CN101328469B - Streptococcus thermophilus grx02 having alcoholic liver damage protection function and use thereof - Google Patents
Streptococcus thermophilus grx02 having alcoholic liver damage protection function and use thereof Download PDFInfo
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Abstract
The invention provides a streptococcus thermophilus grx02 capable of protecting alcoholic liver injury and a preparation method as well as an application thereof, relating to the biotechnological filed, in particular to an isolated strain of a functional lactic acid bacterium and an application thereof. The separation process is performed by adopting a culture medium in a Xinjiang nationality traditional dairy product to obtain the lactic acid bacterium; the lactic acid bacterium with the antioxidation activity, anti-pathogen property and the ability to damage the endotoxin ability is selected; the lactic acid bacterium capable of protecting the alcoholic liver injury is selected in the selected lactic acid bacterium by an animal experiment comparison, and the finally selected lactic acidbacterium is the streptococcus thermophilus grx02. The functional lactic acid bacterium has functional property in the liver injury protecting aspect. If the streptococcus thermophilus grx02 is developed into a health care food and is taken for a long time, the liver injury can be prevented, relieved and treated auxiliarily.
Description
Technical field
The present invention relates to biological technical field, particularly the strain isolated of functional lactobacillus and application.
Technical background
To the protection research of the liver injury that caused by alcohol, the Western European countries stress the treatment in alcohol acute poisoning or alcohol hepatopathy, based on passive methods of treatment; Country in Southeast Asia comprises China, and most of research starting point is a purpose to relieve the effect of alcohol, to sober up, and has ignored prevention, regulates this important step.And entering intravital medicine great majority will be through the conversion of liver.Liver problem sufferer's hypofunction of liver, many medicines enter in the body, certainly will increase burden of liver.These materials are metabolism further liver injury again in liver, forms the accumulated damage process, can further damage liver on the contrary.
Milk-acid bacteria has the adjusting physiologically active, and preventing disease takes place and the high characteristics of security.The alcoholic liver injury provide protection of milk-acid bacteria is that it has anti-oxidant activity, and prevention liver lipid peroxidation is effectively removed harmful free radical that liver injury produces; Have the growth of harmful bacterium such as Chinese People's Anti-Japanese Military and Political College enterobacteria, Salmonellas, keep the intestinal microflora balance, reduce the intracellular toxin generation and absorb the generation of bacterial translocation and reduction endotoxemia when effectively preventing liver injury.
The kind of the power milk-acid bacteria employed with it of the liver injury protection effect of cultured milk prod is closely related.Filtering out the bacterial classification that is fit to the production liver injury protection from numerous lactobacillus inoculations has great importance.Be developed to protective foods as the milk-acid bacteria that will have the liver injury protection effect, take for a long time can prevent, alleviation and assisting therapy liver injury, this mode will be accepted by people.In food today to the development of natural type and functional type, producing this product of functional yogurt is one of direction of lactobacillus product development, and alcoholic liver damage protection function milk-acid bacteria Products Development has than wide prospect, has had much market potential.
Summary of the invention
The object of the invention is to invent a kind of milk-acid bacteria with alcoholic liver injury provide protection---streptococcus thermophilus grx 02 in order to overcome aforementioned and other shortcoming.
The present invention is preserved on May 28th, 2008, is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center, the address: Datun Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica.Preserving number is CGMCC No:2525.
Through advance copy invention is a kind of functional lactobacillus, has functional performance especially aspect liver injury protection, as streptococcus thermophilus grx 02 of the present invention is developed to protective foods, take for a long time can prevent, alleviation and assisting therapy liver injury.
Another object of the present invention is to invent the preparation method of above-mentioned streptococcus thermophilus grx 02:
May further comprise the steps:
1) adopts substratum from the national tradition milk-product of Xinjiang, to separate and obtain milk-acid bacteria;
2) filter out have anti-oxidant activity, anti-pathogenic bacterium characteristic and the milk-acid bacteria of destroying the intracellular toxin ability;
3) by experimentation on animals relatively from through step 2) filter out the milk-acid bacteria that alcoholic liver injury is had defencive function the milk-acid bacteria that filters out, be streptococcus thermophilus grx 02.
The milk-acid bacteria that the above method of process obtains determines that by the evaluation that the French Mei Aili API of company reagents series bar and 16SrDNA sequencing are carried out this bacterial strain is thermophilus streptococcus (Streptococcusthermopilus), identifies that accuracy rate is all above 99%.
The 3rd purpose of the present invention is the application of the described streptococcus thermophilus grx 02 of invention:
Can be used for preparing food with alcoholic liver damage protection function.
Also can be used for preparing fermented-milk with alcoholic liver damage protection function.
The various products of strain fermentation provided by the invention can play the effect of protection alcoholic liver injury; the milk of this strain fermentation; the various fermented milk prods through lactobacillus-fermented that obtain, lactobacillus drink product etc. have all obtained to have the characteristic of protection alcoholic liver injury in various degree.
Utilize strain fermentation skimmed milk of the present invention or other food raw material,, have preferably stability and, still be used to prepare the better selection of pharmaceutical composition with alcoholic liver damage protection function than the long products shelf-lives through processing treatment such as super-dries.Take for a long time can prevent, alleviation and assisting therapy liver injury, be particularly suitable for long-term heavy drinking personnel, with without any side effects.
Description of drawings
Fig. 1 is streptococcus thermophilus grx 02 gramstaining microscopic examination figure.
Fig. 2 is the fluorescence pattern of streptococcus thermophilus grx 02 DNA.
Fig. 3 is the fluorescence pattern of streptococcus thermophilus grx 02 16S rDNA-PCR product.
Fig. 4 is the HE dyeing picture (200 times) of the liver slice of control group mice.
Fig. 5 is the HE dyeing picture (200 times) of the liver slice of model group mouse.
Fig. 6 is the HE dyeing picture (200 times) of the liver slice of medicine group mouse.
Fig. 7 is the HE dyeing picture (200 times) of the liver slice of milk-acid bacteria contrast C8M group mouse.
Fig. 8 is the HE dyeing picture (200 times) of the liver slice of streptococcus thermophilus grx 02 group mouse.
Embodiment
The present inventor attempts to obtain new strains of streptococcus thermophilus.This bacterial strain cow's milk that can ferment shows the functional performance with liver injury protection---anti-oxidant activity, anti-pathogenic bacterium characteristic, destroy the intracellular toxin ability.
Invention is by separating the lactic bacterium strains that utilizes research anti-oxidant activity, anti-pathogenic bacterium characteristic in a large amount of milk-acid bacterias that obtain from the tradition milk-product of Xinjiang.Again by ne ar, physiology and cultural characteristic, and carry out Analysis and Identification by the French Mei Aili API of company reagents series bar, the bacterial strain 16SrDNA Gene Partial sequence of birdsing of the same feather flock together, the result has confirmed that bacterial strain of the present invention is a streptococcus thermophilus grx 02.
1. sample source
Bacterium source of the present invention is in Xinjiang of China national tradition milk-product-koumiss.Koumiss (Koumiss) is to be raw material with fresh mare's milk, the low ethanol content milky-drinks of acidity that forms through the common spontaneous fermentation of microorganisms such as milk-acid bacteria and yeast.
2. sample collecting
To put into previously prepd buffered soln behind the sample collecting, refrigeration separated sample within 2 days.
3. milk-acid bacteria separates
With Xinjiang national tradition milk-product koumiss sample adopt the PTYG culture in 37 ℃ cultivate 24 hours after.This culture is coated on the PTYG agar plate, cultivated 3 days at 37 ℃ then, picking list bacterium colony, and be further purified, the bacterial detection feature, gramstaining as mentioned below is as shown in Figure 1.
After the bacterial strain that separate to obtain cultivated with the culture that contains 11% skimming milk, add protective material, freeze-drying, be kept at-18 ℃, be used for the test of various bacterial detection features then.
4. the evaluation of bacterial strain
An enterprising step carries out that API identifies and the 16SrDNA sequencing identifies that above-mentioned bacterium is a streptococcus thermophilus grx 02 on Physiology and biochemistry basis.
(1) use the French Mei Aili API of company series indentifying substance bar to carry out the sugar-fermenting analysis.
At 37 ℃, under anaerobic streptococcus thermophilus grx 02 was cultivated 48 hours on the MRS agar plate, with the suspension culture base gained culture is suspended then, the turbidity of suspension is identical with McFarland's 2.Then, this culturing cell is inoculated in the API 50CHL reagent strip, layer covers the aseptic paraffin of one deck thereon, so that detect the fermentation capacity to 49 kinds of sugar.
The sugar-fermenting ability of having used the programanalysis of API identification software.The result shows, the sugar-fermenting ability of this bacterial strain and thermophilus streptococcus similar (table 1), and coincidence rate is 96.3%.
(2) bacterial strain 16SrDNA Sequence Identification.
Adopt bacterial genomes DNA extraction agent box in a small amount, operate according to explanation, institute's streptococcus thermophilus grx 02 genome dna electrophoresis band of carrying is clear, illustrates that the genomic dna integrity better can be used for PCR equimolecular biologic operation (as shown in Figure 2).
As template, adopt 50 μ l reaction systems to carry out pcr amplification with genomic dna, template DNA 5 μ l (approximately 100ng) wherein, 2.5mM dNTP 4 μ l, 10 * buffer, 5 μ l, each 1.5 μ l of 10pmol upstream and downstream primer, Taq enzyme 1.5U, Mg2+3 μ l is with ddH
2O mends to 50 μ l.The PCR loop parameter is: 94 ℃ of pre-sex change 5min; 94 ℃ of sex change 1min, 64 ℃ of annealing 45s, 72 ℃ are extended 1min, circulate 35 times; 72 ℃ are extended 8min.Draw PCR product 5 μ l and mix with 1 μ l Loading buffer, use 1.0% agarose gel electrophoresis, voltage is 80V.Electrophoresis finishes the back with ethidium bromide (EB) dyeing 20-30min, takes pictures.As seen the band of about 1500bp is target stripe (as shown in Figure 3).
Examining order is finished by Shanghai Sangon Biological Engineering Technology And Service Co., Ltd, sequencing result: the 16SrDNA sequence length of streptococcus thermophilus grx 02 is 1456bp.With reported sequence comparison among the Genbank, bacterial strain of the present invention is proved up to 99.65% with homology and belongs to thermophilus streptococcus.
API identifies consistent with 16SrDNA sequencing result, and bacterial strain of the present invention is thermophilus streptococcus (S.thermophilus) grx02.
5. acid-resistant property
In broth culture, mixed with 11% degreasing milk medium by 1: 1, with the HCl adjusting pH value of 10mol/L, the pH value is adjusted into 1.0,1.5,2.0,2.5,3.0,3.5 respectively.Inoculate 3% streptococcus thermophilus grx 02 nutrient solution then, 37 ℃ of anaerobism were cultivated 2 days.Found that streptococcus thermophilus grx 02 37 ℃ of cultivation 48h number of viable when pH value 2.0 are 10
4-5Cfu/mL.
The acid-resistant property of table 2 streptococcus thermophilus grx 02 of the present invention
6. bile tolerance characteristic
Specifically be, in order to study the anti-bile ability of bacterial strain, in 100mL, 11% skimming milk, add the pig cholate, make biliary concentration be respectively 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5% and 4.0%, then with 1mL (10
9About cfu/mL) the streptococcus thermophilus grx 02 culture be inoculated in these skimming milks, respectively anaerobism was cultivated 2 days.Use the MRS broth culture that is kept at 4 ℃ with 10 times of stepwise dilution nutrient solutions then, afterwards diluent is applied to the flat anaerobism of pulling and cultivates, detect viable count.
The anti-bile characteristic of table 3 streptococcus thermophilus grx 02 of the present invention
Drug concentration in bile be respectively in 1.0%～2.5% the skimming milk 37 ℃ on bacterium when cultivating 2 days number of viable all 10
5-8In the scope of cfu/mL; Drug concentration in bile is that number of bacteria is 10 in 3.0% and 3.5% the skimming milk
3-5Cfu/mL; Drug concentration in bile is to cultivate amount of viable bacteria through 2 days in 4.0% the skimming milk still can surpass 10
2Cfu/mL proves that bacterial strain of the present invention has resistance to bile.
7. anti-oxidation characteristics
(1) SOD activity
Specifically be, be inoculated in 15mL through three activatory streptococcus thermophilus grx 02s by 3% inoculum size, in 11% skimming milk, cultivate at 37 ℃, fermented liquid is at 4 ℃, centrifugal 20min under the 3500rpm condition.Get supernatant liquor, behind the accent pH to 6.0, adopt pyrogallol autoxidation method to measure the SOD activity.Be divided into from 26 strains of lactic acid bacteria that obtain from Xinjiang, the SOD activity accounts for 15.38% greater than the bacterial strain number of 25U/mL, bacterial strain number less than 20U/mL accounts for 38.46%, and the SOD activity of streptococcus thermophilus grx 02 is apparently higher than other bacterial strain, and its active quantities can reach 28.77U/mL.
(2) remove DPPH free radical ability
Specifically be, be inoculated in 15mL through three activatory streptococcus thermophilus grx 02s by 3% inoculum size, in 11% skimming milk, cultivate at 37 ℃, bacterial strain fermentation liquor is at 4 ℃, under the 3500rpm condition behind the centrifugal 20min, get supernatant liquor, be diluted to different percentage concentrations by volume with 95% ethanol, with the strain fermentation supernatant liquor and the equal-volume concentration of different weaker concns is that 30mg/L DPPH solution adds in the same tool plug test tube, shake up, measure its absorbancy with 95% ethanol as reference behind the 30min, calculate clearance rate.Fermented liquid supernatant is removed the DPPH ability all along with its removing ability of rising of concentration strengthens, and fermented liquid concentration and DPPH clearance rate are linear.Be divided into from 26 strains of lactic acid bacteria that obtain from Xinjiang, when removing 50%DPPH, the fermented liquid concentration of required consumption accounts for 11.54% less than 3.40% bacterial strain number, and the fermented liquid concentration of required consumption accounts for 61.54% greater than 3.60% bacterial strain number.The removing DPPH ability of streptococcus thermophilus grx 02 is apparently higher than other bacterial strain, and the grx02 fermented liquid concentration of required consumption is 3.34%.
(3) Total antioxidant capacity
Specifically be, be inoculated in 15mL through three activatory streptococcus thermophilus grx 02s by 3% inoculum size, in 11% skimming milk, cultivate at 37 ℃, bacterial strain fermentation liquor behind the centrifugal 20min, is got supernatant liquor at 4 ℃ under the 3500rpm condition, transfers pH 6.0.Measure Total antioxidant capacity (T-AOC) by test kit specification sheets (biotechnology company limited is built up in Nanjing).Be divided into from 26 strains of lactic acid bacteria that obtain from Xinjiang, Total antioxidant capacity accounts for 15.38% greater than the bacterial strain number of 15U/mL, bacterial strain number less than 10U/mL accounts for 69.23%, and the streptococcus thermophilus grx 02 Total antioxidant capacity is apparently higher than other bacterial strain, and its antioxidant capacity can reach 17.35U/mL.
In sum, streptococcus thermophilus grx 02 of the present invention has higher anti-oxidant activity.
8. to the rejection characteristics of pathogenic bacterium
(1) intestinal bacteria is suppressed ability
Adopt individual layer agar plate diffusion process.Dust Xi Shi intestinal bacteria are inoculated in the liquid LB substratum, behind 37 ℃ of shaking culture 4h, colony counting method meter viable count, 4 ℃ of refrigerations, standby.Control the intestinal bacteria viable count 10 by dilution
6Cfu/mL, this concentration is easily caused a disease.Draw Escherichia coli bacteria liquid 0.1mL respectively on solid LB culture medium flat plate, with aseptic paint daubs coating evenly, logical sterile air in Bechtop, place 1h and treat liquid-solid back, the surface punching that fixes on flat board of bacterium on surface, the diameter 10mm in hole, hole depth 3mm is with bacterial strain sample liquid 0.2mL (10
8Cfu/mL) in the filling orifice, cultivate 8h observations, mensuration antibacterial circle diameter in 37 ℃.Each dull and stereotyped three hole, it is parallel to make two flat boards, is used for analyzing.Be divided into from 26 strains of lactic acid bacteria that obtain from Xinjiang, bacteriostatic diameter accounts for 26.92% greater than the bacterial strain number of 22mm, accounts for 46.15% less than the bacterial strain number of 20mm, and it is relatively good that streptococcus thermophilus grx 02 suppresses the intestinal bacteria ability, and bacteriostatic diameter is about 23mm.
(2) Salmonellas is suppressed ability
Salmonella typhi is inoculated in the liquid LB substratum, behind 37 ℃ of cultivation 6h, colony counting method meter viable count, 4 ℃ of refrigerations, standby.Operation steps is the same.Be divided into from 26 strains of lactic acid bacteria that obtain from Xinjiang, bacteriostatic diameter accounts for 30.77% greater than the bacterial strain number of 24mm, accounts for 46.15% less than the bacterial strain number of 22mm, and it is relatively good that streptococcus thermophilus grx 02 suppresses the Salmonellas ability, and bacteriostatic diameter is about 26mm.
In sum, streptococcus thermophilus grx 02 of the present invention has stronger anti-pathogenic bacterium ability.
9. when cultivating altogether, destroy the intracellular toxin characteristic
With 0.2mL concentration is intracellular toxin and the 0.1mL (10 of 10eu/mL
8Cfu/mL) bacterium liquid is connected in the MRS nutrient solution together, make the intracellular toxin final concentration at 1eu/mL, intracellular toxin and the milk-acid bacteria of 1eu/mL are cultivated altogether at 37 ℃, when 6h, 12h, 24h, utilize tachypleus amebocyte lysate box (Shanghai City medical science assay office) to measure endotoxin content, observe the different incubation times of milk-acid bacteria and destroy the intracellular toxin ability.Be total under the culture condition with intracellular toxin, be divided into from 26 strains of lactic acid bacteria that obtain from Xinjiang, with respect to positive control OD value about 0.86, bacterial strain destroys intracellular toxin to OD value and drops to 0.30 bacterial strain number and account for 19.23%, streptococcus thermophilus grx 02 can obviously destroy intracellular toxin, along with the prolongation of incubation time destroys endotoxic ability in various degree raising is arranged, the OD value can drop to 0.19.Illustrate that streptococcus thermophilus grx 02 of the present invention has stronger destruction intracellular toxin ability.
10. protect the alcoholic hepatic injury characteristic
The milk product through this strain fermentation that obtains through the experiment of chmice acute alcoholic liver injury, has obtained the characteristic of protection liver injury.
90 of Kunming mouses, male and female half and half, body weight 22～24g provides available from Tang Shan Green Dragon mountain, Jiangning, Nanjing Experimental Animal Center; Conformity certification: SCXK (Soviet Union) 2002-0018.The shaft-like forage feed of standard recipe is freely intake.Mouse is raised and experiment is all carried out in that this school physician institute clean laboratory animal is indoor.
90 mouse are divided into 9 groups at random: blank group, model group, positive drug group, milk-acid bacteria control group C8M, streptococcus thermophilus grx 02 group, milk-acid bacteria component high dose group, middle dosage group, low dose group, 10 every group.Blank group: ad lib; Model group: adopt distilled water 10mL/kg continuous irrigation stomach 10d, with filling 50% alcohol 12mL/kg behind the last filling stomach 1h; The positive drug group: adopt DONGBAO GANTAI 10mL/kg (containing DONGBAO GANTAI 0.08g/mL) to irritate stomach, 10d and last are irritated 50% alcohol 12mL/kg after irritating stomach 1h continuously; Three groups of the high, medium and low dosage of milk-acid bacteria: adopt milk-acid bacteria 10.0mL/kg (108,107,106cfu/mL) continuous irrigation stomach 10d, irritate 50% alcohol 12mL/kg after irritating stomach 1h with last.Irritate after drinking, it is impaired enteron aisle to occur after the animal modeling, and two is just not normal, and hair is gradually withered matt, the One's spirits are drooping depressed performance that waits, and the above-mentioned general situation of each milk-acid bacteria group is obviously improved than model group, may to have an effect of protection enteron aisle relevant with milk-acid bacteria.Behind the fasting 12h, pluck eyeball and get blood, dissection is got liver and is measured each index.Result such as table 4, table 5.
Table 4 streptococcus thermophilus grx 02 to the influence of ALT, AST, TG, ALB, TP in the mice serum (n=10,
)
Annotate: compare with normal blank group: △ P＜0.05, △ △ P＜0.01; Compare with the alcohol model group:
*P＜0.05,
*P＜0.01
Table 5 streptococcus thermophilus grx 02 to the influence of MDA, GSH, SOD, ADH in the mouse liver (n=10,
)
Annotate: with table 4.
Can be found out by table 4,5: model group compares with each index of blank group, has significant difference (P＜0.05) the modelling success is described.Alcohol thorn is goaded into action and is made liver cell be subjected to a certain degree damage, and ALT, AST content obviously rise, and streptococcus thermophilus grx 02 has reduced hepatocellular damage (P＜0.05) effectively, and effect obviously is better than milk-acid bacteria control group C
8M.The total protein content of streptococcus thermophilus grx 02, albumin content are apparently higher than model group (P＜0.05), but dose relationship is indeterminate.The streptococcus thermophilus grx 02 antagonism reduction of activities of antioxidant enzymes due to the alcohol, suppressed the generation of free radical, the SOD activity is apparently higher than model group (P＜0.01), MDA content is starkly lower than model group (P＜0.01), reduced the exhaustion of liver GSH due to the alcohol, the GSH activity does not have significant difference apparently higher than model group (P＜0.01) and medicine group.And effect obviously is better than milk-acid bacteria control group C
8M.
Get respectively organize mouse liver lobus sinister same position estimate milk-acid bacteria in vivo protect the liver effect, the mouse pathological section is shown in Fig. 4 to 8: blank group central vein is arranged in liver lobule, the liver lobule clear in structure, the hepatic cords structural integrity is radial, and the liver cell kytoplasm is half-full, no reticulated structure, kernel is clear, and dikaryocyte is more, no inflammatory infiltration.Alcohol model group liver lobule boundary is unclear, and liver sinusoid is fuzzy to narrow down, liver cell kytoplasm muddiness, and the swelling of liver cell distortion has reticulated structure, and visibility is unintelligible, minority pyknosis, more inflammatory cell infiltration.The medicine group than model group for well, the mild inflammation cellular infiltration, pyknosis is slight.Milk-acid bacteria control group C
8The less inflammatory cell infiltration of M, minority swelling of liver cell, minor injury.The streptococcus thermophilus grx 02 group is basic near normal, and dikaryocyte is more.
11. bacterial strain preservation
Bacterial strain of the present invention is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center, address: Datun Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica on May 28th, 2008.Preserving number is CGMCC No:2525.
Use example
1, the making method that contains the yogurt of streptococcus thermophilus grx 02
Sweet milk is heated to more than 50 ℃, adds 6% white sugar and be stirred to dissolving fully, be preheating to 60～65 ℃, 20Mpa pressure homogeneous, 90 ℃ of left and right sides sterilizations 5-8 minute are cooled to 42～45 ℃, the inoculation streptococcus thermophilus grx 02.Inoculation quantity is 2～3%.40～42 ℃ ferment to the pH value be 4.2～4.5, the cooling, refrigeration, promptly make Solidify YoghurtJuzh.Fermentation ends refrigerates after jam fruit juice etc. is added in stirring, promptly makes to contain active streptococcus thermophilus grx 02 yogurt.
2, the making method that contains the streptococcus thermophilus grx 02 lactobacillus drink
Prepare yogurt by Application Example 1, with yogurt with mixture diluted 3-4 such as sterilization and refrigerative water, syrup, emulsion stabilizer doubly about, make the lactobacillus drink that contains active streptococcus thermophilus grx 02 capable of direct drinking.
3, contain the streptococcus thermophilus grx 02 manufacturing method of ice cream
Prepare yogurt by Application Example 1, the ice cream materials proportioning is as shown in table 5.
Table 5 1000kg ice-creams prescription
| Raw material | Weight (kg) |
| The streptococcus thermophilus grx 02 yogurt | 400 |
| White sugar | 75 |
| Whole milk powder | 62.5 |
| Yellow cream | 31.25 |
| Raw material | Weight (kg) |
| Oleum Cocois | 23 |
| Syrup | 26.25 |
| Emulsion stabilizer | 6 |
| Water | 276 |
Other mixing of materials that will be except that yogurt, be preheating to 60～65 ℃, adopt 20Mpa～30Mpa to carry out homogeneous,, be cooled to 40～50 ℃ then in 80 ℃/20s sterilization, mix with yogurt, adopt 20Mpa～30Mpa to carry out homogeneous, in the time of 2 ℃～4 ℃, digestion time 6h, through congealing after overvulcanization for the hard ice-creams, be ice milk without hardened.
4, the making method that contains the streptococcus thermophilus grx 02 powder food product
The streptococcus thermophilus grx 02 yogurt is refrigerated to-40 ℃～-60 ℃, drying, to water content be 3%-5%, be crushed to Powdered, powdery product can be added milk powder, oligose, maltodextrin, flavour agent etc. and make Powdered or the sheet bulk product, be used for daily edible.
More than the various products of strain fermentation provided by the invention can play the effect of protection alcoholic liver injury; the milk of this strain fermentation; the various fermented milk prods through lactobacillus-fermented that obtain, lactobacillus drink product etc. have all obtained to have the characteristic of protection alcoholic liver injury in various degree.
5, making method and the case that contains the streptococcus thermophilus grx 02 pharmaceutical composition prevented and treated example
Proportioning raw materials is as shown in table 6.
Table 6 feed proportioning table
| Raw material | Mass percent (%) |
| Skim-milk | 12.0 |
| Oligofructose | 2.0 |
| Yeast powder | 0.8 |
| Peptone | 0.7 |
| Pure water | 84.5 |
Proportionally skim-milk, lactose, yeast powder, peptone are mixed with pure water, be preheating to 60～65 ℃, 20Mpa pressure homogeneous, 90 ℃ of left and right sides sterilizations 20～30 minutes, be cooled to 42～43 ℃, inoculate 3% streptococcus thermophilus grx 02 starter, 42 ℃ ferment to the pH value be 4.1～4.5, centrifugal, lyophilize to water content less than 3%.Promptly be prepared into streptococcus thermophilus grx 02 lyophilize thing, in incapsulating after lyophilize thing and the maltodextrin balanced mix, promptly make the pharmaceutical composition that contains streptococcus thermophilus grx 02.
This product is taken for a long time and can be prevented, alleviation and assisting therapy liver injury, particularly long-term heavy drinking personnel.Take without any side effectsly for a long time, take 1-2 every day, contains number of viable of the present invention at least above 107 at every turn.
＜110〉Yangzhou University
＜120〉have alcoholic liver damage protection function streptococcus thermophilus grx 02 and preparation method thereof, use
<160>1
<210>1
<211>1456
<212>DNA
＜213〉milk-acid bacteria
<220>
＜223〉from the national tradition milk-product of Xinjiang, separate
<400>1
gggattggcg cgtgctatac atgcaagtag aacgctgaag agaggagctt gctcttcttg 60
gatgagttgc gaacgggtga gtaacgcgta ggtaacctgc cttgtagcgg gggataacta 120
ttggaaacga tagctaatac cgcataacaa tggatgacac atgtcattta tttgaaaggg 180
gcaattgttc cactacaaga tggacctgcg ttgtattagc tagtaggtga ggtaatggct 240
cacctaggcg acgatacata gccgacctga gagggtgatc ggccacactg ggactgagac 300
acggcccaga ctcctacggg aggcagcagt agggaatctt cggcaatggg ggcaaccctg 360
accgagcaac gccgcgtgag tgaagaaggt tttcggatcg taaagctctg ttgtaagtca 420
agaacgggtg tgagagtgga aagttcacac tgtgacggta gcttaccaga aagggacggc 480
taactacgtg ccagcagccg cggtaatacg taggtcccga gcgttgtccg gatttattgg 540
gcgtaaagcg agcgcaggcg gtttgataag tctgaagtta aaggctgtgg ctcaaccata 600
gttcgctttg gaaactgtca aacttgagtg cagaagggga gagtggaatt ccatgtgtag 660
cggtgaaatg cgtagatata tggaggaaca ccggtggcga aagcggctct ctggtctgta 720
actgacgctg aggctcgaaa gcgtggggag cgaacaggat tagataccct ggtagtccac 780
gccgtaaacg atgagtgcta ggtgttggat cctttccggg attcagtgcc gcagctaacg 840
cattaagcac tccgcctggg gagtacgacc gcaaggttga aactcaaagg aattgacggg 900
ggcccgcaca agcggtggag catgtggttt aattcgaagc aacgcgaaga accttaccag 960
gtcttgacat cccgatgcta tttctagaga tagaaagtta cttcggtaca tcggtgacag 1020
gtggtgcatg gttgtcgtca gctcgtgtcg tgagatgttg ggttaagtcc cgcaacgagc 1080
gcaaccccta ttgttagttg ccatcattca gttgggcact ctagcgagac tgccggtaat 1140
aaaccggagg aaggtgggga tgacgtcaaa tcatcatgcc ccttatgacc tgggctacac 1200
acgtgctaca atggttggta caacgagttg cgagtcggtg acggcgagct aatctcttaa 1260
agccaatctc agttcggatt gtaggctgca actcgcctac atgaagtcgg aatcgctagt 1320
aatcgcggat cagcacgccg cggtgaatac gttcccgggc cttgtacaca ccgcccgtca 1380
ccccacgaga gtttgtaaca cccgaagtcg gtgaggtaac cttttggagc cagccgccta 1440
aggtggacag atttgg
Claims (4)
1. have thermophilus streptococcus (Streptococcus thermopilus) grx02 of alcoholic liver damage protection function, it is characterized in that preserving number is CGMCC No:2525.
2. have the purposes of the streptococcus thermophilus grx 02 of alcoholic liver damage protection function according to claim 1, be used to prepare food with alcoholic liver damage protection function.
3. have the purposes of the streptococcus thermophilus grx 02 of alcoholic liver damage protection function according to claim 1, be used to prepare fermented-milk with alcoholic liver damage protection function.
4. have the purposes of the streptococcus thermophilus grx 02 of alcoholic liver damage protection function according to claim 1, be used to prepare pharmaceutical composition with alcoholic liver damage protection function.
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Families Citing this family (12)
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| CN101906391B (en) * | 2010-04-06 | 2011-08-24 | 扬州大学 | S.thermophilus grx90 for fermenting soya-bean milk products and application thereof |
| CN102702372B (en) * | 2010-10-28 | 2013-11-27 | 内蒙古蒙牛乳业（集团）股份有限公司 | Streptococcus thermophilus extracellular polysaccharide and preparation and detection method thereof |
| CN102465108B (en) * | 2010-10-28 | 2013-04-17 | 内蒙古蒙牛乳业（集团）股份有限公司 | Streptococcus thermophilus and application thereof |
| CN103393097A (en) * | 2013-08-02 | 2013-11-20 | 永安康健药业（武汉）有限公司 | Health food with effects of sobering and protecting liver |
| CN103937716B (en) * | 2014-04-17 | 2015-12-09 | 扬州大学 | People source lactobacillus fermentum grx07 and application thereof |
| CN105166150A (en) * | 2015-09-24 | 2015-12-23 | 佛山市三水健力宝贸易有限公司 | Black tea fungus beverage |
| KR101853603B1 (en) * | 2017-05-18 | 2018-05-02 | 주식회사 쎌바이오텍 | Composition containing of probiotics for using alcohol or acetaldehyde dehydrogenase activity |
| CN110025637A (en) * | 2018-12-29 | 2019-07-19 | 上海清微生物科技有限公司 | For the composite bacteria agent and preparation method thereof of liver detoxification |
| CN109517765A (en) * | 2019-01-11 | 2019-03-26 | 谭瑛 | A kind of streptococcus fecalis and its application |
| CN109652334A (en) * | 2019-01-11 | 2019-04-19 | 谭瑛 | A kind of complex microbial inoculum and its preparation method and application |
| JP6739602B1 (en) * | 2019-07-24 | 2020-08-12 | 雪印メグミルク株式会社 | Compositions for maintaining and/or improving memory/learning ability and foods, pharmaceuticals, feeds containing the compositions |
| CN110607253B (en) * | 2019-08-26 | 2021-12-21 | 华南理工大学 | Streptococcus thermophilus and proliferation culture method and application thereof |
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