CN105062918A - Lactobacillus plantarum and applications thereof - Google Patents
Lactobacillus plantarum and applications thereof Download PDFInfo
- Publication number
- CN105062918A CN105062918A CN201510464648.9A CN201510464648A CN105062918A CN 105062918 A CN105062918 A CN 105062918A CN 201510464648 A CN201510464648 A CN 201510464648A CN 105062918 A CN105062918 A CN 105062918A
- Authority
- CN
- China
- Prior art keywords
- plant lactobacillus
- application
- lactobacillus
- product
- bacterial strain
- 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.)
- Granted
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
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses a lactobacillus plantarum and applications thereof, and specifically provides the lactobacillus plantarum NC 301 with the preservation register number CGMCC NO.11013. The application of the lactobacillus plantarum NC 301 in preparing a product for regulating the intestinal flora is also within the scope of protection of the invention. The invention further provides the application of the lactobacillus plantarum NC 301 in preparing an antioxidant. The bacterial strain provided by the invention is a good novel lactobacillus strain, and also can be taken as a probiotic with functions of regulating the intestinal flora and improving the intestinal functions to be applied to the fields of food, health products and medicines, and therefore, the application prospect is very broad.
Description
Technical field
The invention belongs to microbial technology field, be specifically related to a lactobacillus plantarum and application thereof.
Background technology
Chinese tradition leavened food is with a long history, unique flavor, enjoys the favor of human consumer.Traditional fermented food comprises fermented vegetables, fermented meat prods, cultured milk prod, white wine and fermented seasonings etc., is the important component part of China's foodstuffs industry.Milk-acid bacteria in traditional fermented food is not only conducive to local flavor, the mouthfeel and quality etc. of improving leavened food, and some milk-acid bacterias also have excellent probiotic properties.
The superior geographical environment in Changbaishan area creates local abundant natural resources, defines the leavened food having characteristic, as pickles, salted vegetables etc.Naturally tame through long-term, milk-acid bacteria becomes the dominant microflora in these traditional cuisines, is the good source of probiotic bacterium.
Summary of the invention
The object of this invention is to provide a lactobacillus plantarum and application thereof.
Plant lactobacillus provided by the invention (Lactobacillusplantarum) NC301, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on 06 23rd, 2015 and (be called for short CGMCC, address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica), preservation registration number is CGMCCNO.11013.Plant lactobacillus (Lactobacillusplantarum) NC301CGMCCNO.11013, is called for short plant lactobacillus NC301.
The application of plant lactobacillus NC301 in the product for the preparation of regulating intestinal canal flora also belongs to protection scope of the present invention.The function of described " product for regulating intestinal canal flora " for (a) and or (b) and or (c) and or (d): (a) promotes that Bacterium lacticum increases; B () promotes that bifidus bacillus increases; C () promotes that faecalis reduces; D () promotes that enterobacteria reduces.
The present invention also protects a kind of product for regulating intestinal canal flora, and its activeconstituents is plant lactobacillus NC301.The function of described " product for regulating intestinal canal flora " for (a) and or (b) and or (c) and or (d): (a) promotes that Bacterium lacticum increases; B () promotes that bifidus bacillus increases; C () promotes that faecalis reduces; D () promotes that enterobacteria reduces.
The present invention goes back protective plant Bacterium lacticum NC301 and is preparing the application in antioxidant.The function of described antioxidant is scavenging free radicals.Described free radical be hydroxy radical qiao and or DPPH free radical and or ultra-oxygen anion free radical.
The present invention also protects a kind of antioxidant, and its activeconstituents is plant lactobacillus NC301.The function of described antioxidant is scavenging free radicals.Described free radical be hydroxy radical qiao and or DPPH free radical and or ultra-oxygen anion free radical.
The present invention goes back protective plant Bacterium lacticum NC301 and is preparing the application in food, healthcare products or medicine.Described food specifically can be Yoghourt.
The present invention is separated and obtains a lactobacillus plantarum from traditional zymotic Tian jin cabbage pickled in sweet and sour, by external acid tolerance and cholate tolerance, Adhering capacity, antioxygenation and improve the test such as intestinal function and regulating intestinal canal flora, confirm that this bacterial strain is the Lactic Acid Bacteria new strains that a strain has potential probiotic properties, can be applied to food, healthcare products and medicine field as regulating intestinal canal flora, the probiotic bacterium of improving intestinal function, application prospect is very wide.
Accompanying drawing explanation
Fig. 1 is plant lactobacillus NC301 photo under an optical microscope.
Fig. 2 is the photo of plant lactobacillus NC301 under transmission electron microscope.
Fig. 3 is the regulating power of plant lactobacillus NC301 to enteron aisle flora.
Embodiment
Following embodiment is convenient to understand the present invention better, but does not limit the present invention.Experimental technique in following embodiment, if no special instructions, is ordinary method.Test materials used in following embodiment, if no special instructions, is and purchases available from routine biochemistry reagent shop.Quantitative test in following examples, all arranges and repeats experiment for three times, results averaged.Bilein: the Henan big biological medicine company limited-liability company of profit, production code member is P333004, and No. CAS is 8008-63-7.Caco-2 cell: the American Type Culture Collection council of Chinese Academy of Sciences cell bank.
BCP liquid nutrient medium: yeast extract paste 2.5g/L, peptone 5g/L, glucose 5g/L, purpurum bromocresolis 0.04g/L, solvent is water, pH7.0; 121 DEG C of sterilizing 15min.The basis of BCP liquid nutrient medium is added agar to 15g/L, be BCP solid medium.
MRS liquid nutrient medium: peptone 10g/L, extractum carnis 10g/L, yeast extract paste 5g/L, KH
2pO
42g/L, sodium acetate 5g/L, Trisodium Citrate 5g/L, MgSO
47H
2o0.2g/L, MnSO
44H
2o0.05g/L, tween 80 1mL, glucose 20g/L, solvent is water, pH6.6; 121 DEG C of sterilizing 15min.The basis of MRS liquid nutrient medium is added agar to 15g/L, be MRS solid medium
The preparation method of the PBS damping fluid of pH7.2: NaCl8g, KCl0.2g, Na
2hPO
41.42g, K
2hPO
40.27g, adds water and is settled to 1L, and concentrated hydrochloric acid adjusts pH7.2.
The preparation method of the PBS damping fluid of pH7.4: NaCl8g, KCl0.2g, Na
2hPO
41.42g, K
2hPO
40.27g, adds water and is settled to 1L, and concentrated hydrochloric acid adjusts pH7.4.
Lactobacillus rhamnosus LGG, is the existing probiotic bacterium of a strain, represents with LGG.Mention the document of lactobacillus rhamnosus LGG: Zhang Qilin, Ma Chunli, Li Aili, Cao Nan, Song Lanlan, the application of lactobacillus rhamnosus LGG in Cottage cheese, " foodstuffs industry " 2015 years 04 phases.
The separation andpreconcentration of embodiment 1, bacterial strain
Sample for strains separation is the traditional zymotic Tian jin cabbage pickled in sweet and sour of Yanbian of China.
Repeatedly streak culture through BCP solid medium flat board, single colony inoculation that picking produces yellow circle continues to cultivate on MRS solid medium flat board, through MRS solid medium flat board streak culture purifying repeatedly, obtains the bacterial strain of many strains pure culture.A wherein strain equal called after NC301 bacterial strain.
Two, the qualification of bacterial strain
Fig. 1 is shown in by NC301 bacterial strain photo under an optical microscope.Somatic cells is nose circle direct rod shape, and great majority are single, paired or are cell chains.
Fig. 2 is shown in by the photo of NC301 bacterial strain under transmission electron microscope.Elongated rod shape, cell walls, cytolemma are complete, and kytoplasm is even.
NC301 bacterial strain is the bacillus do not moved.
NC301 bacterial strain optimum growth temperature 37 ~ 42 DEG C, appropriate pH is 5.0 ~ 7.0.
The physiological and biochemical property of NC301 bacterial strain: Gram-positive, negative catalase, can grow 15 DEG C and 45 DEG C, tolerance 6.5%NaCl, not hydrolyzed starch, not liquefy gelatin, do not produce hydrogen sulfide, glucose fermentation produces acid not aerogenesis, and benzidine test is negative, indole test is negative, and voges-Proskauer test is positive.
API50CH (French Mei Liai company) qualification result shows that NC301 bacterial strain is plant lactobacillus.NC301 bacterial strain can utilize: ribose, semi-lactosi, glucose, fructose, seminose, N.F,USP MANNITOL, sorbyl alcohol, amygdaloside, arbutin, N-acetyl-glucosamine, seven leaf-alcohols, salicin, cellobiose, maltose, lactose, melibiose, sucrose, trehalose, melizitose, raffinose, geraniol, D-R alcohol, gluconate.NC301 bacterial strain can not utilize: pectinose, wood sugar, ribitol, sorbose, rhamnosyl, galactitol, inositol, Alpha-Methyl-D-Glucose glycosides, Alpha-Methyl-D-MANNOSE glycosides, synanthrin, starch, glycogen, Xylitol, D-turanose, D-Tag, D-rice threose, rock sugar, L-arabinose alcohol, 2-keto-D-gluconate salt, 5-keto-D-gluconate salt.
The 16srDNA partial sequence of NC301 bacterial strain is as shown in sequence in sequence table 1.
Above qualification result shows, NC301 bacterial strain belongs to plant lactobacillus (Lactobacillusplantarum).
Three, the preservation of bacterial strain
Plant lactobacillus provided by the invention (Lactobacillusplantarum) NC301, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on 06 23rd, 2015 and (be called for short CGMCC, address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica), preservation registration number is CGMCCNO.11013.Plant lactobacillus (Lactobacillusplantarum) NC301CGMCCNO.11013, is called for short plant lactobacillus NC301, represents with NC301.
The tolerance of embodiment 2, plant lactobacillus NC301 measures
1, plant lactobacillus NC301 is seeded to MRS liquid nutrient medium, obtains the bacteria suspension (bacteria concentration in bacteria suspension: log
10cfu/ml=8.57 ± 0.17), bacteria suspension is divided into 4 groups (often organizing 5 re-treatments), proceeds as follows respectively:
First group: adjust pH to be 2.0,37 DEG C of quiescent culture 3h with hydrochloric acid;
Second group: adjust pH to be 3.0,37 DEG C of quiescent culture 3h with hydrochloric acid;
3rd group: add bilein and make its concentration be 0.3g/100ml, 37 DEG C of quiescent culture 3h;
4th group: add bilein and make its concentration be 0.6g/100ml, 37 DEG C of quiescent culture 4h;
After completing aforesaid operations, get 50 μ l and be coated with MRS solid medium flat board, enumeration after 37 DEG C of quiescent culture 48h.
The results are shown in Table 1.Plant lactobacillus NC301 has certain tolerance to pH2.0, survival rate 20.63%.Plant lactobacillus NC301 has good tolerance effect to pH3.0, and survival rate is still more than 80%.Plant lactobacillus NC301 all has stronger tolerance to the bovine bile of 0.3g/100ml and 0.6g/100ml.
2, lactobacillus rhamnosus LGG is seeded to MRS liquid nutrient medium, obtains the bacteria suspension (bacteria concentration in bacteria suspension: log
10cfu/ml=8.25 ± 0.37), bacteria suspension is divided into 4 groups (often organizing 5 re-treatments), proceeds as follows respectively:
First group: adjust pH to be 2.0,37 DEG C of quiescent culture 3h with hydrochloric acid;
Second group: adjust pH to be 3.0,37 DEG C of quiescent culture 3h with hydrochloric acid;
3rd group: add bilein and make its concentration be 0.3g/100ml, 37 DEG C of quiescent culture 3h;
4th group: add bilein and make its concentration be 0.6g/100ml, 37 DEG C of quiescent culture 4h;
After completing aforesaid operations, get 50 μ l and be coated with MRS solid medium flat board, enumeration after 37 DEG C of quiescent culture 48h.
The results are shown in Table 1.
Table 1 bacterial strain is to tolerance that is sour and cholate
The Adhering capacity of embodiment 3, plant lactobacillus NC301
Surface hydrophobicity is relevant with the Adhering capacity of bacterial strain.The hydrophobic property on bacterial strain surface is one of important factor affecting bacterial strain intestinal epithelial cell Adhering capacity.
One, hydrophobicity measures
1, get test strains, with the PBS damping fluid suspension thalline of pH7.2, obtain OD
600nmthe bacteria suspension of=0.4.
2, get the bacteria suspension that 3mL step 1 obtains, mix with 1mL organic reagent, whirlpool concussion 30s, then room temperature leaves standstill 30min, water intaking phase.
3, get the aqueous phase that step 2 obtains, the absorbance measured under 600nm (is designated as A
x).
The hydrophobic ability of bacterial strain (%)=[1-(A
x/ A
0)] × 100%; A
0=0.4.
Test strains is plant lactobacillus NC301 or lactobacillus rhamnosus LGG.
Organic reagent is dimethylbenzene, trichloromethane or ethyl acetate.
Carry out three revision tests, results averaged.
The results are shown in Table 2.Compared with lactobacillus rhamnosus LGG, plant lactobacillus NC301 p-Xylol, chloroform and ethyl acetate all have stronger hydrophobic ability, show that its surface has extremely strong electron donor characteristic.
Two, Caco-2 cell adhesion test
Test strains is plant lactobacillus NC301 or lactobacillus rhamnosus LGG.
1, in 24 porocyte culture plates, the DMEM nutrient solution containing 10% (volume ratio) new-born calf serum, 100U/mL penicillin and 100U/mL Streptomycin sulphate is adopted to cultivate Caco-2 cell, grow to monolayer cell adherent time count, every porocyte number is 3.5 × 10
5individual.
2, get test strains, with the PBS damping fluid suspension thalline of pH7.2, obtaining bacteria concentration is 3.4 × 10
9the bacteria suspension of cfu/m1.
3, packet transaction
Test group: get the 24 porocyte culture plates that step 1 obtains, every hole adds bacteria suspension that 0.5mL step 2 obtains and the fresh DMEM nutrient solution of 0.5mL, puts 37 DEG C, 5%CO
2, 90% humidity environment in stationary incubation 2h, then use the PBS damping fluid rinsing cell 5 times (to remove the bacterium of not sticking) of pH7.4.
Control group: the bacteria suspension replacing step 2 to obtain with the PBS damping fluid of equal-volume pH7.2, other same test group.
Often group arranges three re-treatments.
4, after completing steps 3, get described 24 orifice plates, fix 30min with 0.4% paraformaldehyde, gramstaining, microscopy.
5, after completing steps 4, get described 24 orifice plates, every hole adds 0.2mL0.25% trysinization 10min, then every hole adds the DMEM nutrient solution of 0.125mL containing 20% (volume ratio) serum with termination reaction, then every hole adds 0.175mL0.1% (volume ratio) the TritonX-100 aqueous solution, room temperature leaves standstill 10min (with lysing cell), then carries out plate count.
Bacterial count × 100% added in sum/every hole of bacterium is attached in adherence rate (%)=every hole.
The results are shown in Table 2.Compared with lactobacillus rhamnosus LGG, plant lactobacillus NC301 Adhering capacity is comparatively strong, shows that plant lactobacillus NC301 has stronger Adhering capacity to intestinal epithelial cells.
Table 2 surface hydrophobicity and Caco-2 cell adhesion ability
Embodiment 4, plant lactobacillus NC301 are to the regulating effect of intestinal microflora
Intestinal microflora regulating power is the important of probiotic bacterium and basic function.The realization of probiotic bacterium function except bacterial strain self have functional except, mainly by regulating the flora of host intestine, intestinal microflora is often compared to the organ of human body, there is the synthesis and metabolism that promote nutritive element, activate and immunity moderation system, alleviate enteron aisle and do not accommodate diarrhoea, the functions such as the generation of prevention and minimizing intestinal tract infections and body self-regeneration.
Kunming mice (4 week age, male, 18-22g), purchased from high-new medical faunae center, Changchun.
Kunming mice is divided at random 2 groups (often organizing 10), is handled as follows respectively:
Test group: (volume is 0.4mL to gavage every day plant lactobacillus bacterium liquid, containing 4 × 10
8cfu plant lactobacillus NC301, solvent is the PBS damping fluid of pH7.2), continuous gavage 14 days (testing the 1 to 14 day), then normally raises;
Control group: the PBS damping fluid (volume is 0.4mL) of a gavage every day pH7.2, continuous gavage 14 days (testing the 1 to 14 day), then normally raises.
Respectively at test the 0th day, within the 7th day, the 14th day, the 16th day, the 21st day, gather the ight soil of mouse, detect the quantity of wherein Bacterium lacticum, bifidus bacillus, faecalis and enterobacteria.
Detect Bacterium lacticum and adopt the mode selecting slat chain conveyor to be combined with morphologic observation.The selection flat board adopted is Bacterium lacticum nutrient agar (LBS), and preparation method is as follows: pancreas casein peptone 10g, yeast leaching powder 5g, potassium primary phosphate 6g, ferrous sulfate 0.034g, magnesium sulfate 0.575g, glucose 20g, sodium acetate 25g, ammonium citrate 2g, manganous sulfate 0.12g, tween 80 1mL, agar 12g, hydrochloric acid adjust pH to 5.5, heated and stirred is dissolved in 1000mL distilled water, 115 DEG C of autoclaving 15min.Culture condition: 37 DEG C, 48h.The morphological specificity of Bacterium lacticum: white or oyster white, the sporeless bacterium that gramstaining is positive.
Detect bifidus bacillus and adopt the mode selecting slat chain conveyor to be combined with morphologic observation.The selection flat board adopted is Medium of Bifidobacterium (MRS+NNLP), is the MRS solid medium containing Nalidixic Acid 15mg/L, neomycinsulphate 100mg/L, paromomycin sulfate 200mg/L, lithium chloride 3g/L, L-cysteine hydrochloride 500mg/L.Culture condition: 37 DEG C of anaerobism, 48h.The morphological specificity of bifidus bacillus: micro white, smooth surface, protruding, the forked or bar-shaped bacterium colony that gramstaining is positive.
Detect enterobacteria and adopt the mode selecting slat chain conveyor to be combined with morphologic observation.The selection flat board adopted is violet red bile dextrose agar substratum (VioletRedBileDextroseAgar, VRBDA), and preparation method is as follows: yeast powder 3g, peptone 7g, sodium-chlor 5g, glucose 10g, cholate 1.5g, Viola crystallina 0.002g, toluylene red 0.03g, agar 12g, hydrochloric acid adjust pH 7.3, heating for dissolving, in 1000mL distilled water, is boiled and is not exceeded 2min, when being chilled to about 50 DEG C, impouring sterilized petri dishes.Culture condition: 37 DEG C, 24h.The morphological specificity of enterobacteria: ferment lactose, the bacterium colony that gramstaining is negative.
Detect faecalis and adopt the mode selecting slat chain conveyor to be combined with morphologic observation.The selection flat board adopted is enterococcosel agar (cholate-Vitamin C2-sodium azide agar) substratum (BileEsculinAzideAgar, BEA), and preparation method is as follows: beef extract powder 3g, pancreas casein peptone 17g, yeast powder 5g, bilein 10g, sodium-chlor 5g, Vitamin C2 1g, ferric ammonium citrate 0.5g, sodiumazide 0.25g, Trisodium Citrate 1g, agar 13g, hydrochloric acid adjust pH 7.1, heating for dissolving in 1000mL distilled water, 121 DEG C of autoclaving 15min.Culture condition: 37 DEG C, 48h.Enterococcal morphological specificity: obviously brown circle, the bacterium colony that gramstaining is positive.
(filled black post represents control group, and grey packed column represents test group to the results are shown in Figure 3; A: Bacterium lacticum; B: faecalis; C: bifidus bacillus; D: enterobacteria).Compared with control group, in the ight soil of test group mouse, the quantity of Bacterium lacticum and bifidus bacillus obviously increases, and the quantity of faecalis and enterobacteria reduces.Result shows, plant lactobacillus NC301 can promote that in mouse intestinal, Bacterium lacticum and bifidus bacillus are increased, and faecalis and enterobacteria reduce.
The antioxidant activity in vitro of embodiment 5, plant lactobacillus NC301
One, Hydroxyl radical-scavenging ability measures (Fenton method)
1, to suspend bacterium to be measured (bacterium to be measured is plant lactobacillus NC301 or lactobacillus rhamnosus LGG) with the PBS damping fluid of pH7.2, obtaining bacteria concentration is 1 × 10
10the bacteria suspension of cfu/mL.
2, packet transaction
Test group: get test tube, adds the 0.435mM BG aqueous solution 1mL, 0.5mM ferrous sulfate aqueous solution 2mL, 3% (volume ratio) aqueous hydrogen peroxide solution 1.5mL (Fe successively
2+the oxidation system formed with hydrogen peroxide is called Fenton reagent) and the bacteria suspension prepared of 1mL step 1, mix, then 37 DEG C of waters bath with thermostatic control insulation 20min, then measure the absorbancys of supernatant liquor at 624nm place;
Control group first: the bacteria suspension replacing step 1 to prepare with the PBS damping fluid of equal-volume pH7.2, other same test group;
Control group second: the bacteria suspension replacing step 1 to prepare with the PBS damping fluid of equal-volume pH7.2, replaces the BG aqueous solution with equal-volume water, replaces aqueous hydrogen peroxide solution, other same test group with equal-volume water;
Often group arranges five re-treatments.
Bacterial strain is to Scavenging activity (%)=[(As-Ao)/(A-Ao)] × 100% of hydroxy radical qiao;
In formula: the absorbance of As test group; Ao represents the absorbance of control group first; A represents the absorbance of control group second.
Two, DPPH radical scavenging activity measures
1, to suspend bacterium to be measured (bacterium to be measured is plant lactobacillus NC301 or lactobacillus rhamnosus LGG) with the PBS damping fluid of pH7.2, obtaining bacteria concentration is 1 × 10
10the bacteria suspension of cfu/mL.
2, packet transaction
Test group: (full name of DPPH is 1 to get 2mL0.5mMDPPH solution, 1-phenylbenzene-2-trinitrophenyl-hydrazine, the solvent of DPPH solution is methyl alcohol), add the bacteria suspension that 1mL step 1 obtains, mix, then room temperature lucifuge places 30min, then 4 DEG C, the centrifugal 10min of 8000g, get supernatant liquor, measure the absorbancy of supernatant liquor at 517nm place;
Negative control group: the bacteria suspension replacing step 1 to prepare with the PBS damping fluid of equal-volume pH7.2, other same test group;
Blank group: replace DPPH solution with equal-volume methyl alcohol, other same test group;
Often group arranges five re-treatments.
Bacterial strain is to Scavenging activity (%)=[1-(A of DPPH free radical
sample-A
blank)/A
contrast] × 100%;
In formula: A
samplerepresent the absorbance of test group; A
blankrepresent the absorbance of blank group; A
contrastrepresent the absorbance of negative control group.
Three, superoxide anion Scavenging activity measures
1, to suspend bacterium to be measured (bacterium to be measured is plant lactobacillus NC301 or lactobacillus rhamnosus LGG) with the PBS damping fluid of pH7.2, obtaining bacteria concentration is 1 × 10
10the bacteria suspension of cfu/mL.
2, packet transaction
Test group: get test tube, add bacteria suspension, the 2.8mL3mM diethylenetriamine pentaacetic acid aqueous solution, the 0.2mL1.2mM pyrogallol aqueous solution that 0.3mL step 1 obtains successively, mix, then 25 DEG C of water bath with thermostatic control insulation 10min, then measure the absorbancy at 325nm place;
Control group first: replace the pyrogallol aqueous solution with equal-volume water, other same test group;
Control group second: the bacteria suspension replacing step 1 to prepare with the PBS damping fluid of equal-volume pH7.2, other same test group;
Control group third: the bacteria suspension replacing step 1 to prepare with the PBS damping fluid of equal-volume pH7.2, replaces the pyrogallol aqueous solution with equal-volume water, other same test group;
Often group arranges five re-treatments.
Bacterial strain is to Scavenging activity (%)=[1-(A of superoxide anion
11-A
10)/(A
01-A
00)] × 100%
In formula: A
11the absorbance of test group; A
10represent the OD value of control group first; A
01represent the OD value of control group second; A
00represent the OD value of control group third.
Step one, step 2 and step 3 the results are shown in Table 3.The clearance rate of plant lactobacillus NC301 to hydroxy radical qiao, DPPH free radical, ultra-oxygen anion free radical has all exceeded 50.0%.
Table 3 radical scavenging activity
Bacterial strain | Hydroxy radical qiao (%) | 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 (9)
1. plant lactobacillus (Lactobacillusplantarum) NC301, its deposit number is CGMCCNO.11013.
2. the application of plant lactobacillus according to claim 1 in the product for the preparation of regulating intestinal canal flora.
3., for a product for regulating intestinal canal flora, its activeconstituents is plant lactobacillus according to claim 1.
4. application as claimed in claim 2 or product as claimed in claim 3, is characterized in that: the function of described " product for regulating intestinal canal flora " for (a) and or (b) and or (c) and or (d): (a) promotes that Bacterium lacticum increases; B () promotes that bifidus bacillus increases; C () promotes that faecalis reduces; D () promotes that enterobacteria reduces.
5. plant lactobacillus according to claim 1 is preparing the application in antioxidant.
6. an antioxidant, its activeconstituents is plant lactobacillus according to claim 1.
7. application as claimed in claim 5 or antioxidant as claimed in claim 6, is characterized in that: the function of described antioxidant is scavenging free radicals.
8. application as claimed in claim 7 or product, is characterized in that: described free radical be hydroxy radical qiao and or DPPH free radical and or ultra-oxygen anion free radical.
9. plant lactobacillus according to claim 1 is preparing the application in food, healthcare products or medicine.
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 true CN105062918A (en) | 2015-11-18 |
CN105062918B 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) |
Cited By (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105505814A (en) * | 2015-11-30 | 2016-04-20 | 南昌大学 | Lactobacillus plantarum capable of delaying aging |
CN105920049A (en) * | 2016-06-02 | 2016-09-07 | 扬州大学 | Application of lactobacillus rhamnosus grx19 in regulation of intestinal floras |
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 |
CN106497849A (en) * | 2016-12-19 | 2017-03-15 | 上海理工大学 | A kind of Lactobacillus plantarum with high adhesive force |
CN108165512A (en) * | 2018-02-05 | 2018-06-15 | 富乐顿生物工程科技(北京)有限公司 | A kind of extracellular polysaccharide spatial plant lactobacillus SS18-119 and its application in biological antioxidant activity is improved |
CN110760464A (en) * | 2019-11-15 | 2020-02-07 | 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 |
CN114574408A (en) * | 2022-05-06 | 2022-06-03 | 山东锦鲤生物工程有限公司 | Probiotic SEUNEU-107 and application thereof |
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 |
---|
于志会,等: "酸菜中降胆固醇功能植物乳杆菌的体外筛选", 《食品与生物技术学报》 * |
刘宏宇,等: "乳酸菌的抗氧化活性和耐酸耐胆盐性能的研究", 《食品工业科技》 * |
张莉: "植物乳杆菌的黏附特性研究及其在益生菌干酪中的应用", 《中国博士学位论文全文数据库 工程科技I辑》 * |
林晓姿,等: "两株自选乳酸菌的益生特性", 《北京工商大学学报(自然科学版)》 * |
马爽,等: "益生性植物乳杆菌的体外抗氧化活性研究", 《中国乳品工业》 * |
Cited By (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105505814A (en) * | 2015-11-30 | 2016-04-20 | 南昌大学 | Lactobacillus plantarum capable of delaying aging |
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 |
CN106497849A (en) * | 2016-12-19 | 2017-03-15 | 上海理工大学 | A kind of Lactobacillus plantarum with high adhesive force |
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 |
CN108165512A (en) * | 2018-02-05 | 2018-06-15 | 富乐顿生物工程科技(北京)有限公司 | A kind of extracellular polysaccharide spatial plant lactobacillus SS18-119 and its application in biological antioxidant activity is improved |
CN110760464A (en) * | 2019-11-15 | 2020-02-07 | 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 |
CN114574408A (en) * | 2022-05-06 | 2022-06-03 | 山东锦鲤生物工程有限公司 | Probiotic SEUNEU-107 and application thereof |
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 |
Also Published As
Publication number | Publication date |
---|---|
CN105062918B (en) | 2019-01-08 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN105062918A (en) | Lactobacillus plantarum and applications thereof | |
CN103013879B (en) | Bacterial strain having capacity of giving high yield of Gamma-aminobutyric acid | |
CN101338283B (en) | Lactobacillus casei and applications thereof in solid-state fermentation | |
CN101864375B (en) | Lactobacillu plantarurn with function of reducing cholesterol and application thereof | |
CN101144065B (en) | Oxidation resistant Lactobacillus casei capable of resisting hydrogen peroxide and eliminating free radical, and use thereof | |
CN104651268B (en) | A kind of Lactobacillus plantarum and its application | |
CN103421715B (en) | Lactobacillus rhamnosus and application thereof | |
CN105132322A (en) | Lactobacillus plantarum and application thereof | |
CN102242184B (en) | Detection method for viable counts of strains in tetravaccine tablets | |
CN103114051A (en) | Roy bacterium lacticum and application thereof | |
CN105533544A (en) | Natural fruit and vegetable enzyme product | |
CN105255752A (en) | Lactobacillus plantarum ZDY2013 with high acid resistance | |
CN107937305B (en) | Butyric acid-resistant strong-stress-resistant clostridium butyricum microbial inoculum and application thereof | |
CN105400727A (en) | Lactobacillus paracasei with antioxidant activity and application thereof | |
CN104928208A (en) | Lactobacillus plantarum Lp90, and screening method and application thereof | |
CN106957810A (en) | A kind of Pediococcus acidilactici and application thereof | |
CN103540545A (en) | Pediococcus pentosaceus and application thereof | |
CN110106119A (en) | The Lactobacillus rhamnosus M9 of one plant of isolated from mother's milk and its application | |
CN100368530C (en) | Bifidobacteria exocellular polysaccharide and its production method and special purpose production strain | |
CN102021127A (en) | Lactobacillus paracasei 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 | |
CN103343107A (en) | Human lactobacillus casei gr x 12 with antioxidant function and application thereof | |
CN105838658B (en) | A method of improving lactic acid bacteria biomass under high salt conditions | |
CN107058171A (en) | Lactobacillus pentosus and application with reduction cholesterol reduction triglycerides effect | |
CN108893416A (en) | It is a kind of degrade vomitoxin saccharomycete and its application |
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 |