CN109456909A - One plant has the Lactobacillus helveticus for reducing cholesterol ability - Google Patents
One plant has the Lactobacillus helveticus for reducing cholesterol ability Download PDFInfo
- Publication number
- CN109456909A CN109456909A CN201810941393.4A CN201810941393A CN109456909A CN 109456909 A CN109456909 A CN 109456909A CN 201810941393 A CN201810941393 A CN 201810941393A CN 109456909 A CN109456909 A CN 109456909A
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- CN
- China
- Prior art keywords
- lactobacillus helveticus
- ability
- bacterial strain
- lactic acid
- cholesterol
- 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.)
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Classifications
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- 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
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- A—HUMAN NECESSITIES
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- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
- A23C9/1234—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt characterised by using a Lactobacillus sp. other than Lactobacillus Bulgaricus, including Bificlobacterium sp.
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
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- A—HUMAN NECESSITIES
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/06—Antihyperlipidemics
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/02—Separating microorganisms from their culture media
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract
The invention discloses one plant to have the Lactobacillus helveticus for reducing cholesterol ability, deposit number CGMCC15778.It is described that there is the Lactobacillus helveticus for reducing cholesterol ability to isolate from Qinghai-Tibet traditional zymotic yak yoghourt, serum total cholesterol, triglycerides, low-density lipoprotein can be significantly reduced, with stronger bacteriostasis, there is certain rejection ability to four kinds of Escherichia coli, staphylococcus aureus, listeria spp and salmonella typhimurium pathogenic bacteria, can be applied to fermented food preparation and the preparation of health care product and drug.
Description
Technical field
The present invention relates to field of biotechnology more particularly to functional lactobacillus exploitation and application field, especially one plant
With the Lactobacillus helveticus for reducing cholesterol ability.
Background technique
Blood cholesterol levels are excessively a principal elements for causing cardiovascular disease.It is reported that the cholesterol in serum drops
Low 1% can make cardiovascular disease reduce by 2% to 3% (Manson et al, 1992).Pass through drug therapy treatment hyperlipidemia
Not only price is more expensive but also can generate side effect to human health for disease.It can be dropped for this reason, inquiring into and supplementing one kind in the diet
The substance of low serum cholesterol causes the great interest of scientist.In recent years, lactic acid bacteria can potentially reduce gallbladder as one kind
The dairy produce additive of sterol obtained extensive concern (Chandan et al., 1999；Nicole et al.,2000).Greatly
Quantifier elimination shows, lactic acid bacteria can be effectively reduced the cholesterol in serum, the content of triglycerides and low-density lipoprotein,
And lactic acid bacteria can reduce cholesterol this function confirmed on people, mouse and pig (Kawase etc., 2000；
Haberer etc., 2003；Sadrzadeh etc., 2010).From the current study, lactic acid bacteria can reduce blood cholesterol levels
Partly cause is that lactic acid bacteria can generate a kind of cholate hydrolase so that conjucated bile acids salt (Ha is removed in the formation of hydrolyzable cholate
Deng 2006).Because this go conjucated bile acids salt that can be easier that (Kumar etc., 2011) is discharged with excrement than conjucated bile acids salt,
To reduce the content of cholesterol in serum by the excretion for increasing cholate.Can thus it make from cholesterol biosynthesis
The amount of cholate increases, or reduce cholesterol dissolubility finally reduce amount that cholesterol absorbed by alimentary canal (Huang etc.,
2013)。
The lactic acid bacteria strains that research institute in relation to lactic acid bacteria norcholesterol utilizes are all from low altitude area environment fermented dairy product
In isolated lactic acid bacteria.And the norcholesterol ability of current lactic acid bacteria is lower, such as Yin Jun rosy clouds etc. (2008) are from fermentation
One plant of preferable Pediococcus acidilactici of norcholesterol ability, degradation in vitro 47.58% are isolated in vegetables.Lu Xiaona etc.
(2006) isolated 10 lactobacillus for degrading cholesterol, degradation rate are up to 61.7% from the koumiss of Xinjiang.Occupy magnificent (2007)
An isolated lactobacillus plantarum from Yoghourt, which is 48.1% to the cholesterol clearance rate in chemical combination source, to yolk
The clearance rate of source cholesterol is 58.2%.In addition, there are also the reports that lactobacillus for degrading cholesterol is separated from other fermented foods, but
Degradation rate is not high.Therefore, there is the lactic acid bacteria of high norcholesterol ability to have great importance for screening.
The fermentation of lactic acid bacteria has its special living environment in Qinghai-Tibet traditional zymotic yak yoghourt.Research shows that
There is the characteristic of high lipid content for the yak milk of traditional zymotic yak yoghourt, the content of fat is about fatty in milk cow milk
1 times or more (Guo etc. 2014).It is can be found that by observing the Yoghourt that local herdsman makes, the Yoghourt top in fermented yoghourt container
Layer is usually constructed with a thick layer fat deposit.For this reason, we guess fermentation yak yoghourt cream of the long term survival in environment high in fat
Sour bacterium may produce stronger tolerance to environment high in fat, and have compared with high cholesterol hydrolysing activity.Therefore traditional zymotic
Yak yoghourt is for a kind of special material for screening high cholesterol-lowering activity lactic acid bacteria.
Summary of the invention
The purpose of the present invention is to provide a kind of plateau Lactobacillus helveticus, have high norcholesterol, triglycerides, low-density
The effects of lipoprotein and Liver fatty deposition.
To achieve the goals above, present invention screening lactobacillus from Qinghai-Tibet traditional zymotic yak yoghourt, step is such as
Under:
(1) isolate and purify: the microorganism in Qinghai-Tibet traditional zymotic yak yoghourt passes through MRS, M17, KFS and SL tetra-
The different culture medium culture of kind, selects feature bacterial strain and purifies, obtain pure isolated bacterial strain.
(2) bacterial strain is identified: in conjunction with hydrogen peroxide feminine gender, Gram-positive, thalli morphology and microscopy separating, selecting
Like lactic acid bacteria strains, 16S rDNA sequencing is carried out, Blast comparison is carried out in NCBI, identifies and save 720 strains of lactic acid bacteria bacterial strains.
(3) preliminary screening: by the drop of the lactic acid bacteria strains identified in o-phthalaldehyde colorimetric method for determining step (2)
Cholesterol ability filters out the ability that bacterial strain BX95 of the present invention has high norcholesterol, is accredited as Lactobacillus helveticus in (2)
(Lactobacillus helveticus)。
(4) the 16S rRNA gene order of bacterial strain BX95 is registered in Genbank, obtains Genbank database bacterium
The sequence number of strain BX95: MF093231.
The Lactobacillus helveticus with high norcholesterol is applied in preparing fermented dairy product.
The Lactobacillus helveticus with high norcholesterol is applied in preparing health food and drug.
Lactobacillus helveticus of the present invention with norcholesterol is deposited in China Microbiological bacterium on May 21st, 2018
Kind preservation administration committee common micro-organisms center, address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, the Chinese Academy of Sciences
Institute of microbiology, deposit number CGMCC15778.
It is very good that the present invention provides the above-mentioned Rui Shi lactobacillus acid-fast ability with norcholesterol ability, hydrophobic ability and right
Performance is good in terms of the tolerance of artificial gastro-intestinal Fluid.It can apply in preparing fermented dairy product and health care product and drug.
The above-mentioned Lactobacillus helveticus with norcholesterol ability has very strong norcholesterol ability, much higher than existing
Bacterial strain, and it can be substantially reduced deposition fatty in hyperlipidemia model SD rat liver, hyperlipidemia model SD rat blood serum total cholesterol,
Triglycerides and low-density lipoprotein, and to Escherichia coli, staphylococcus aureus, listeria spp and salmonella typhimurium
Four kinds of pathogenic bacteria have certain rejection ability, can be used in food fermentation and health care product and drug, can reduce serum gallbladder
Sterol and Liver fatty deposition inhibit pathogen infection, improve immunity of organisms.
Detailed description of the invention
Fig. 1 is survival rate of the BX95 of the present invention in different incubation time points；
Fig. 2 is bacterium number of the BX95 of the present invention in different incubation time points；
Fig. 3 is change of moisture content comparison diagram in SD rat excrement in embodiment 5；
Fig. 4 is the 8th week serum total cholesterol content balance figure of SD rat in embodiment 5；
Fig. 5 is the 8th week serum triglyceride content balance figure of SD rat in embodiment 5；
Fig. 6 is the 8th week serum low-density LP comparison diagram of SD rat in embodiment 5；
Fig. 7 is the 8th week serum high-density LP comparison diagram of SD rat in embodiment 5；
Fig. 8 is blank control group SD rat liver tissue slice map in embodiment 5；
Fig. 9 is high in fat group of SD rat liver tissue slice map in embodiment 5；
Figure 10 is high in fat+Lactobacillus helveticus BX95 group SD rat liver tissue slice map in embodiment 5.
Specific embodiment
Below in conjunction with specific embodiment, and referring to attached drawing, the present invention is described in further details.
Cause of disease bacteria strain used in 1 bacterial strain screening culture medium of embodiment and bacteriostatic test
One, related culture medium prescription
MRS culture medium is the basal medium for cultivating lactic acid bacteria, and M17 culture medium and SL culture medium are separating lactic acid bacterium
Culture medium, wherein SL culture medium is the culture medium for separating lactobacillus acidoilus.
Lactic acid bacteria culture medium MRS: peptone 10g, beef extract 10g, yeast extract 5g, diammonium hydrogen citrate 2g, Portugal
Grape sugar 20g, Tween-80 1mL, sodium acetate 5g, dipotassium hydrogen phosphate 2g, magnesium sulfate 0.58g, manganese sulfate 0.25g, agar 15g steam
Distilled water 1000mL, pH 6.2-6.6.Preparation method: will be added to the water heating for dissolving by all the components in addition to agar, and adjust pH 6.2~6.4,
Agar, 121 DEG C of sterilizing 15min, while hot inverted plate is added.
M17 culture medium: it accurately weighs and plants matter peptone 5.0g, yeast extract 5.0g, polyprotein peptone 5.0g, Vitamin C
Sour 0.5g, beef extract 2.5g, β-phosphoglycerol disodium 19g measure 1.0mol/L MgSO47H2O 1.0mL, distilled water
1000mL, agar 15g.Preparation method: will be added to the water heating for dissolving by all the components in addition to agar, and adjust pH 7.0, be added agar, and 121
DEG C sterilizing 15min, while hot inverted plate.
SL culture medium: weighing casein hydrolysate 10g, yeast extract 5g, diammonium hydrogen citrate 2 g, sodium acetate 25g,
Epsom salt 0.58g, agar 15g, glucose 20g, Tween 80 1.0mL, dipotassium hydrogen phosphate 6g, ferrous sulfate heptahydrate
0.03g, four water manganese sulfate 0.15g, distilled water 1000mL.Preparation method: will be added to the water heating for dissolving by all the components in addition to agar, adjust
PH 5.4, the agar dissolved by heating are uniformly mixed, boil 5min while stirring, be cooled to 50 DEG C, inverted plate.
KFS streptococcus selective medium: proteose peptone 10g, yeast powder 10g, sodium glycero-phosphate 10g, sodium chloride
5g, maltose 20g, lactose 1g, Sodium azide 0.4g, agar 13g, bromocresol purple 0.015g adjust pH 7.2.Preparation method: fine jade will be removed
The outer all the components of rouge are added to the water heating for dissolving, adjust pH 7.2, agar is added, 121 DEG C of sterilizing 15min are cooled to 50-60 DEG C
When, sterile 1%TTC solution 10mL is added, mixes, while hot inverted plate.
Nutrient medium (NB): peptone 10g, beef extract 3g, sodium chloride 5g, distilled water 1000 mL, pH 7.0.Preparation method:
Mentioned component is mixed, pH, 121 DEG C of sterilizing 15min are adjusted after dissolution, cooling is spare.When preparing nutrient medium plate, it is added
0.8% agar.
Brain heart infusion broth (BHI): peptone 10g is dehydrated small bovine brain leaching powder 12.5g, is dehydrated beef heart infusion 5g, sodium chloride
5g, glucose 2g, disodium hydrogen phosphate 2.5g, distilled water 1000mL, pH 7.4.Preparation method: mentioned component is mixed, and is adjusted after dissolution
PH, 121 DEG C of sterilizing 15min, cooling are spare.When preparing culture medium flat plate, 0.8% agar is added.
Two, cause of disease bacteria strain used in bacteriostatic test
Standard pathogenic bacteria: salmonella typhimurium (Salmonella typhimurium, CICC10420), it is single to increase Li Si
Special Salmonella (Listeria monocytogenes, CICC21583), staphylococcus aureus (Staphylococcus
Aureus, CICC10384), escherichia coli (Escherichia coli, CICC20234).Single listeria spp that increases is used
Brain-heart infusion medium culture, the nutrient medium culture of remaining pathogenic bacteria.
Embodiment 2: it is separated from Qinghai-Tibet traditional zymotic yak yoghourt and identifies lactic acid bacteria
One, sample source
From the 720 of Qinghai-Tibet 6 areas (Nagqu, Maqu, Hongyuan, cajaput, the day Zhuhe Ruoergai) acquisition in China
Part traditional zymotic yak yoghourt.
Two, the isolated lactic acid bacteria from traditional yak dairy products
It takes 1mL fermentation yak dairy products in 1:10 ratio, is diluted with sterile saline, this is 10-1Dilution.Then it inhales
The above-mentioned dilution of 1mL, then 10 times of dilutions are taken, are 10-2Dilution, and so on.Choose 10-5、10-6、10-7Three dilutions, point
100 μ L dilutions are not drawn to be equably coated on respectively on MRS, M17, KFS and SL (adjusting pH5.4) plating medium, are existed respectively
37 DEG C, 42 DEG C, 42 DEG C and 37 DEG C anaerobism constant temperature incubation 48 hours, picking feature bacterium colony, and being further purified obtains pure separation
Lactic acid bacteria strains.
Three, bacterial strain is identified
(1) Gram's staining, Catalase determination
Gram's staining: the pure bacterium colony on picking plate carries out smear, fixation, crystal violet, and just dye, iodine solution mordant dyeing, alcohol are de-
Color, sarranine redye, dry, oil mirror microscopy.Bluish violet is gram-positive bacteria, and red is Gram-negative bacteria.
Catalase determination: exposing 30min for bacterium to be measured in air, draws a small amount of 3% (volume fraction) with dropper
Hydrogen peroxide drips the bacterium colony grown in planar surface, and it is the positive that observation, which has bubble generation, after 2-3 minutes, and inaction is negative.
The bacterial strain Gram's staining is the positive, and hydrogen peroxide determination is feminine gender, can tentatively be considered as lactic acid bacteria.
(2) 16SrRNA is identified
After lactic acid bacteria after purification cultivates 8h in MRS culture medium, 10000rpm/min is centrifuged 3-5 min and collects thallus,
DNA is extracted according to kit application method.Then PCR amplification is carried out, in universal primer 27F and 1492R (the Monis et of bacterium
Al, 2005) under amplification, the amplification of 16SrRNA is carried out.Amplification condition: 94 DEG C of 3min；94 DEG C of 30s, 55 DEG C of 30s, 72 DEG C
1min；72 DEG C of 5min, 30 circulations.PCR product serves Hai Meijisheng sequencing company and carries out sequence analysis.Lactic acid bacteria will be measured
Bacterial strain 16S rRNA gene order is searched in GenBank with BLAST (http://wwwncbi.nlm.nih.gov/blast/)
Rope is greater than 98% with strain to be tested homology score, then it is believed that they belong to same kind.Totally 72 plants of appraisable lactic acid bacteria,
It is completely used for the screening of high lactobacillus for degrading cholesterol.
Embodiment 3: the screening of high lactobacillus for degrading cholesterol
One, the preparation of water-soluble cholesterol solution and lactic acid bacteria
Prepare water-soluble cholesterol solution, in superclean bench membrane filtration degerming.After lactic acid bacteria sample activates 3-4 generation
(the last time activation culture time is 12-16h), adjusts the quantity of bacterium to 1.0 × 109 cfu/mL, by the inoculation of 1% (v/v)
Amount is inoculated in the MRS-Thio culture medium of the bilein containing 0.3%, and the additive amount of water-soluble cholesterol is 100 μ L/mL.Blank
Control is the MRS-Thio culture medium for not adding the containing water-soluble cholesterol of bacterium solution.1mL, 10000rpm/min, 2min are sampled, is taken
Supernatant 0.5mL, using the cholesterol level in culture solution after o-phthalaldehyde method measurement inoculation.Remaining fermentation liquid is in 37 DEG C
Stationary culture is for 24 hours.
Two, the measurement of cholesterol concentration
Using the content (Rudel L L et al, 1973) of o-phthalaldehyde method measurement cholesterol.To 7 with plug
Cholesterol standard liquid and dehydrated alcohol as shown in table 2-3 are accurately drawn in test tube, are dried with nitrogen, are rapidly added 4mL neighbour's benzene two
Formalin, vortex 1min, is placed at room temperature for 10min, is slowly added to the concentrated sulfuric acid of 2 mL, and vortex 1min is uniformly mixed, and room temperature is put
10min is set, cholesterol OD550 nm light absorption value is measured.Using the light absorption value of OD550nm as ordinate, the concentration of standard solution of cholesterol
For abscissa, standard curve is drawn.
1 cholesterol standard curve plotting of table
Streptococcus acidi lactici fermented solution derived above is carried out to the measurement of cholesterol level, step according to the mensuration program of standard curve
Suddenly it is changed (Gilliland, S.E.et al, 1985).Bacterium solution 1mL, 10000rpm/min, 2min after taking culture 48h.
Accurate Aspirate supernatant 0.5mL adds the KOH of 2mL 33% to be sufficiently mixed uniformly, the second of 3mL 95% in the test tube with plug
Alcoholic solution, vortex 1min, jumps a queue, 60 DEG C of water-bath 10min, cooling.Add 5mL n-hexane vortex 1min.Distilled water 3mL, after vortex
It is placed at room temperature for 15min.After solution is thoroughly layered, the upper layer n-hexane layer of 1mL is drawn, another clean tube is transferred to, leads to N2
Dry up solution.It can find that test tube wall has more or less cholesterol trace being dried at this time, then the neighbour of 4mL is added into test tube
Phthalaldehyde solution, vortex 1min, is placed at room temperature for 10min, draws the concentrated sulfuric acid of 2mL, is slowly added to along test tube wall, is vortexed, room
Temperature places 10min, measures the light absorption value of cholesterol OD550nm.Finally, the cholesterol standard curve according to drafting calculates gallbladder
The content of sterol.
Degrading rate of cholesterol=(A-B)/A × 100%
A, B respectively indicates the cholesterol level after the cholesterol level and culture 48h after inoculation in culture solution in culture solution
Table 2 is the strains for degrading cholesterol ability of the present invention filtered out in test.
The lactic acid bacteria cholesterol degradation ability of the present invention of table 2
Embodiment 4: external probiotic properties research
One, acid resisting test
With reference to the method for Huang etc., to the quantity of BX95 adjustment bacterium to 1.0 × 109Cfu/mL, then 10% (v/v) connect
Kind amount is inoculated in the MRS fluid nutrient medium of pH 2.0, then carries out plate count to its viable count in 0h, 1h, 2 h and 3h
Method calculates survival rate, and draws the dynamic of its survival rate in 3h.The acid-fast ability that table 3 and Fig. 1 and Fig. 2 are BX95, as a result
It was found that BX95 bacterial strain, acid resistance is very good.
The acid-fast ability of 3 11 strains of lactic acid bacteria of table
Two, hydrophobicity tests
By bacterium solution, thalline were collected by centrifugation, after PBS (pH 6.5) buffer washes twice, adds PBS to be vortexed and mixes, bacteria suspension tune
The quantity of whole bacterium is to 1.0 × 109Its initial light absorption value A is measured at cfu/mL, 600nm0.The bacteria suspension of 3mL is drawn again, and 1mL is added
Dimethylbenzene, be stored at room temperature culture 10min, vortex mix well 2min, be stored at room temperature 15min, after its layering completely after, carefully
Lower layer's water phase is drawn, clean tube is transferred to.Using buffer as blank control, it is measured in the final light absorption value A of 600nm, note
Record calculates hydrophobic rate.
Lactobacillus cell surface hydrophobicity rate H%=(A0-A)/A0× 100%
A0It is respectively indicated with A and mixes the light absorption value that front and back measures at 600nm with dimethylbenzene
Three, simulation gastro-intestinal Fluid test
Thalline were collected by centrifugation, and sterile saline is vortexed after washing 2-3 times, and thallus is made to suspend.Bacteria suspension 1mL is drawn in
In 9mL simulated gastric fluid (pH 2.5) through degerming, it is vortexed after mixing and samples 0.5mL immediately, 10 times of sterile saline dilutions
Afterwards, the dilution 0.1mL of appropriate dilution is chosen on MRS solid plate, and coating is uniform.3 repetitions are set.Remaining bacterium
Liquid is in 37 DEG C of constant incubator stationary culture 3h.After digesting 3h, 0.5mL sample is drawn, dilution applies plate, 37 DEG C of constant incubators
It is counted after Anaerobic culturel 48h.
The bacteria-containing simulated gastric fluid that 1mL has handled 3h is drawn, is inoculated in the simulated intestinal fluid of 9mL degerming, 37 DEG C
Anaerobism constant temperature stationary culture, then respectively 0,3,6,9,20 and for 24 hours sample 0.5mL sample, apply plate, 37 DEG C of constant temperature incubations
Case Anaerobic culturel 48h is counted.Test three repetitions of setting.Colony counting method calculates viable count, counts the flat of 30-300 bacterium colony
Plate.And calculate its survival rate (%).
Survival rate (%)=(Lg N/Lg N0) × 100%
N0The viable count in 0h and digestive juice after digestion certain hour is respectively indicated with N
Four, lactic bacteria strain bile tolerance is tested
The quantity of lactic acid bacteria adjustment bacterium in 2-3 generation will be activated to 1.0 × 109Cfu/mL, then the inoculum concentration of 1% (v/v) connect
Plant setting three in MRS+Thio culture solution (control group) and in the MRS+Thio culture solution (test group) containing 0.3% bovine bile
A repetition.It is mixed after inoculation.Sampling 5mL measures the light absorption value of 620 nm immediately, and record, this is initial value.Bacterium solution is put later
37 DEG C of constant incubator stationary cultures are placed in, every two hours, the light absorption value of the OD620nm of two kinds of culture solutions is measured by sampling.Meter
The time that light absorption value changes 0.3 unit is calculated, the growth curve of bacterial strain is drawn.The calculating of retardation time (LT) is according to lactic acid bacteria
The light absorption value for growing into OD620nm changes the time used when 0.3 unit.LT is smaller.Show the stronger (stone of the ability of bile tolerance
It is super, 2013).
The hydrophobic ability of 4 invention bacterial strain BX95 of table and simulation alimentary canal tolerance
1 lactic acid bacteria is survived in pH2.5 simulated gastric fluid after 3h, then the survival rate after digesting for 24 hours in simulated intestinal fluid.
2 in absorbance OD620nm, and bacterial strain is raw in two kinds of culture mediums of MRS-THIO and MRS-THIO (containing 0.3% cholate)
It is long to increase the time difference required for 0.3 unit.
Table 4 shows that bacterial strain of the present invention has preferable gastrointestinal tract tolerance and hydrophobic ability, in addition to bile tolerance ability
It is poor, and hydrophobicity increases the chance for being attached to intestinal walls, meets the primary condition as probiotics.
Five, lactic acid bacteria bacteriostatic test
Bacteriostatic test (Papamaloni et al., 2003) is carried out using agar diffusion method.The step of having, is changed.
The preparation of lactobacillus-fermented supernatant: the quantity of lactic acid bacteria adjustment bacterium in 2-3 generation will be activated to 1.0 × 108cfu/
ML, 2% (v/v) are inoculated into MRS fluid nutrient medium, and 37 DEG C of stationary cultures are for 24 hours.10000rpm/min, 4min take supernatant.Nothing
0.22 μm of the membrane filtration degerming of bacterium.This is lactobacillus-fermented supernatant.It is spare to be placed in 4 DEG C of refrigerators.
Nutrient medium after sterilizing is cooled to 50 DEG C or so, is respectively connected to the pathogenic bacteria cultivated, bacterium number is adjusted, makes
Its final concentration of 1.0 × 106Cfu/mL, inverted plate after mixing.After culture medium cooled and solidified, existed with the aseptic card punch of 8mm
It is punched on plate, two holes of each plate, is separately added into the lactobacillus-fermented supernatant of 0.1 mL and the MRS culture without bacterium solution
Liquid is as control.It marks.After 4 DEG C of diffusion 4-5h for 24 hours in 37 DEG C of constant incubator cultures.Vernier caliper measures inhibition zone
Diameter, deduct the aperture 8mm size.
Table 5 shows the fermentation liquid for 24 hours of invention bacterial strain BX95 to staphylococcus aureus, listeria spp, Escherichia coli
Stronger rejection ability is all showed with salmonella typhimurium, and there is relatively broad bacteriostasis.
The bacteriostasis of 5 invention bacterial strain BX95 of table
Embodiment 5:SD rat in vivoassay
One, experimental animal and grouping
Experiment uses 5 week old male SD rat 30, and between 185.8-287.2g, average value is weight range
240.38g.Every is fed respectively in the mouse cage being individually isolated, and room temperature maintains 23 ± 2 DEG C, and relative humidity is 55 ± 5%, daily
The periodically 12 hours dark of illumination in 12 hours.In the meantime, the free feeding feed of rat and drinking-water.SD rat was in first week laundering period
The feed of institute's feeding is pulled together feed corporation,Ltd purchased from Beijing Australia, section.It adapts to after a week, rat is randomly divided into three groups, every group 10
Only, group is as follows:
C: blank control group, this group of SD feeding rats normal diet (are pulled together feed corporation,Ltd) purchased from Beijing Australia, section
H group: diet group high in fat, this group of feeding rats high cholesterol diet (are pulled together feed corporation,Ltd) purchased from Beijing Australia, section
HL group: daily ration high in fat+Lactobacillus helveticus BX95 group, this group of SD feeding rats high cholesterol diet, and fill daily
Stomach 1mL lactic acid bacterial liquid, bacterium number is 1 × 109cfu/mL。
Two, content is measured
Rat body weight is recorded weekly, acquires fresh excreta, it is aqueous to calculate excrement after 105 DEG C of oven drying 48h with aluminium box
Amount, is as a result shown in Fig. 3.The fecal water for feeding the later rat of daily ration high in fat as the result is shown can be decreased obviously, but feeding Switzerland is newborn
The fecal water of bacillus BX95 group rat can be higher than the fecal water of high in fat group of rat.
After experiment carries out 8 weeks, by etherization after Rat Fast 12h, Culling heart blood 2mL is put into 4 DEG C of refrigerators.Furthermore by liver
Dirty, spleen, kidney taking-up normal saline flushing are simultaneously weighed after being dried with filter paper, are calculated organ index, be the results are shown in Table 6.As a result
Show that feeding Lactobacillus helveticus BX95 does not have obvious toxic-side effects to rat, and it is heavy in liver organization to reduce fat
Product.
Each organ index of 6 rat of table compares
Note: a b c indicates the existing significance of difference between corresponding index, indicates significant difference (P with column data Superscript letters
< 0.05), the identical expression difference of letter is not significant (P > 0.05)
It finally cuts liver organization block (1cm × 1cm) and is put into fixation in 10% formalin.
Three, blood serum sample preparation and each index analysis of cholesterol
Collected blood sample places 4 DEG C of refrigerator overnights, and low temperature low-speed centrifugal takes upper layer weak yellow liquid, freezes immediately
In -20 DEG C of refrigerators, until measuring each index of cholesterol.
Total cholesterol level, content of triglyceride, low-density lipoprotein content, hdl concentration in blood serum sample
Four indexs, are measured using the conventional kit of commercially available corresponding index, as a result see Fig. 4,5,6,7.The result shows that Switzerland is newborn
Total cholesterol in rat blood serum, triglycerides, low-density lipoprotein content can be significantly reduced in bacillus BX95, but to high density rouge
Protein content is not substantially change.Four, pathology of hepar is sliced
The liver organization that fixes carry out paraffin embedding, slice, fixation, HE dyeing and etc. obtain pathology of hepar
Slice is sliced under the microscope in 100 times of mirror optical microphotographs, as a result sees Fig. 8,9,10.The result shows that Lactobacillus helveticus BX95 can be with
Effectively reduce the fatty infiltration of liver cell.
Bacterial strain Lactobacillus helveticus BX95 provided by the invention tests prove that, norcholesterol ability with higher.It should
Bacterial strain meets the primary condition as probiotics, the ability with stronger inhibition common pathogen growth, just to maintenance enteron aisle
Normal microbial flora balance tool has certain effect, and is able to maintain certain survival rate and passes through digestive tract environment.The bacterial strain is preferably dredged
Outlet capacity increases it and is attached to intestinal walls and plays the chance of prebiotic effect.
SEQUENCE LISTING
<110>Gansu Pu Nuobeikang biotechnology Co., Ltd
<120>one plants have the Lactobacillus helveticus for reducing cholesterol ability
<130> 6
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1314
<212> DNA
<213>Lactobacillus helveticus (Lactobacillus helveticus)
<400> 1
acgggcggtg tgtacaaggc ccgggaacgt attcaccgcg gcgttctgat ccgcgattac 60
tagcgattcc agcttcgtgc agtcgagttg cagactgcag tccgaactga gaacagcttt 120
cagagattcg cttgccttcg caggctcgct tctcgttgta ctgcccattg tagcacgtgt 180
gtagcccagg tcataagggg catgatgact tgacgtcatc cccaccttcc tccggtttgt 240
caccggcagt ctcattagag tgcccaactt aatgctggca actaataata agggttgcgc 300
tcgttgcggg acttaaccca acatctcacg acacgagctg acgacagcca tgcaccacct 360
gtcttagcgt ccccgaaggg aactcctaat ctcttaggat ggcactagat gtcaagacct 420
ggtaaggttc ttcgcgttgc ttcgaattaa accacatgct ccaccgcttg tgcgggcccc 480
cgtcaattcc tttgagtttc aaccttgcgg tcgtactccc caggcggagt gcttaatgcg 540
ttagctgcag cactgagagg cggaaacctc ccaacactta gcactcatcg tttacggcat 600
ggactaccag ggtatctaat cctgttcgct acccatgctt tcgagcctca gcgtcagttg 660
cagaccagag agccgccttc gccactggtg ttcttccata tatctacgca ttccaccgct 720
acacatggag ttccactctc ctcttctgca ctcaagaaaa acagtttccg atgcaattcc 780
tcggttaagc cgagggcttt cacatcagac ttattcttcc gcctgcgctc gctttacgcc 840
caataaatcc ggacaacgct tgccacctac gtattaccgc ggctgctggc acgtagttag 900
ccgtgacttt ctggttgatt accgtcaaat aaaggccagt tactacctct atccttcttc 960
accaacaaca gagctttacg atccgaaaac cttcttcact cacgcggcgt tgctccatca 1020
gacttgcgtc cattgtggaa gattccctac tgctgcctcc cgtaggagtt tgggccgtgt 1080
ctcagtccca atgtggccgt tcagtctctc aactcggcta tgcatcattg ccttggtaag 1140
ccgttacctt accaactagc taatgcaccg cggggccatc ccatagcgac agcttacgcc 1200
gccttttata agctgatcat gcgatctgct ttattatccg gtattagcac ctgtttccaa 1260
gtggtatcct agactatggg gcaggttccc cacgtgttac tcacccatcc gccg 1314
Claims (6)
1. one plant has the Lactobacillus helveticus (Lactobacillus helveticus) for reducing cholesterol ability, feature exists
In the deposit number of the Lactobacillus helveticus is CGMCC15778.
2. the one plant as described in claim 1 screening technique with the Lactobacillus helveticus for reducing cholesterol ability, feature exist
In separation has the lactic acid bacteria strains of norcholesterol from Qinghai-Tibet traditional zymotic yak yoghourt.
3. the one plant as claimed in claim 2 screening technique with the Lactobacillus helveticus for reducing cholesterol ability, it is characterised in that
It screens in the steps below:
(1) isolate and purify: the microorganism in Qinghai-Tibet traditional zymotic yak yoghourt passes through tetra- kinds of differences of MRS, M17, KFS and SL
Culture medium culture, select feature bacterial strain and purify, obtain pure isolated bacterial strain；
(2) bacterial strain is identified: in conjunction with hydrogen peroxide feminine gender, Gram-positive, thalli morphology and microscopy separating, selecting like cream
Sour bacteria strain carries out 16S rDNA sequencing, carries out Blast comparison in NCBI, identify and save 720 strains of lactic acid bacteria bacterial strains；
(3) preliminary screening: solid by the drop gallbladder of the lactic acid bacteria strains identified in o-phthalaldehyde colorimetric method for determining step (2)
Alcohol ability filters out the ability that bacterial strain BX95 of the present invention has high norcholesterol, is accredited as Lactobacillus helveticus in (2)
(Lactobacillus helveticus)；
(4) the 16S rRNA gene order of bacterial strain BX95 is registered in Genbank, obtains Genbank database bacterial strain
The sequence number of BX95: MF093231.
4. the one plant as described in claim 1 Lactobacillus helveticus with norcholesterol ability, it is characterised in that by the Switzerland
The 16S rRNA gene order of lactobacillus strain BX95 is registered in Genbank, obtains Genbank database bacterial strain BX95's
Sequence number: MF093231.
5. the Lactobacillus helveticus with high norcholesterol is preparing the application in fermented dairy product as described in claim 1.
6. the Lactobacillus helveticus with high norcholesterol is preparing answering in health food and drug as described in claim 1
With.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113430135A (en) * | 2021-07-01 | 2021-09-24 | 杭州娃哈哈科技有限公司 | Lactobacillus helveticus strain with depression improving effect and application thereof |
| CN115466705A (en) * | 2022-11-14 | 2022-12-13 | 中国食品发酵工业研究院有限公司 | Lactobacillus helveticus fermentation product with high biological preservative property and preparation method and application thereof |
| CN116064286A (en) * | 2022-08-19 | 2023-05-05 | 浙江大学 | Lactobacillus helveticus ZJUIDS11 for improving nonalcoholic liver disease and application thereof |
| CN116574611A (en) * | 2023-04-26 | 2023-08-11 | 甘肃普诺贝康生物科技有限责任公司 | Freeze-drying protective agent for lactobacillus buchneri and method for preparing lactobacillus buchneri freeze-drying powder by using freeze-drying protective agent |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU94013100A (en) * | 1994-04-13 | 1997-04-10 | Московский научно-исследовательский институт эпидемиологии и микробиологии им.Г.Н.Габричевского | Strain of lactobacillus helveticus tiii exhibiting capability to decreased blood serum cholesterol level |
| CN104073455A (en) * | 2014-06-18 | 2014-10-01 | 兰州大学 | Lactobacillus plantarum capable of lowering cholesterol |
| BG66608B1 (en) * | 2011-05-04 | 2017-10-16 | "Ел Би Булгарикум" ЕАД | Polybacterial probiotic preparation |
-
2018
- 2018-08-17 CN CN201810941393.4A patent/CN109456909A/en not_active Withdrawn
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU94013100A (en) * | 1994-04-13 | 1997-04-10 | Московский научно-исследовательский институт эпидемиологии и микробиологии им.Г.Н.Габричевского | Strain of lactobacillus helveticus tiii exhibiting capability to decreased blood serum cholesterol level |
| BG66608B1 (en) * | 2011-05-04 | 2017-10-16 | "Ел Би Булгарикум" ЕАД | Polybacterial probiotic preparation |
| CN104073455A (en) * | 2014-06-18 | 2014-10-01 | 兰州大学 | Lactobacillus plantarum capable of lowering cholesterol |
Non-Patent Citations (2)
| Title |
|---|
| 张娟: "青藏高原传统发酵牦牛酸奶中乳酸菌的降胆固醇及体外益生特性研究", 《中国优秀硕士学位论文全文数据库 工程科技I辑》 * |
| 杨丽杰等: "乳酸菌降胆固醇机制的研究进展", 《食品与发酵工业》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113430135A (en) * | 2021-07-01 | 2021-09-24 | 杭州娃哈哈科技有限公司 | Lactobacillus helveticus strain with depression improving effect and application thereof |
| CN116064286A (en) * | 2022-08-19 | 2023-05-05 | 浙江大学 | Lactobacillus helveticus ZJUIDS11 for improving nonalcoholic liver disease and application thereof |
| CN116064286B (en) * | 2022-08-19 | 2023-08-11 | 浙江大学 | Lactobacillus helveticus ZJUIDS11 for improving nonalcoholic liver disease and application thereof |
| CN115466705A (en) * | 2022-11-14 | 2022-12-13 | 中国食品发酵工业研究院有限公司 | Lactobacillus helveticus fermentation product with high biological preservative property and preparation method and application thereof |
| CN115466705B (en) * | 2022-11-14 | 2023-08-18 | 中国食品发酵工业研究院有限公司 | Lactobacillus helveticus fermentation product with high biological preservative property, and preparation method and application thereof |
| CN116574611A (en) * | 2023-04-26 | 2023-08-11 | 甘肃普诺贝康生物科技有限责任公司 | Freeze-drying protective agent for lactobacillus buchneri and method for preparing lactobacillus buchneri freeze-drying powder by using freeze-drying protective agent |
| CN116574611B (en) * | 2023-04-26 | 2023-10-13 | 甘肃普诺贝康生物科技有限责任公司 | Freeze-drying protective agent for lactobacillus buchneri and method for preparing lactobacillus buchneri freeze-drying powder by using freeze-drying protective agent |
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