CN110484460A - One plant has the lactobacillus plantarum for reducing blood fat function and its application - Google Patents
One plant has the lactobacillus plantarum for reducing blood fat function and its application Download PDFInfo
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- WPBNNNQJVZRUHP-UHFFFAOYSA-L manganese(2+);methyl n-[[2-(methoxycarbonylcarbamothioylamino)phenyl]carbamothioyl]carbamate;n-[2-(sulfidocarbothioylamino)ethyl]carbamodithioate Chemical compound [Mn+2].[S-]C(=S)NCCNC([S-])=S.COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC WPBNNNQJVZRUHP-UHFFFAOYSA-L 0.000 description 1
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/06—Antihyperlipidemics
-
- 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
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
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- 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
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/25—Lactobacillus plantarum
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- Tropical Medicine & Parasitology (AREA)
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- Nutrition Science (AREA)
- Diabetes (AREA)
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- Obesity (AREA)
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- General Chemical & Material Sciences (AREA)
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Abstract
The present invention provides one plant to have the lactobacillus plantarum for reducing blood fat function and its application, belongs to the new strains and its applied technical field of lactobacillus.Lactobacillus plantarum (Lactobacillus plantarum) nbk-MA2 of the present invention, deposit number are as follows: CGMCC No.16872.The bacterium powder and inactivated bacteria powder of lactobacillus plantarum nbk-MA2 of the present invention can reduce triglyceride in body, total cholesterol, glutamic-pyruvic transaminase, glutamic-oxalacetic transaminease content, high-density lipoprotein and low-density lipoprotein content, it makes it restore to close to normal level, has the function that reduce blood lipid；In addition, lactobacillus plantarum nbk-MA2 has good acid and Bile salt resistance, there is sufficient amount of viable bacteria that can reach enteron aisle, plays prebiotic effect.
Description
Technical field
Having the present invention relates to the new strains of lactobacillus and its applied technical field more particularly to one plant reduces blood lipid function
The lactobacillus plantarum of energy and its application.
Background technique
Dyslipidemia is one of the important risk factor of China's cardiovascular disease.Blood lipid is cholesterol, glycerol three in serum
The general name of ester and lipoid (such as phosphatide).Nearly ten years, dyslipidemia patient groups in China's are substantially improved, with national life
Horizontal raising, diet structure change, and hyperlipemia is no longer the exclusive of the elderly, and statistical data shows hyperlipemia
Morbidity crowd is developed towards rejuvenation.Especially wherein low-density lipoprotein (LDL-C) raising is to lead to artery congee to dyslipidemia
Sample hardenability cardiovascular disease (ASCVD) occurs and the key factor of development.Therefore, the generation of prevention hyperlipemia in time is to anti-
Angiocardiopathy, such as acute myocardial infarction AMI, coronary heart disease, cerebral apoplexy important in inhibiting.
In recent years, many studies have shown that lactic acid bacteria to body there are many prebiotic effect, including immunological regulation, improve allergy,
Adjust balance, enhancing immunity of organisms, the generation for preventing intestines problem and the function of reducing blood lipid of intestinal flora.
But lactic acid bacteria disclosed in the prior art reduces the insufficiency face of blood lipid, serum can be reduced by especially lacking one kind
Cholesterol, triglycerides and low-density lipoprotein, and high-density lipoprotein, glutamic-pyruvic transaminase and glutamic-oxalacetic transaminease can be effectively reduced
The lactic acid bacteria of content.
Summary of the invention
The purpose of the present invention is to provide one plant to have the lactobacillus plantarum for reducing blood fat function and its application, plant cream
Bacillus can reduce serum cholesterol, triglycerides and low-density lipoprotein and high-density lipoprotein, Gu Bingzhuan is effectively reduced
The lactic acid bacteria of adnosine deaminase and glutamic-oxalacetic transaminease content.
In order to achieve the above-mentioned object of the invention, the present invention the following technical schemes are provided:
The present invention provides one plant to have the lactobacillus plantarum (Lactobacillus plantarum) for reducing blood fat function
Nbk-MA2, deposit number are as follows: CGMCC No.16872.
The present invention also provides a kind of preparations including lactobacillus plantarum described in above scheme.
Preferably, the concentration of lactobacillus plantarum is 2 × 10 in the preparation9~6 × 1011cfu/g。
Preferably, the lactobacillus plantarum is viable bacteria or inactivated bacteria.
Preferably, the partial size of the preparation is 15~100 mesh.
The present invention also provides the preparation methods of preparation described in above scheme, comprising the following steps:
1) lactobacillus plantarum nbk-MA2 is seeded to seed culture medium, is activated, obtain primary seed solution；
2) the step 1) primary seed solution is seeded to seed culture medium, is cultivated, obtain secondary seed solution；
3) the step 2) secondary seed solution is seeded to fermentation medium, fermented, be centrifuged fermentation liquid, obtain bacterium
Body；
4) thallus described in step 3) is dried, crushed, obtain preparation.
Preferably, the seed culture medium includes the raw material of following mass percentage: 1%~2% glucose, 1.5%
~2% peptone, 0.3%~0.8% yeast extract, 0.3%~0.8% anhydrous sodium acetate, 0.2%~0.6% ammonium citrate,
0.01%~0.03% magnesium sulfate, 0.04%~0.08% manganese sulfate, 0.05%~0.15% Tween 80 and surplus water；It is described
Seed culture medium pH value is 6.5~6.8.
Preferably, the fermentation medium includes the raw material of following parts by weight: 1%~3% glucose, 1%~1.5% egg
White peptone, 0.3%~0.8% yeast extract, 0.3%~0.8% anhydrous sodium acetate, 0.1%~0.3% ammonium citrate, 0.04%
~0.08% magnesium sulfate, 0.01%~0.03% manganese sulfate, 0.05%~0.15% Tween 80 and surplus water；The fermentation training
The pH value for supporting base is 6.3~6.5.
Preferably, the temperature of the fermentation is 28~40 DEG C.
The present invention also provides lactobacillus plantarums described in above scheme or the preparation to reduce drug, the food of blood lipid in preparation
Application in product, feed or feed addictive.
Beneficial effects of the present invention: the present invention provides one plant to have the lactobacillus plantarum for reducing blood fat function
(Lactobacillusplantarum) nbk-MA2, deposit number are as follows: CGMCC No.16872.By verifying, plant of the invention
The bacterium powder and inactivated bacteria powder of object lactobacillus nbk-MA2 can significantly reduce triglyceride in body, total cholesterol, glutamic-pyruvic transaminase,
Glutamic-oxalacetic transaminease content, triglyceride, total cholesterol, high-density lipoprotein and low-density lipoprotein content, make it restore to and connect
Nearly normal level has the function that reduce blood lipid；In addition, lactobacillus plantarum nbk-MA2 has good acid and Bile salt resistance, have
Sufficient amount of viable bacteria can reach enteron aisle, play prebiotic effect.
Detailed description of the invention
Fig. 1 shows mouse serum glutamic pyruvic transminase (ALT) contents in embodiment 8 and glutamic-oxalacetic transaminease (AST) content；
Fig. 2 indicates in embodiment 9 triglyceride, total cholesterol, high-density lipoprotein and low-density lipoprotein in rat body
Content.
Biological deposits explanation
Lactobacillus plantarum (Lactobacillusplantarum) nbk-MA2 is deposited in China on December 05th, 2018
Microbiological Culture Collection administration committee common micro-organisms center, address are Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, in
Institute of microbiology, the academy of sciences, state, deposit number are as follows: CGMCC No.16872.
Specific embodiment
The present invention provides one plant to have the lactobacillus plantarum (Lactobacillus plantarum) for reducing blood fat function
Nbk-MA2, deposit number are as follows: CGMCC No.16872；The lactobacillus plantarum nbk-MA2 is divided from the Kefir granule of Tibet
From, identification obtain；The Tibet Kefir granule is in the local resident man of Tibet；The present invention does not have the isolated method
Particular determination, using this field routine strain separation method；The method of the identification is preferably molecular biology identification, more
Preferably 16S overall length is sequenced.
The present invention provides a kind of preparation including lactobacillus plantarum described in above scheme, lactobacillus plantarum in the preparation
Concentration be preferably 2 × 109~6 × 1011Cfu/g, more preferably 3 × 109~5 × 1010cfu/g；The partial size of the preparation is excellent
It is selected as 15~100 mesh, more preferably 20~80 mesh；It preferably further include protective agent and filler in the preparation.Of the invention
In specific implementation process, the lactobacillus plantarum is viable bacteria or inactivated bacteria.
The present invention also provides the preparation methods of preparation described in above scheme, comprising the following steps:
1) lactobacillus plantarum nbk-MA2 is seeded to seed culture medium, is activated, obtain primary seed solution；
2) the step 1) primary seed solution is seeded to seed culture medium, is cultivated, obtain secondary seed solution；
3) the step 2) secondary seed solution is seeded to fermentation medium, fermented, be centrifuged fermentation liquid, obtain bacterium
Body；
4) thallus described in step 3) is dried, crushed, obtain preparation.
Lactobacillus plantarum nbk-MA2 is seeded to seed culture medium first by the present invention, is activated, and first order seed is obtained
Liquid；The present invention is not particularly limited the inoculum concentration of the lactobacillus plantarum；The seed culture medium preferably includes following matter
Measure percentage raw material: 1%~2% glucose, 1.5%~2% peptone, 0.3%~0.8% yeast extract, 0.3%~
0.8% anhydrous sodium acetate, 0.2%~0.6% ammonium citrate, 0.01%~0.03% magnesium sulfate, 0.04%~0.08% sulfuric acid
The water of manganese, 0.05%~0.15% Tween 80 and surplus；It is furthermore preferred that the seed culture medium includes following mass percent
Raw material: 1.5% glucose, 1.8% peptone, 0.5% yeast extract, 0.5% anhydrous sodium acetate, 0.4% ammonium citrate,
0.02% magnesium sulfate, 0.06% manganese sulfate, 0.1% Tween 80 and surplus water；The pH value of the seed culture medium is preferably 6.5
~6.8, more preferably 6.6~6.7.In the present invention, the condition of the activation is anaerobic condition；The temperature of the activation is preferably
28~37 DEG C, more preferably 30~35 DEG C；The time of the activation is preferably 12~for 24 hours, most preferably 15~18h.
The present invention is seeded to seed culture medium after obtaining primary seed solution, by primary seed solution, is cultivated, and obtains two
Grade seed liquor；Mass percent of the inoculum concentration of the primary seed solution shared by seed culture medium is preferably 2%~8%, more
Preferably 4%~6%；The mode of the culture is preferably Anaerobic culturel；The temperature of the culture is preferably 36~38 DEG C, more excellent
It is selected as 37 DEG C；The time of the culture is preferably 13~18h, more preferably 15h；The device of the culture is preferably fermentor；
The effect of the culture is the viable count of further expansion lactobacillus plantarum, is ready for fermentation in next step.
The present invention is seeded to fermentation medium after obtaining secondary seed solution, by secondary seed solution, ferments, centrifugation hair
Zymotic fluid obtains thallus；Mass percent of the inoculum concentration of the secondary seed solution shared by fermentation medium be preferably 2%~
8%, more preferably 4%~6%；The temperature manner of the fermentation is preferably anaerobic fermentation；The temperature of the fermentation is preferably 28
~40 DEG C, more preferably 30~38 DEG C, most preferably 35~37 DEG C；The time of the fermentation is preferably 5~10h, and more preferably 6
~8h；The effect of the fermentation is the cost for significantly improving cell density, while reducing production equipment, meets and improves than production
Rate, the rival demand for reducing production cost, accelerating product ommercialization process；The revolving speed of the centrifugation is preferably 5000~
10000r/min, more preferably 6000~8000r/min；The time of the centrifugation is preferably 20~40min, more preferably 25~
35min；The temperature of the centrifugation is preferably 0~8 DEG C, and more preferably 2~6 DEG C.
In the present invention, the fermentation medium preferably includes the raw material of following mass percent: 0.5~2.5% grape
Sugar, 1.5~3% peptones, 0.5~2% yeast extract, 0.2~0.9% anhydrous sodium acetate, 0.2~0.7% ammonium citrate,
0.02~0.08% magnesium sulfate, 0.02~0.08% manganese sulfate, 0.05~0.1% Tween 80 and surplus water, it is furthermore preferred that institute
State the raw material that fermentation medium includes following mass percent: 1.5~2.5% glucose, 1.5~2.5% peptones, 1~
1.5% yeast extract, 0.3~0.6% anhydrous sodium acetate, 0.3~0.6% ammonium citrate, 0.04~0.06% magnesium sulfate, 0.04
The water of~0.06% manganese sulfate, 0.06~0.08% Tween 80 and surplus；The pH value of the fermentation medium is preferably 6.2~
7.0, more preferably 6.4~6.8, most preferably 6.5~6.6.Fermentation medium provided by the invention is suitable for heavy industrialization
Production, meets microbial nutrition demand when high density fermentation.
Viable count >=1 × 10 in specific implementation process of the present invention, in the fermentation liquid9When cfu/mL, it can stop sending out
Ferment；The measuring method of viable count is preferably according to GB4789.35-2016 " national food safety standard food in the fermentation liquid
Microbiological Test lactic acid bacteria is examined " growth curve is drawn, determined according to growth curve；The frequency of the measurement is preferably
Measurement per hour is primary after fermentation 3h.
Viable count >=1 × 10 in specific implementation process of the present invention, in the fermentation liquid9When cfu/mL, it can stop sending out
Ferment；The measuring method of viable count is preferably according to GB4789.35-2016 " national food safety standard food in the fermentation liquid
Microbiological Test lactic acid bacteria is examined " growth curve is drawn, determined according to growth curve；The frequency of the measurement is preferably
Measurement per hour is primary after fermentation 3h.
When the lactobacillus plantarum in heretofore described preparation is viable bacteria, obtained thallus is mixed with freeze drying protectant
It closes, carries out emulsification embedding, obtain emulsion；The mass ratio of the thallus and freeze drying protectant is preferably 1:0.8~1.5, more excellent
It is selected as 1:1~1.2；The freeze drying protectant preferably includes the raw material of following mass parts: 1~3 part of skimmed milk, glycerine water solution
1~5 part and 3~10 parts of water, it is furthermore preferred that the freeze drying protectant includes the raw material of following mass parts: 2 parts of skimmed milk, glycerol
2~3 parts and 5~8 parts of water of aqueous solution；The mass percentage of skimmed milk power is preferably 8%~15% in the skimmed milk, more excellent
It is selected as 10%~12%；The volumn concentration of glycerol is preferably 3% in the glycerine water solution；The skimmed milk and glycerol come
Derived from commercially available.
In the present invention, the time of the emulsification embedding is preferably 10~40min, more preferably 20~30min；The emulsification
The temperature of embedding is preferably 10~25 DEG C, and more preferably 15~20 DEG C.
In the present invention, the purpose that thallus carries out emulsification embedding is to protect the activity of lactobacillus plantarum, is reduced subsequent
Step is freeze-dried on lactobacillus plantarum viable count and active influence.
The present invention carries out precooling, vacuum freeze drying, crushing after obtaining emulsion, to emulsion, obtains preparation；Institute
The temperature for stating precooling is preferably -40~-35 DEG C, more preferably -38 DEG C；The time of the precooling is preferably 1~3h, more excellent
It is selected as 2h；The temperature of the vacuum freeze drying is preferably -40 DEG C~-10 DEG C, more preferably -30 DEG C~-20 DEG C；The vacuum
The time of freeze-drying is preferably 12~60h, more preferably 20~40h, most preferably 25~35h；The vacuum freeze drying
Vacuum degree be preferably 0.1~0.3mbar, more preferably 0.2mbar；The present invention is to the time of the crushing, temperature and equipment
It is not particularly limited, with the partial size of smashed preparation for 15~100 mesh.
The present invention also provides lactobacillus plantarums described in above scheme or the preparation to reduce drug, the food of blood lipid in preparation
Application in product, feed or feed addictive；The reduction blood lipid includes reducing serum cholesterol, triglycerides, low density lipoprotein
Albumen, high-density lipoprotein, glutamic-pyruvic transaminase and glutamic-oxalacetic transaminease content.
There is the lactobacillus plantarum for reducing blood fat function and its application to one plant provided by the invention below with reference to embodiment
It is described in detail, but they cannot be interpreted as limiting the scope of the present invention.
The screening and identification of 1 lactobacillus plantarum nbk-MA2 of embodiment
Tibet Kefir granule is acquired from the local resident man of Tibet, is diluted to appropriate ladder after sterile saline grinding is added
Degree (102~105), after using the scribing line of four zoning collimation methods on MRS agar plate, the Anaerobic culturel 48h in 37 DEG C of incubators.It chooses
Menu bacterium colony, microscopy, scribing line are repeated several times and isolate and purify to obtain lactobacillus plantarum nbk-MA2.It is to plant through molecular biology identification
Object lactobacillus is named as lactobacillus plantarum nbk-MA2.
A kind of preparation method of the bacterium powder comprising lactobacillus plantarum nbk-MA2 viable bacteria of embodiment 2
By mass percentage, seed culture medium: 1.5% glucose, 1.8% peptone, 0.5% yeast extract, 0.5%
Anhydrous sodium acetate, 0.4% ammonium citrate, 0.02% magnesium sulfate, 0.06% manganese sulfate, 0.1% Tween 80 and surplus water, pH value
6.5；
By mass percentage, fermentation medium: 2% glucose, 1.3% peptone, 0.5% yeast extract, 0.5% nothing
Water sodium acetate, 0.2% ammonium citrate, 0.06% magnesium sulfate, 0.02% manganese sulfate, 0.1% Tween 80 and surplus water, pH value is
6.4；
In parts by mass, freeze drying protectant: 10% 2 parts of skimmed milk, 2 parts of 3% glycerol and 5 parts of water.
Preparation method
1) lactobacillus plantarum is inoculated into activation (37 DEG C, 20h) in the test tube equipped with seed culture medium, obtains first order seed
Liquid；
2) primary seed solution is forwarded in seed culture medium according to the inoculum concentration that mass percent is 4% and obtains second level kind
Sub- liquid, condition of culture are 37 DEG C, Anaerobic culturel 15h；
3) secondary seed solution is accessed in fermentation medium and carries out high density fermentation, secondary seed solution is connected to fermentation medium
In inoculum concentration be 4% (mass percent), condition of culture be 37 DEG C, Anaerobic culturel 8h, reach 1 to cell density in fermentation liquid
×109Cfu/mL, collects fermentation liquid, and centrifugation (8000r/min, 30min) obtains thallus；
4) protective agent (thallus: protective agent mass ratio is 1:1) is added in thallus and carries out emulsification embedding, emulsification times 25min；
5) by emulsion at -38 DEG C precooling 2h, by after precooling emulsion carry out (- 40 DEG C of vacuum freeze drying
To 30 degrees Celsius, time 30h), vacuum degree 0.2mbar；It is crushed to 20 mesh after vacuum freeze drying, obtains preparation, institute
The viable count for obtaining preparation is 5.8 × 1011cfu/g。
A kind of preparation method of the inactivated bacteria powder comprising lactobacillus plantarum nbk-MA2 inactivated bacteria of embodiment 3 presses quality percentage
Than meter, seed culture medium: 1.5% glucose, 1.8% peptone, 0.5% yeast extract, 0.5% anhydrous sodium acetate, 0.4% lemon
Lemon acid ammonium, 0.02% magnesium sulfate, 0.06% manganese sulfate, 0.1% Tween 80 and surplus water, pH value 6.8；
By mass percentage, fermentation medium: 2% glucose, 1.3% peptone, 0.5% yeast extract, 0.5% nothing
Water sodium acetate, 0.2% ammonium citrate, 0.06% magnesium sulfate, 0.02% manganese sulfate, 0.1% Tween 80 and surplus water, pH value is
6.4。
Preparation method:
1) lactobacillus plantarum is inoculated into activation (37 DEG C, 20h) in the test tube equipped with seed culture medium, obtains first order seed
Liquid；
2) primary seed solution is forwarded in seed culture medium according to the inoculum concentration that mass percent is 4% and obtains second level kind
Sub- liquid, condition of culture are 37 DEG C, Anaerobic culturel 15h；
3) secondary seed solution is accessed in fermentation medium and carries out high density fermentation, secondary seed solution is connected to fermentation medium
In inoculum concentration be 4% (mass percent), condition of culture be 37 DEG C, Anaerobic culturel 8h reaches to cell density in fermentation liquid
109Cfu/mL, collects fermentation liquid, and centrifugation (8000r/min, 30min) obtains thallus；
4) thallus is dried, drying temperature is 45 DEG C (2.5d)；It is crushed to 20 mesh after drying, obtains plant cream
Bacillus nbk-MA2 inactivated bacteria powder.
The outer cholesterol-lowering activity research of the lactobacillus plantarum nbk-MA2 bacterium powder of 4 embodiment 2 of embodiment
The obtained lactobacillus plantarum nbk-MA2 bacterium powder of embodiment 2 is inoculated in cholesterol by the inoculum concentration of 2% (w/v)
Content is in the MRS-CHO culture medium of 30mg/mL, 37 DEG C of Anaerobic culturels for 24 hours after, it is solid to measure gallbladder in supernatant for centrifuging and taking supernatant
The content of alcohol is 13.5mg/mL, and the removal rate of cholesterol, calculated result are as follows: the removal rate of cholesterol are calculated according to following formula
It is 55%.
The outer cholesterol-lowering activity research of the lactobacillus plantarum nbk-MA2 inactivated bacteria powder of 5 embodiment 3 of embodiment
It is 30mg/ that the obtained lactobacillus plantarum nbk-MA2 inactivated bacteria powder of 0.1g embodiment 3, which is put into cholesterol level,
It in the aqueous solution of mL, is placed in 37 DEG C of environment and places 8h, centrifuging and taking supernatant, measuring the content of cholesterol in supernatant is
9.9mg/mL calculates the removal rate of cholesterol, calculated result according to the formula in embodiment 4 are as follows: the removal rate of cholesterol is
67%.
The lactobacillus plantarum nbk-MA2 bacterium powder acid-resistant property of 6 embodiment 2 of embodiment is studied
The nbk-MA2 bacterium powder that embodiment 2 obtains is connected to respectively with the inoculum concentration of 1% (w/v) to pH is 6.0 and pH is 2.0
MRS culture medium in cultivate 3h after using colony counting method carry out count plate, as a result referring to table 1.
The lactobacillus plantarum nbk-MA2 bacterium powder acid-resistant property result of study of 1 embodiment 2 of table
As can be seen that lactobacillus plantarum nbk-MA2 still has sufficient amount of viable bacteria can after 3h under conditions of 2.0 pH
To pass through stomach environment.
The bile tolerance characteristic of the lactobacillus plantarum nbk-MA2 bacterium powder of 7 embodiment 2 of embodiment
The nbk-MA2 bacterium powder that embodiment 2 obtains is connected to gallbladder salinity with the inoculum concentration of 1% (w/v) respectively as 0% (w/
V) and gallbladder salinity is carries out count plate using colony counting method after culture 3h in 0.4% MRS culture medium, as a result referring to table
2。
The bile tolerance characteristic result of study of the lactobacillus plantarum nbk-MA2 bacterium powder of 2 embodiment 2 of table
As can be seen that lactobacillus plantarum nbk-MA2 has preferable Bile salt resistance under 0.4% gallbladder salinity.
The lactobacillus plantarum nbk-MA2 bacterium powder of 8 embodiment 2 of embodiment and the lactobacillus plantarum nbk-MA2 inactivation of embodiment 3
Bacterium powder is reducing the application in lipid of mice content
1. test material:
Experimental animal: SPF grades health Balb/c male mice 50,18~22g of weight.
Subjects:
Basal feed: being purchased from Si Beifu (Beijing) Biotechnology Co., Ltd, and trade name rats and mice maintains feed；
MCD feed: it is purchased from Beijing Biotechnology Co., Ltd, Mao Sibei section.
With the lactobacillus plantarum nbk-MA2 bacterium powder of embodiment 2 and the lactobacillus plantarum nbk-MA2 inactivated bacteria powder of embodiment 3
For test specimen
1.2 modelings and grouping:
Modeling: in addition to blank group, remaining each group mouse was using MCD forage feed 2 weeks, as follows by 5 groups of mouse
Carry out stomach-filling:
A blank group: blank control group ad lib basal feed, daily stomach-filling 0.1mL/10g.bw physiological saline, continuously
30d；
B model group: model group mouse ad lib MCD feed, daily stomach-filling 0.1mL/10g.bw physiological saline, continuously
30d；
C lactobacillus plantarum nbk-MA2 bacterium powder group: weighing 0.01g lactobacillus plantarum nbk-MA2 bacterium powder described in embodiment 1,
It is dissolved in 580mL sterile saline to get stomach-filling solution, takes the daily stomach-filling mouse of the stomach-filling solution, given low is
0.1mL/10g.bw, continuous 30d；
D lactobacillus plantarum nbk-MA2 inactivated bacteria powder group: 0.01g lactobacillus plantarum nbk-MA2 as described in example 2 is weighed
Bacterium powder is dissolved in 580mL sterile saline to get stomach-filling solution, takes the daily stomach-filling mouse of the stomach-filling solution, and given low is
0.1mL/10g.bw, continuous 30d.
After experiment, after being deprived of food but not water 12h, hematometry mice serum glutamic-pyruvic transaminase (ALT) content and millet straw are taken
Transaminase (AST) content.Experimental result is as shown in Figure 1 (* is indicated compared to model group P < 0.05).
By Fig. 1 result it is found that the lactobacillus plantarum nbk-MA2 bacterium powder of embodiment 2 and the lactobacillus plantarum nbk- of embodiment 3
MA2 inactivated bacteria powder can significantly reduce the content of glutamic-pyruvic transaminase and glutamic-oxalacetic transaminease in model mice serum.Glutamic-pyruvic transaminase and
Glutamic-oxalacetic transaminease is the important indicator of liver function, lactobacillus plantarum can significantly reduce in model mice serum glutamic-pyruvic transaminase and
The content of glutamic-oxalacetic transaminease can prevent the fatty liver caused by hyperlipidemia, mitigate damage of the hyperlipidemia to liver
The lactobacillus plantarum nbk-MA2 bacterium powder of 9 embodiment 2 of embodiment and the lactobacillus plantarum nbk-MA2 inactivation of embodiment 3
Bacterium powder is reducing the application in rat fat content
1. test material:
Experimental animal: SPF grades healthy SD male rat 40,180~220g of weight.
Subjects:
Basal feed: being purchased from Si Beifu (Beijing) Biotechnology Co., Ltd, and trade name rats and mice maintains feed；
High lipid food (%): normal diet 80, lard 16, cholesterol 2, Pig cholate 2.
With the lactobacillus plantarum nbk-MA2 bacterium powder of embodiment 2 and the lactobacillus plantarum nbk-MA2 inactivated bacteria powder of embodiment 3
For test specimen
1.2 modelings and grouping:
Modeling: in addition to blank group, remaining each group rat was using high lipid food nursing 4 weeks, as follows by 5 groups of mouse
Carry out stomach-filling:
A blank group: blank control group ad lib basal feed, daily stomach-filling 0.1mL/10g.bw physiological saline, continuously
30d；
B model group: model group rats ad lib high lipid food, daily stomach-filling 0.1mL/10g.bw physiological saline, continuously
30d；
C lactobacillus plantarum nbk-MA2 bacterium powder group: weighing 0.01g lactobacillus plantarum nbk-MA2 bacterium powder described in embodiment 1,
It is dissolved in 580mL sterile saline to get stomach-filling solution, takes the daily stomach-filling rat of the stomach-filling solution, given low is
0.1mL/10g.bw, continuous 30d；
D lactobacillus plantarum nbk-MA2 inactivated bacteria powder group: 0.01g lactobacillus plantarum nbk-MA2 as described in example 2 is weighed
Bacterium powder is dissolved in 580mL sterile saline to get stomach-filling solution, takes the daily stomach-filling rat of the stomach-filling solution, and given low is
0.1mL/10g.bw, continuous 30d.
After experiment, after being deprived of food but not water 12h, take triglyceride (TG) in hematometry rat body, total cholesterol (TG),
The content of high-density lipoprotein (HDL-C), low-density lipoprotein (LDL-C).Experimental result is referring to fig. 2.Fig. 2 indicates embodiment 2
Lactobacillus plantarum nbk-MA2 bacterium powder and embodiment 3 lactobacillus plantarum nbk-MA2 inactivated bacteria powder rat body in TG, TC, HDL-
The influence of C and LDL-C content；Note: letter is identical between homologous series, shows that difference is not significant, and letter difference then shows that difference is aobvious
It writes, p < 0.05.
By Fig. 2 result it is found that the lactobacillus plantarum nbk-MA2 bacterium powder of embodiment 2 and the lactobacillus plantarum nbk- of embodiment 3
MA2 inactivated bacteria powder can significantly reduce triglyceride, total cholesterol, high-density lipoprotein and low-density lipoprotein in rat body and contain
Amount, makes it restore to close to normal level, has the function that reduce blood lipid.
Application of the lactobacillus plantarum nbk-MA2 bacterium powder of 10 embodiment 2 of embodiment in preparation fermentation mixed fruit and vegetable juice
By mass fraction percentages, 10% concentrated apple juice, 10% concentrated pineapple juice, 0.75% oligofructose, 0.5%
Stachyose, 45.5% softened water.Inoculating starter, lactobacillus plantarum nbk-MA2: lactobacillus bulgaricus and streptococcus thermophilus
Viable count ratio is 10:1:1, and inoculum concentration is 1 × 107CFU/mL.Mix, 37 DEG C of anaerobic fermentations for 24 hours, when pH value reaches 3.37,
Terminate fermentation；100 DEG C of sterilizing 15min, sterile filling is to get fermented fruits and vegetables juice drink.
The lactobacillus plantarum nbk-MA2 inactivated bacteria powder of 11 embodiment 3 of embodiment is preparing solid with blood fat reducing function
Application in beverage
Lactobacillus plantarum inactivated bacteria powder 5%, oligofructose 10%, xylo-oligosaccharide 10%, erythrose prepared by embodiment 3
Alcohol 32%, resistant dextrin 40% and soybean peptide 3% are mixed, and the composite probiotics preparations with hypolipidemic activity are made.
As seen from the above embodiment, the present invention provides one plant to have the lactobacillus plantarum nbk-MA2 for reducing blood fat function,
The bacterium powder of lactobacillus plantarum nbk-MA2 can significantly reduce triglyceride in body, total cholesterol, glutamic-pyruvic transaminase and millet straw and turn
Adnosine deaminase content makes it restore to close to normal level, has the function that reduce blood lipid；Secondly, lactobacillus plantarum nbk-MA2 bacterium powder
And its inactivated bacteria powder can significantly reduce triglyceride in body, total cholesterol, high-density lipoprotein and low-density lipoprotein content,
It makes it restore to close to normal level, has the function that reduce blood lipid；In addition, lactobacillus plantarum nbk-MA2 have it is good acid and
Bile salt resistance has sufficient amount of viable bacteria that can reach enteron aisle, plays prebiotic effect.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (10)
1. one plant has lactobacillus plantarum (Lactobacillus plantarum) nbk-MA2 for reducing blood fat function, preservation is compiled
Number are as follows: CGMCC No.16872.
2. a kind of preparation including lactobacillus plantarum described in claim 1.
3. preparation according to claim 2, which is characterized in that the concentration of lactobacillus plantarum is 2 × 10 in the preparation9~6
×1011cfu/g。
4. preparation according to claim 2 or 3, which is characterized in that the lactobacillus plantarum is viable bacteria or inactivated bacteria.
5. preparation according to claim 4, which is characterized in that the partial size of the preparation is 15~100 mesh.
6. the preparation method of preparation described in claim 2~5 any one, comprising the following steps:
1) lactobacillus plantarum nbk-MA2 is seeded to seed culture medium, is activated, obtain primary seed solution；
2) the step 1) primary seed solution is seeded to seed culture medium, is cultivated, obtain secondary seed solution；
3) the step 2) secondary seed solution is seeded to fermentation medium, fermented, be centrifuged fermentation liquid, obtain thallus；
4) thallus described in step 3) is dried, crushed, obtain preparation.
7. preparation method according to claim 6, which is characterized in that the seed culture medium includes that following quality percentage contains
The raw material of amount: 1%~2% glucose, 1.5%~2% peptone, 0.3%~0.8% yeast extract, 0.3%~0.8% nothing
Water sodium acetate, 0.2%~0.6% ammonium citrate, 0.01%~0.03% magnesium sulfate, 0.04%~0.08% manganese sulfate,
The water of 0.05%~0.15% Tween 80 and surplus；
The seed culture medium pH value is 6.5~6.8.
8. preparation method according to claim 6, which is characterized in that the fermentation medium includes the original of following parts by weight
Material: 1%~3% glucose, 1%~1.5% peptone, 0.3%~0.8% yeast extract, 0.3%~0.8% anhydrous acetic acid
Sodium, 0.1%~0.3% ammonium citrate, 0.04%~0.08% magnesium sulfate, 0.01%~0.03% manganese sulfate, 0.05%~
The water of 0.15% Tween 80 and surplus；
The pH value of the fermentation medium is 6.3~6.5.
9. preparation method according to claim 6, which is characterized in that the temperature of the fermentation is 28~40 DEG C.
10. preparation described in lactobacillus plantarum described in claim 1 or claim 2~5 any one reduces blood lipid in preparation
Application in drug, food, feed or feed addictive.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111000246A (en) * | 2019-12-27 | 2020-04-14 | 汤臣倍健股份有限公司 | Probiotic dietary fiber composition for assisting in reducing triglyceride, application thereof and health-care product |
| CN111793577A (en) * | 2020-07-02 | 2020-10-20 | 重庆第二师范学院 | Lactobacillus plantarum with weight-losing and lipid-lowering functions and application thereof |
| CN112813135A (en) * | 2021-04-07 | 2021-05-18 | 河北一然生物科技有限公司 | Method for detecting cheese strains mixed in lactobacillus plantarum |
| CN112933116A (en) * | 2021-04-14 | 2021-06-11 | 江南大学 | Application of lactobacillus plantarum in preparation of vascular protective agent |
| CN114869916A (en) * | 2022-03-31 | 2022-08-09 | 善恩康生物科技（苏州）有限公司 | Lactobacillus plantarum metazoan and application thereof in intestinal microecology modification and cholesterol reduction products |
| CN115105535A (en) * | 2022-03-08 | 2022-09-27 | 润盈生物工程（上海）有限公司 | Application of lactobacillus plantarum Lp-G18 in regulating immunity and reducing blood fat of children |
-
2019
- 2019-02-19 CN CN201910121419.5A patent/CN110484460A/en not_active Withdrawn
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111000246A (en) * | 2019-12-27 | 2020-04-14 | 汤臣倍健股份有限公司 | Probiotic dietary fiber composition for assisting in reducing triglyceride, application thereof and health-care product |
| CN111793577A (en) * | 2020-07-02 | 2020-10-20 | 重庆第二师范学院 | Lactobacillus plantarum with weight-losing and lipid-lowering functions and application thereof |
| CN111793577B (en) * | 2020-07-02 | 2022-02-11 | 重庆第二师范学院 | Lactobacillus plantarum with weight-losing and lipid-lowering functions and application thereof |
| CN112813135A (en) * | 2021-04-07 | 2021-05-18 | 河北一然生物科技有限公司 | Method for detecting cheese strains mixed in lactobacillus plantarum |
| CN112933116A (en) * | 2021-04-14 | 2021-06-11 | 江南大学 | Application of lactobacillus plantarum in preparation of vascular protective agent |
| CN112933116B (en) * | 2021-04-14 | 2024-01-12 | 江南大学 | Application of lactobacillus plantarum in preparation of vascular protecting agent |
| CN115105535A (en) * | 2022-03-08 | 2022-09-27 | 润盈生物工程（上海）有限公司 | Application of lactobacillus plantarum Lp-G18 in regulating immunity and reducing blood fat of children |
| CN114869916A (en) * | 2022-03-31 | 2022-08-09 | 善恩康生物科技（苏州）有限公司 | Lactobacillus plantarum metazoan and application thereof in intestinal microecology modification and cholesterol reduction products |
| WO2023185941A1 (en) * | 2022-03-31 | 2023-10-05 | 善恩康生物科技（苏州）有限公司 | Lactobacillus plantarum postbiotic and use thereof in products for changing intestinal microecology and reducing cholesterol |
| GB2620891A (en) * | 2022-03-31 | 2024-01-24 | Thankcome Biological Science And Tech Suzhou Co Ltd | Lactobacillus plantarum postbiotic and use thereof in products for changing intestinal microecology and reducing cholesterol |
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