CN105567601A - Lactobacillus plantarum and application thereof - Google Patents
Lactobacillus plantarum and application thereof Download PDFInfo
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- CN105567601A CN105567601A CN201610066292.8A CN201610066292A CN105567601A CN 105567601 A CN105567601 A CN 105567601A CN 201610066292 A CN201610066292 A CN 201610066292A CN 105567601 A CN105567601 A CN 105567601A
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- CN
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- plant lactobacillus
- cholesterol
- lactobacillus plantarum
- application
- bacterium
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- Pending
Links
- 240000006024 Lactobacillus plantarum Species 0.000 title abstract 4
- 235000013965 Lactobacillus plantarum Nutrition 0.000 title abstract 4
- 229940072205 lactobacillus plantarum Drugs 0.000 title abstract 4
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 claims abstract description 42
- 235000012000 cholesterol Nutrition 0.000 claims abstract description 20
- 230000015556 catabolic process Effects 0.000 claims abstract description 12
- 238000006731 degradation reaction Methods 0.000 claims abstract description 12
- 238000002360 preparation method Methods 0.000 claims abstract description 8
- 239000003814 drug Substances 0.000 claims abstract description 3
- 241000186660 Lactobacillus Species 0.000 claims description 27
- 229940039696 lactobacillus Drugs 0.000 claims description 26
- 244000005700 microbiome Species 0.000 claims description 6
- 238000004321 preservation Methods 0.000 abstract description 4
- 230000000593 degrading effect Effects 0.000 abstract 1
- 229940079593 drug Drugs 0.000 abstract 1
- 241000196324 Embryophyta Species 0.000 description 23
- 241000894006 Bacteria Species 0.000 description 19
- ZWLUXSQADUDCSB-UHFFFAOYSA-N phthalaldehyde Chemical compound O=CC1=CC=CC=C1C=O ZWLUXSQADUDCSB-UHFFFAOYSA-N 0.000 description 7
- 230000001580 bacterial effect Effects 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 239000012530 fluid Substances 0.000 description 5
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 4
- 238000009360 aquaculture Methods 0.000 description 4
- 244000144974 aquaculture Species 0.000 description 4
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 229940054441 o-phthalaldehyde Drugs 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 230000031700 light absorption Effects 0.000 description 3
- 235000015097 nutrients Nutrition 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 2
- 241000726221 Gemma Species 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 238000003794 Gram staining Methods 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- 229940040526 anhydrous sodium acetate Drugs 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- YXVFQADLFFNVDS-UHFFFAOYSA-N diammonium citrate Chemical compound [NH4+].[NH4+].[O-]C(=O)CC(O)(C(=O)O)CC([O-])=O YXVFQADLFFNVDS-UHFFFAOYSA-N 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 210000003495 flagella Anatomy 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 235000014655 lactic acid Nutrition 0.000 description 2
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 2
- 235000019341 magnesium sulphate Nutrition 0.000 description 2
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 235000013336 milk Nutrition 0.000 description 2
- 239000008267 milk Substances 0.000 description 2
- 210000004080 milk Anatomy 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- -1 pplication Species 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 108010007979 Glycocholic Acid Proteins 0.000 description 1
- 201000010538 Lactose Intolerance Diseases 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 208000035199 Tetraploidy Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 229960000583 acetic acid Drugs 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
- 230000000172 allergic effect Effects 0.000 description 1
- 208000010668 atopic eczema Diseases 0.000 description 1
- 230000003796 beauty Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 229960000935 dehydrated alcohol Drugs 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 229960004756 ethanol Drugs 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 239000012982 microporous membrane Substances 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 238000012797 qualification Methods 0.000 description 1
- 230000027272 reproductive process Effects 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 238000007127 saponification reaction Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000004460 silage Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- OABYVIYXWMZFFJ-ZUHYDKSRSA-M sodium glycocholate Chemical compound [Na+].C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(=O)NCC([O-])=O)C)[C@@]2(C)[C@@H](O)C1 OABYVIYXWMZFFJ-ZUHYDKSRSA-M 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 238000003805 vibration mixing Methods 0.000 description 1
Classifications
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K2035/11—Medicinal preparations comprising living procariotic cells
- A61K2035/115—Probiotics
Abstract
The invention discloses lactobacillus plantarum. The preservation number of the lactobacillus plantarum is CGMCC NO. 11875. The invention further provides application of the lactobacillus plantarum to degradation of cholesterol and preparation of drugs or health-care products for degrading cholesterol.
Description
Technical field
The invention belongs to microorganism field, is the effect about a kind of newfound plant lactobacillus and cholesterol degradation function thereof.
Background technology
The life of plant lactobacillus and the mankind, animal is in close relations, it is a kind of milk-acid bacteria be common in cream, meat and many vegetable fermentation goods, silage, by stomach and be colonizated in enteron aisle play beneficial effect, such as maintain the balance of intestinal flora, promote absorption of nutrient ingredients, alleviate the several functions such as the formation of lactose intolerance and inhibition tumor cell, the more Application Areas of research report comprises food, feed, health care, makeup, beauty treatment, aquaculture, industrial and agricultural production, biological preservative, probiotics preparation etc. at present.
The newfound plant lactobacillus of the present invention is the Bacterium lacticum extracted in the animal and plant body of aquaculture water, be a kind of plant lactobacillus through qualification, optimum growth temperature is 30 ~ 35, and bacterial classification is straight or curved shaft-like or become chain, optimal pH about 6.5, belongs to homofermentative lactic bacteria.This bacterium and the difference of other milk-acid bacterias are that the viable count of this bacterium is higher, and product acid that can be a large amount of, makes the pH value in water stablize and do not raise; Because this bacterium is anaerobic bacterium (facultative aerobic), the distinctive lactobacillin of energy output in reproductive process, current plant lactobacillus is mainly used at present and changes all types of allergic symptom, improve physique, the medical aspect such as toxin expelling, when studying newfound plant lactobacillus effect, find that it has good effect to cholesterol degradation.
Summary of the invention
Present invention finds a kind of new plant lactobacillus, and find its new purposes.
Particularly, the present invention's extraction and isolation in the animal and plant body of aquaculture water obtains a kind of Bacterium lacticum, through being accredited as a kind of new plant lactobacillus, it is stored in China Committee for Culture Collection of Microorganisms's common micro-organisms center in December, 2015, preservation address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica, and preserving number is CGMCCNO:11875.
Shown in plant lactobacillus 16SrDNA sequence SEQNo1 of the present invention.
What cultivation Bacterium lacticum was used is MRS substratum, and culture medium prescription is: glucose 30g, peptone 10g, yeast powder 7.5g, anhydrous sodium acetate 5g, K2HPO42g, magnesium sulfate 0.8g, manganous sulfate 0.2g, dibasic ammonium citrate 2g, PH7.0-7.5, and culture temperature is 37 DEG C.
The form of bacterial classification
Plant lactobacillus cultivates 48h on substratum, generation bacterium colony figure as shown in Figure 1, colony edge is neat, smooth surface, in milk yellow; By single colonies sampling under an electron microscope (magnification is 1800 times) observe, as shown in Figure 2, cellular form is shaft-like to observations, and gramstaining is accredited as Gram-positive, without gemma and flagellum.
The present invention has found this plant lactobacillus further and it is to the Degradation of cholesterol.
Further, the present invention proposes above-mentioned plant lactobacillus for the preparation of the application on the medicine of cholesterol degradation or healthcare products.
Accompanying drawing explanation
Fig. 1 is plant lactobacillus bacterium colony figure;
Fig. 2 is plant lactobacillus microscopy figure.
Embodiment
Below describe newfound plant lactobacillus in detail by specific experiment and it is to the Degradation of cholesterol.
The present invention's extraction and isolation in the animal and plant body of aquaculture water obtains a kind of Bacterium lacticum, through being accredited as a kind of new plant lactobacillus, it is stored in China Committee for Culture Collection of Microorganisms's common micro-organisms center in December, 2015, preservation address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica, and preserving number is CGMCCNO:11875.
Plant lactobacillus 16SrDNA sequence of the present invention is as shown in sequence table.Plant lactobacillus 16SrDNA sequence of the present invention is as shown in sequence table.By carrying out pcr amplification to the 16SrDNA of this bacterial strain, reclaim amplified production order-checking, the sequence logged in sequence and GenBank database carries out tetraploid rice, and 16SrDNA sequence and the plant lactobacillus homology of this bacterial strain reach 99%.
What cultivation Bacterium lacticum was used is MRS substratum, and culture medium prescription is: glucose 30g, peptone 10g, yeast powder 7.5g, anhydrous sodium acetate 5g, K2HPO42g, magnesium sulfate 0.8g, manganous sulfate 0.2g, dibasic ammonium citrate 2g, PH7.0-7.5, and culture temperature is 37 DEG C.
The form of bacterial classification
Plant lactobacillus cultivates 48h on substratum, generation bacterium colony figure as shown in Figure 1, colony edge is neat, smooth surface, in milk yellow; By single colonies sampling under an electron microscope (magnification is 1800 times) observe, as shown in Figure 2, cellular form is shaft-like to observations, and gramstaining is accredited as Gram-positive, without gemma and flagellum.
The present invention has found the effect of plant lactobacillus cholesterol degradation further, and is studied the effect of its cholesterol degradation.
First preparation is containing cholesterol MRS liquid nutrient medium: be first the cholesterol solution of 10gL-1 with dehydrated alcohol compound concentration, with aperture be the filtering with microporous membrane of 0.45 μm degerming after, join in the sterilizing MRS containing 0.30% Glycocholate sodium, make cholesterol ultimate density be 100mgL-1.
The preparation of fermented liquid: connect biography 3 generation in MRS, the centrifugal 10min of the 3rd culture thing 5000rmin-1, abandons supernatant liquor, washes bacterium 3 times with sterile saline is centrifugal, adjusts bacteria suspension concentration to be 3 × 109CFUmL-1.Inoculum size by 2% is inoculated in the MRS liquid nutrient medium containing cholesterol, 37 DEG C of Anaerobic culturel 24h.Retain simultaneously do not connect bacterium containing cholesterol MRS substratum, 5 DEG C of preservations, as blank.
The preparation of o-phthalaldehyde(OPA) working fluid: 50mg o-phthalaldehyde(OPA) is dissolved in 100mL Glacial acetic acid, is stirred to and dissolves completely, makes the working fluid of 0.5gL-1.This solution must use preparation on the same day.
Determination of Cholesterol Content: the centrifugal 10min of fermented liquid 5000rmin-1, retains fermentation supernatant.The physiological saline centrifuge washing 3 time of precipitation thalline with original fermented solution equivalent, washings retains.In fermented supernatant fluid and thalline washings, cholesterol level all measures by the O-phthalaldehyde method of improvement.
Concrete grammar is as follows: pipette 0.5mL sample in a clean tube, adds the ethanol of 3mL95% and the KOH of 2mL50%.Vibration mixing, saponification 10min in 60 DEG C of water-baths.After being cooled to room temperature, adding 5mL normal hexane, mix completely, extraction 1min.Add the mixing of 3mL distilled water again, left at room temperature 15min.After every separation, accurately draw 2.5mL to another clean tube from hexane layer.60 DEG C of water-baths volatilize normal hexane, add the o-phthalaldehyde(OPA) working fluid that 4mL newly prepares, and develop the color under room temperature 10min.Add the 2mL vitriol oil again, mix completely immediately, room temperature leaves standstill 10min.The light absorption value at 550nm place is measured in 90min.
Removing cholesterol rate can be drawn by following formula:
Decreasing ratio=(1-fermented supernatant fluid light absorption value/do not connect bacterium culture medium light absorption value) × 100%
The decreasing ratio calculating Bacterium lacticum cholesterol degradation is 55.19%.
Prove that a kind of plant lactobacillus of the present invention has good Degradation to cholesterol by experiment, can be used in medical aspect or healthcare products aspect.
Claims (3)
1. a plant lactobacillus, is characterized in that, described plant lactobacillus is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on December 14th, 2015, and preserving number is CGMCCNO.11875..
2. the application of plant lactobacillus according to claim 1 on cholesterol degradation.
3. plant lactobacillus according to claim 1 is for the preparation of the application on the medicine of cholesterol degradation or healthcare products.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610066292.8A CN105567601A (en) | 2016-01-29 | 2016-01-29 | Lactobacillus plantarum and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610066292.8A CN105567601A (en) | 2016-01-29 | 2016-01-29 | Lactobacillus plantarum and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105567601A true CN105567601A (en) | 2016-05-11 |
Family
ID=55878235
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610066292.8A Pending CN105567601A (en) | 2016-01-29 | 2016-01-29 | Lactobacillus plantarum and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105567601A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115873761A (en) * | 2022-11-10 | 2023-03-31 | 重庆第二师范学院 | Lactobacillus plantarum KSFY01 and application thereof |
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