteria suspension is divided into 4 groups (every group of 5 reprocessings), is proceeded as follows respectively:
First group: being 2.0,37 DEG C of stationary culture 3h with hydrochloric acid tune pH；
Second group: being 3.0,37 DEG C of stationary culture 3h with hydrochloric acid tune pH；
Third group: bilein is added and makes its concentration 0.3g/100ml, 37 DEG C of stationary culture 3h；
4th group: bilein is added and makes its concentration 0.6g/100ml, 37 DEG C of stationary culture 4h；
After completing aforesaid operations, 50 μ l are taken to be coated with MRS solid medium tablets, bacterium colony counts after 37 DEG C of stationary culture 48h.
It the results are shown in Table 1.Lactobacillus plantarum NC301 has certain tolerance, survival rate 20.63% to pH 2.0.It plants
Object lactobacillus NC301 has good tolerance effect to pH3.0, and survival rate is still above 80%.NC301 pairs of lactobacillus plantarum
The bovine bile of 0.3g/100ml and 0.6g/100ml all has stronger tolerance.
2, Lactobacillus rhamnosus LGG is seeded to MRS fluid nutrient medium, obtain bacteria suspension (bacteria concentration in bacteria suspension:
log10Cfu/ml=8.25 ± 0.37), bacteria suspension is divided into 4 groups (every group of 5 reprocessings), is proceeded as follows respectively:
First group: being 2.0,37 DEG C of stationary culture 3h with hydrochloric acid tune pH；
Second group: being 3.0,37 DEG C of stationary culture 3h with hydrochloric acid tune pH；
Third group: bilein is added and makes its concentration 0.3g/100ml, 37 DEG C of stationary culture 3h；
4th group: bilein is added and makes its concentration 0.6g/100ml, 37 DEG C of stationary culture 4h；
After completing aforesaid operations, 50 μ l are taken to be coated with MRS solid medium tablets, bacterium colony counts after 37 DEG C of stationary culture 48h.
It the results are shown in Table 1.
Tolerance of 1 bacterial strain of table to acid and cholate
The Adhering capacity of embodiment 3, lactobacillus plantarum NC301
Surface hydrophobic is related with the Adhering capacity of bacterial strain.The hydrophobic property on bacterial strain surface is that influence bacterial strain gut epithelium is thin
One of an important factor for born of the same parents' Adhering capacity.
One, hydrophobicity measures
1, strain to be tested is taken, with the PBS buffer solution suspension thalline of pH7.2, obtains OD600nm=0.4 bacteria suspension.
2, the bacteria suspension for taking 3mL step 1 to obtain is mixed with 1mL organic reagent, and whirlpool shakes 30s, is then stored at room temperature
30min, phase of fetching water.
3, the water phase for taking step 2 to obtain, the absorbance value measured under 600nm (are denoted as Ax)。
The hydrophobic ability of bacterial strain (%)=[1- (Ax/A0)] × 100%；A0=0.4.
Strain to be tested is lactobacillus plantarum NC301 or Lactobacillus rhamnosus LGG.
Organic reagent is dimethylbenzene, chloroform or ethyl acetate.
It carries out repeating to test three times, results are averaged.
It the results are shown in Table 2.Compared with Lactobacillus rhamnosus LGG, lactobacillus plantarum NC301 paraxylene, chloroform and acetic acid second
Ester all has stronger hydrophobic ability, shows that its surface has extremely strong electron donor characteristic.
Two, Caco-2 cell adhesion is tested
Strain to be tested is lactobacillus plantarum NC301 or Lactobacillus rhamnosus LGG.
1, in 24 porocyte culture plates, using containing 10% (volume ratio) newborn bovine serum, 100U/mL penicillin and
The DMEM culture solution culture Caco-2 cell of 100U/mL streptomysin is counted, every hole cell number when long adherent to cell monolayer
Mesh is 3.5 × 105It is a.
2, strain to be tested is taken, with the PBS buffer solution suspension thalline of pH7.2, obtaining bacteria concentration is 3.4 × 109Cfu/m1's
Bacteria suspension.
3, packet transaction
Test group: 24 porocyte culture plates for taking step 1 to obtain, every hole be added the bacteria suspension that 0.5mL step 2 obtains and
0.5mL fresh DMEM culture solution, sets 37 DEG C, 5%CO2, 90% humidity environment in stationary incubation 2h, then with pH7.4
PBS buffer solution rinsing cell 5 times (remove the bacterium that do not stick).
Control group: the bacteria suspension for replacing step 2 to obtain with the PBS buffer solution of isometric pH7.2, other same test groups.
Every group of setting three reprocessings.
4, after completing step 3,24 orifice plate is taken, fixes 30min, Gram's staining, microscopy with 0.4% paraformaldehyde.
5, after completing step 4,24 orifice plate is taken, every hole is added 0.25% pancreatin of 0.2mL and digests 10min, then every hole
DMEM culture solution of the 0.125mL containing 20% (volume ratio) serum is added to terminate reaction, then 0.175mL 0.1% is added in every hole
(volume ratio) Triton X-100 aqueous solution, is stored at room temperature 10min (with lytic cell), then carries out plate count.
Bacterial population × 100% added in sum/every hole of bacterium is attached in adherence rate (%)=every hole.
It the results are shown in Table 2.Compared with Lactobacillus rhamnosus LGG, lactobacillus plantarum NC301 Adhering capacity is stronger, shows plant
Lactobacillus NC301 has stronger Adhering capacity to enterocyte.
2 surface hydrophobic of table and Caco-2 cell adhesion ability
Embodiment 4, lactobacillus plantarum NC301 are to the adjustment effect of intestinal flora
Intestinal flora regulating power is the important and basic function of probiotics.The realization of probiotics function in addition to bacterial strain from
The functionality that body has is outer, mainly completes by adjusting the flora of host intestine, and intestinal flora is often compared to the organ of human body,
With the synthesis and metabolism for promoting nutrient, activation and adjusting immune system mitigate enteron aisle and do not accommodate diarrhea, prevent and reduce
The functions such as the generation and body self-regeneration of enteric infection.
Kunming mice (4 week old, male, 18-22g), is purchased from the high-new medical faunae center in Changchun.
Kunming mice is randomly divided into 2 groups (every group 10), is handled as follows respectively:
Test group: (volume 0.4mL contains 4 × 10 to lactobacillus plantarum bacterium solution of daily stomach-filling8Cfu lactobacillus plantarum
NC301, solvent are the PBS buffer solution of pH7.2), continuous gavage 14 days (test the 1st to 14 day), then normal raising；
Control group: the PBS buffer solution (volume 0.4mL) of pH7.2 of daily stomach-filling, continuous gavage (test the 1st in 14 days
To 14 days), then normal raising.
Respectively at test the 0th day, the 7th day, the 14th day, the 16th day, the excrement of the 21st day acquisition mouse, detection wherein newborn bar
The quantity of bacterium, Bifidobacterium, enterococcus and enterobacteria.
Lactobacillus is detected in such a way that selection plate culture is in conjunction with morphologic observation.What is used selects plate for lactobacillus
Agar medium (LBS), the preparation method is as follows: pancreas casein peptone 10g, yeast extract 5g, potassium dihydrogen phosphate 6g, ferrous sulfate
0.034g, magnesium sulfate 0.575g, glucose 20g, sodium acetate 25g, ammonium citrate 2g, manganese sulfate 0.12g, Tween 80 1mL, agar
12g, hydrochloric acid tune pH value to 5.5, heating stirring are dissolved in 1000mL distilled water, 115 DEG C of high pressure sterilization 15min.Condition of culture:
37 DEG C, 48h.The morphological feature of lactobacillus: white or milky, the sporeless bacterium that Gram's staining is positive.
Bifidobacterium is detected in such a way that selection plate culture is in conjunction with morphologic observation.What is used selects plate for bifid
Baccilus medium (MRS+NNLP), to contain acidum nalidixicum 15mg/L, neomycinsulphate 100mg/L, paromomycin sulfate 200mg/
L, the MRS solid medium of lithium chloride 3g/L, L-cysteine hydrochloride 500mg/L.Condition of culture: 37 DEG C of anaerobism, 48h.It is double
The morphological feature of discrimination bacillus: micro white, surface is smooth, protrusion, the forked or rodlike bacterium colony that Gram's staining is positive.
Enterobacteria is detected in such a way that selection plate culture is in conjunction with morphologic observation.What is used selects plate for enterobacteriaceae
Count Agar culture medium (Violet Red Bile Dextrose Agar, VRBDA), the preparation method is as follows: yeast powder 3g, egg
White peptone 7g, sodium chloride 5g, glucose 10g, cholate 1.5g, crystal violet 0.002g, dimethyl diaminophenazine chloride 0.03g, agar 12g, hydrochloric acid tune pH
Value 7.3 dissolves by heating in 1000mL distilled water, boils and do not exceed 2min, when being cooled to 50 DEG C or so, be poured into sterilized petri dishes.
Condition of culture: 37 DEG C, for 24 hours.The morphological feature of enterobacteria: lactose fermenters, the bacterium colony that Gram's staining is negative.
Enterococcus is detected in such a way that selection plate culture is in conjunction with morphologic observation.What is used selects plate for enterococcus
Agar (cholate-aesculin-Sodium azide agar) culture medium (Bile Esculin Azide Agar, BEA), preparation method is such as
Under: beef extract powder 3g, pancreas casein peptone 17g, yeast powder 5g, bilein 10g, sodium chloride 5g, aesculin 1g, ferric citrate
0.5g, sodium azide 0.25g, sodium citrate 1g, agar 13g, hydrochloric acid tune pH value 7.1 dissolve by heating in 1000mL distilled water,
121 DEG C of high pressure sterilization 15min.Condition of culture: 37 DEG C, 48h.Enterococcal morphological feature: obvious brown circle, Gram's staining are in
Positive bacterium colony.
As a result see that (filled black column represents control group to Fig. 3, and grey packed column represents test group；A: lactobacillus；B: intestines ball
Bacterium；C: Bifidobacterium；D: enterobacteria).Compared with the control group, in the excrement of test group mouse lactobacillus and Bifidobacterium quantity
It obviously increases, the quantity of enterococcus and enterobacteria reduces.The result shows that lactobacillus plantarum NC301 can promote in mouse intestinal
Lactobacillus and Bifidobacterium are increased, and enterococcus and enterobacteria are reduced.
The antioxidant activity in vitro of embodiment 5, lactobacillus plantarum NC301
One, Hydroxyl radical-scavenging ability measurement (Fenton method)
1, being suspended with the PBS buffer solution of pH7.2, (bacterium to be measured is lactobacillus plantarum NC301 or Lactobacillus rhamnosus to bacterium to be measured
LGG), obtaining bacteria concentration is 1 × 1010The bacteria suspension of cfu/mL.
2, packet transaction
Test group: taking test tube, sequentially adds 0.435mM brilliant green aqueous solution 1mL, 0.5mM ferrous sulfate aqueous solution 2mL, 3%
(volume ratio) aqueous hydrogen peroxide solution 1.5mL (Fe2+The oxidation system constituted with hydrogen peroxide is known as Fenton reagent) and 1mL step
The bacteria suspension of rapid 1 preparation, is uniformly mixed, and then 37 DEG C of waters bath with thermostatic control keep the temperature 20min, then measures supernatant at 624nm
Absorbance；
Control group first: the bacteria suspension for replacing step 1 to prepare with the PBS buffer solution of isometric pH7.2, other same test groups；
Control group second: the bacteria suspension for replacing step 1 to prepare with the PBS buffer solution of isometric pH7.2 is replaced with isometric water
Brilliant green aqueous solution replaces aqueous hydrogen peroxide solution, other same test groups with isometric water；
Every group of setting five reprocessings.
Scavenging activity (%)=[(As-Ao)/(A-Ao)] × 100% of the bacterial strain to hydroxy radical；
In formula: the absorbance value of As test group；The absorbance value of Ao expression control group first；The extinction of A expression control group second
Angle value.
Two, DPPH free radical scavenging ability measures
1, being suspended with the PBS buffer solution of pH7.2, (bacterium to be measured is lactobacillus plantarum NC301 or Lactobacillus rhamnosus to bacterium to be measured
LGG), obtaining bacteria concentration is 1 × 1010The bacteria suspension of cfu/mL.
2, packet transaction
Test group: taking 2mL 0.5mM DPPH solution, (full name of DPPH is 1,1- diphenyl -2- trinitrophenyl-hydrazine, D PPH
The solvent of solution is methanol), the bacteria suspension that 1mL step 1 obtains is added, is uniformly mixed, then room temperature avoid light place 30min, so
4 DEG C afterwards, 8000g centrifugation 10min, take supernatant, measure absorbance of the supernatant at 517nm；
Negative control group: the bacteria suspension for replacing step 1 to prepare with the PBS buffer solution of isometric pH7.2, it is other with test
Group；
Blank control group: DPPH solution, other same test groups are replaced with isometric methanol；
Every group of setting five reprocessings.
Scavenging activity (%)=[1- (A of the bacterial strain to DPPH free radicalSample-ABlank)/AControl] × 100%；
In formula: ASampleRepresent the absorbance value of test group；ABlankRepresent the absorbance value of blank control group；AControlRepresent feminine gender
The absorbance value of control group.
Three, superoxide anion Scavenging activity measures
1, being suspended with the PBS buffer solution of pH7.2, (bacterium to be measured is lactobacillus plantarum NC301 or Lactobacillus rhamnosus to bacterium to be measured
LGG), obtaining bacteria concentration is 1 × 1010The bacteria suspension of cfu/mL.
2, packet transaction
Test group: taking test tube, sequentially adds bacteria suspension, 2.8mL 3mM diethylenetriamine pentaacetic acid that 0.3mL step 1 obtains
Aqueous solution, 0.2mL 1.2mM pyrogallol aqueous solution are uniformly mixed, and then 25 DEG C of waters bath with thermostatic control keep the temperature 10min, are then measured
Absorbance at 325nm；
Control group first: pyrogallol aqueous solution, other same test groups are replaced with isometric water；
Control group second: the bacteria suspension for replacing step 1 to prepare with the PBS buffer solution of isometric pH7.2, other same test groups；
Control group third: the bacteria suspension for replacing step 1 to prepare with the PBS buffer solution of isometric pH7.2 is replaced with isometric water
Pyrogallol aqueous solution, other same test groups；
Every group of setting five reprocessings.
Scavenging activity (%)=[1- (A of the bacterial strain to superoxide anion11-A10)/(A01-A00)] × 100%
In formula: A11The absorbance value of test group；A10Represent the OD value of control group first；A01Represent the OD value of control group second；A00
Represent the OD value of control group third.
Step 1: step 2 and step 3 the results are shown in Table 3.Lactobacillus plantarum NC301 is free to hydroxy radical, DPPH
Base, ultra-oxygen anion free radical clearance rate be more than 50.0%.
3 free radical scavenging ability of table
| Bacterial strain | Hydroxy radical (%) | DPPH free radical (%) | Ultra-oxygen anion free radical (%) |
| LGG | 52.05±2.01 | 42.10±1.23 | 39.01±2.01 |
| NC301 | 62.35±1.28 | 60.14±1.54 | 54.35±1.90 |
Claims (12)
- Lactobacillus plantarum 1. (Lactobacillus plantarum) NC301, deposit number is CGMCC NO.11013.
- 2. lactobacillus plantarum described in claim 1 is preparing the application in the product for adjusting intestinal flora.
- 3. application as claimed in claim 2, it is characterised in that: the function of " for the adjusting the product of intestinal flora " is (a) and or (b) and or (c) and or (d): (a) promote lactobacillus increase；(b) Bifidobacterium is promoted to increase；(c) promote intestines ball Bacterium is reduced；(d) enterobacteria is promoted to reduce.
- 4. a kind of for adjusting the product of intestinal flora, active constituent is lactobacillus plantarum described in claim 1.
- 5. product as claimed in claim 4, it is characterised in that: the function of " for the adjusting the product of intestinal flora " is (a) and or (b) and or (c) and or (d): (a) promote lactobacillus increase；(b) Bifidobacterium is promoted to increase；(c) promote intestines ball Bacterium is reduced；(d) enterobacteria is promoted to reduce.
- 6. lactobacillus plantarum described in claim 1 is preparing the application in antioxidant.
- 7. application as claimed in claim 6, it is characterised in that: the function of the antioxidant is to remove free radical.
- 8. the use as claimed in claim 7, it is characterised in that: the free radical be hydroxy radical and or DPPH free radical and Or ultra-oxygen anion free radical.
- 9. a kind of antioxidant, active constituent is lactobacillus plantarum described in claim 1.
- 10. antioxidant as claimed in claim 9, it is characterised in that: the function of the antioxidant is to remove free radical.
- 11. antioxidant as claimed in claim 10, it is characterised in that: the free radical be hydroxy radical and or DPPH from By base and or ultra-oxygen anion free radical.
- 12. lactobacillus plantarum described in claim 1 is preparing the application in food, health care product or drug.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510464648.9A CN105062918B (en) | 2015-07-31 | 2015-07-31 | One lactobacillus plantarum and its application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510464648.9A CN105062918B (en) | 2015-07-31 | 2015-07-31 | One lactobacillus plantarum and its application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105062918A CN105062918A (en) | 2015-11-18 |
| CN105062918B true CN105062918B (en) | 2019-01-08 |
Family
ID=54492439
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510464648.9A Active CN105062918B (en) | 2015-07-31 | 2015-07-31 | One lactobacillus plantarum and its application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105062918B (en) |
Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105505814B (en) * | 2015-11-30 | 2020-01-17 | 南昌大学 | Lactobacillus plantarum for delaying senescence |
| CN105969681A (en) * | 2016-03-28 | 2016-09-28 | 四川农业大学 | Highly oxidation-resistant, cholate-tolerant and acid-resistant lactobacillus plantarum JR4 and application thereof |
| CN105920049A (en) * | 2016-06-02 | 2016-09-07 | 扬州大学 | Application of lactobacillus rhamnosus grx19 in regulation of intestinal floras |
| CN106497849B (en) * | 2016-12-19 | 2019-06-04 | 上海理工大学 | A kind of lactobacillus plantarum with high adhesive force |
| CN108165512B (en) * | 2018-02-05 | 2019-12-13 | 富乐顿生物工程科技（北京）有限公司 | Extracellular polysaccharide-producing space lactobacillus plantarum SS18-119 and application thereof in improving biological antioxidant activity |
| CN110760464B (en) * | 2019-11-15 | 2022-05-10 | 湖南农业大学 | Lactobacillus plantarum and application thereof |
| CN113234622A (en) * | 2021-04-30 | 2021-08-10 | 四川高福记生物科技有限公司 | Lactobacillus plantarum 360 with function of regulating intestinal flora and application thereof |
| CN114574408B (en) * | 2022-05-06 | 2022-07-22 | 山东锦鲤生物工程有限公司 | Probiotic SEUNEU-107 and application |
| CN115044514A (en) * | 2022-06-30 | 2022-09-13 | 中国农业科学院饲料研究所 | Lactobacillus plantarum, pet health product and using method thereof |
| CN116162579A (en) * | 2023-04-19 | 2023-05-26 | 东北林业大学 | Preparation method of three probiotic culture media and preparation method of microbial inoculum with DPPH (digital versatile phosphate) scavenging capacity |
Citations (4)
| 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 |
| CN102358888A (en) * | 2011-09-20 | 2012-02-22 | 福建省农业科学院农业工程技术研究所 | Lactobacillus plantarum R23 |
| CN102533618A (en) * | 2012-02-28 | 2012-07-04 | 江南大学 | Lactobacillus plantarum CCFM8724 and application thereof |
| CN102586155A (en) * | 2012-03-07 | 2012-07-18 | 江南大学 | Lactobacillus plantarum N13 and use thereof |
-
2015
- 2015-07-31 CN CN201510464648.9A patent/CN105062918B/en active Active
Patent Citations (4)
| 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 |
| CN102358888A (en) * | 2011-09-20 | 2012-02-22 | 福建省农业科学院农业工程技术研究所 | Lactobacillus plantarum R23 |
| CN102533618A (en) * | 2012-02-28 | 2012-07-04 | 江南大学 | Lactobacillus plantarum CCFM8724 and application thereof |
| CN102586155A (en) * | 2012-03-07 | 2012-07-18 | 江南大学 | Lactobacillus plantarum N13 and use thereof |
Non-Patent Citations (5)
| Title |
|---|
| 两株自选乳酸菌的益生特性;林晓姿,等;《北京工商大学学报（自然科学版）》;20120131;第30卷(第1期);30-35 * |
| 乳酸菌的抗氧化活性和耐酸耐胆盐性能的研究;刘宏宇,等;《食品工业科技》;20141231(第2期);92-96,99 * |
| 植物乳杆菌的黏附特性研究及其在益生菌干酪中的应用;张莉;《中国博士学位论文全文数据库 工程科技I辑》;20130815(第8期);B024-73 * |
| 益生性植物乳杆菌的体外抗氧化活性研究;马爽,等;《中国乳品工业》;20131231;第41卷(第4期);10-14 * |
| 酸菜中降胆固醇功能植物乳杆菌的体外筛选;于志会,等;《食品与生物技术学报》;20110531;第30卷(第3期);398-402 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105062918A (en) | 2015-11-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105062918B (en) | One lactobacillus plantarum and its application | |
| CN101864375B (en) | Lactobacillu plantarurn with function of reducing cholesterol and application thereof | |
| CN101338283B (en) | Lactobacillus casei and applications thereof in solid-state fermentation | |
| CN109536415A (en) | A kind of Lactobacillus rhamnosus and its application | |
| CN103421715B (en) | Lactobacillus rhamnosus and application thereof | |
| CN102242184B (en) | Detection method for viable counts of strains in tetravaccine tablets | |
| CN113080461A (en) | Probiotics with function of inhibiting growth of proteus mirabilis as well as fermentation liquor and application thereof | |
| CN110777097A (en) | Lactobacillus strain with strong acid resistance and screening and fermenting processes thereof | |
| CN111849836B (en) | Lactobacillus rhamnosus with antioxidant function and application thereof | |
| CN115747111A (en) | Pediococcus pentosaceus and application thereof | |
| CN116676225B (en) | Lactobacillus rhamnosus LR-28 strain with nerve soothing and sleep aiding effects, fermentation product, hypnotic fungus group mixture and application | |
| CN114642686B (en) | Composite probiotics and its functions of delaying senility and resisting oxidation | |
| CN103343107A (en) | Human lactobacillus casei gr x 12 with antioxidant function and application thereof | |
| CN109022313A (en) | One lactobacillus plantarum | |
| CN108410761A (en) | One plant has drop nitrite, oxidation resistant Lactobacillus rhamnosus and method for screening and separating | |
| CN105624071A (en) | Lactobacillus salivarius XJP2 and application thereof | |
| CN110106113A (en) | One plant of Lactobacillus kefir MSR101 and its application | |
| CN102108337B (en) | Lactobacillus casei and application thereof | |
| CN113215067B (en) | VBNC (viable but non-viable) state lactobacillus brevis CSHRR5-3 strain and application thereof | |
| CN103667124A (en) | Lactic acid bacteria with capacity of degrading creatinine and urea and screening method for lactic acid bacteria | |
| CN113508907A (en) | Application of heat-resistant lactobacillus fermentum in preparation of defecation promoting food or medicine | |
| CN111154694A (en) | Microbial fermentation inoculant and preparation method and application thereof | |
| CN115838652B (en) | Lactobacillus reuteri LR21 and application thereof | |
| CN106497849B (en) | A kind of lactobacillus plantarum with high adhesive force | |
| CN117305150A (en) | Bifidobacterium animalis BLH1, direct vat set starter and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |