CN112538448B - Lactobacillus with probiotic property and directionally screened from saussurea involucrata bacteria, screening method and application - Google Patents
Lactobacillus with probiotic property and directionally screened from saussurea involucrata bacteria, screening method and application Download PDFInfo
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- 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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- A61K35/74—Bacteria
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- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
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Abstract
The invention relates to lactobacillus with probiotic property, which is directionally screened from saussurea involucrata, is named as RG and classified as: lactobacillus rhamnosus, the preservation number is: CGMCCNo.20983, preservation date: 11/2/2020, depository: china general microbiological culture Collection center, Hospital No. 3 of West Lu No. 1, on the North of the Chao Yang district, Beijing, China. The invention provides the lactic acid bacteria with strong lactic acid producing, gastric acid resisting and bile resisting capabilities, the screened lactic acid bacteria can resist the severe environment in the gastrointestinal fluid digestive tract, the self-coagulation capability is strong, the oxidation resistance is strong, the bacteriostatic performance is strong, and the hemolysis activity is not generated, so that the screened lactic acid bacteria are safe and can be used for research and development of products.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to lactobacillus with probiotic characteristics, which is directionally screened from saussurea involucrate, a screening method and application.
Background
Probiotics are a class of active microorganisms that produce beneficial effects on the health of the host when given in certain amounts. Probiotics can provide valuable probiotic effects to human hosts by improving gut microbial balance, which can be significantly improved by increasing nutrient absorption. The probiotic properties of probiotics include: planting in intestinal tract, resisting oxidation, inhibiting bacteria, and improving safety. The most studied probiotics are mainly bifidobacteria and lactobacilli which are able to survive in the gastrointestinal tract. Lactic Acid Bacteria (LAB) are a generic term for a group of gram-positive, non-spore forming bacteria that can utilize carbohydrates and produce lactic acid. Lactic acid bacteria are the major bacteria in the gastrointestinal and urinary systems of animals and humans, and play an important role in maintaining and restoring health. Lactic acid bacteria are important fermentation flora and are widely used as leaven in the production of household and industrial fermented foods, such as yoghourt, pickle, fermented sausage and the like. However, the current research shows that the lactic acid bacteria have poor heat resistance and storage property due to no spore production, poor resistance to adverse environment, and particularly weak gastric acid resistance and bile resistance, so that the number of the lactic acid bacteria capable of living into the intestinal tract is small, which greatly limits the lactic acid bacteria to play beneficial functions in human bodies.
The saussurea involucrate, also known as Tibetan mushroom, is a milky white and colloidal block, is a traditional fermented food and has long-term eating history in Tibetan and Qinghai regions. Lactic acid bacteria with potential probiotic characteristics are screened from the saussurea involucrate, and are applied to fermented products, so that the fermented probiotic products which are sweet and sour and delicious and rich in various nutritional ingredients such as amino acid, vitamins and the like can be produced.
Through searching, no patent publication related to the present application has been found.
Disclosure of Invention
The invention aims to overcome the defects of weak gastric juice tolerance, poor bile resistance, less number of live bacteria entering an organism and the like of lactic acid bacteria in the prior art, and provides lactic acid bacteria with probiotic characteristics, a screening method and application thereof, which are directionally screened from saussurea involucrate.
The technical scheme adopted by the invention for solving the technical problem is as follows:
lactic acid bacteria with probiotic property are directionally screened from saussurea involucrata, are named as RG, and are classified as: lactobacillus rhamnosus, the preservation number is: CGMCC No.20983, preservation date: 11/2/2020, depository: china general microbiological culture Collection center, West Lu No. 1 Hospital No. 3, Beijing, Chaoyang, the area of the republic of China.
Moreover, the lactic acid bacteria are milky round, and the fresh surface is moist and smooth.
The method for screening the lactic acid bacteria comprises the following steps:
measuring the acid resistance of lactic acid bacteria separated from saussurea involucrate, and screening strains capable of resisting acid environments with pH2.5 and 3.0;
the survival rate of the lactobacillus screened in the step of testing and researching on the simulated gastrointestinal fluid is improved, and the number of screened live bacteria is higher than 10 6 CFU/mL lactic acid bacteria;
thirdly, the indexes of self-agglomeration, hydrophobicity, antioxidant activity, bacteriostatic performance and hemolytic activity of the lactobacillus screened in the second step are determined and researched, and the lactobacillus with probiotic characteristics is further screened out.
Use of a lactic acid bacterium as described above in food and/or pharmaceutical applications.
The invention has the advantages and positive effects that:
1. the invention provides the lactic acid bacteria with strong lactic acid producing, gastric acid resisting and bile resisting capabilities, the screened lactic acid bacteria can resist the severe environment in the gastrointestinal fluid digestive tract, the self-coagulation capability is strong, the oxidation resistance is strong, the bacteriostatic performance is strong, and the hemolysis activity is not generated, so that the screened lactic acid bacteria are safe and can be used for research and development of products.
2. The invention directionally screens out the lactic acid bacteria with probiotic characteristics from the saussurea involucrate, detects and evaluates the strains to be detected by various methods, has strong purpose and provides theoretical guidance for screening of the probiotics and further product research and development.
Drawings
FIG. 1 is a schematic diagram of lactic acid bacterium RG of the present invention;
FIG. 2 is a graph showing the results of a hemolytic test of 7 lactic acid bacteria according to the present invention;
FIG. 3 is a graph showing the change of pH and viable count of RG under different fermentation conditions; wherein A represents the change of viable count under different inoculation amounts; b represents the change of the number of viable bacteria under different sugar degrees; c represents the viable count change at different culture temperatures; d represents the change of the viable count under different inoculation amounts; e represents the change of the viable count under different sugar degrees; f represents the change in viable cell count at different culture temperatures.
Detailed Description
The following detailed description of the embodiments of the present invention is provided for the purpose of illustration and not limitation, and should not be construed as limiting the scope of the invention.
The raw materials used in the invention are conventional commercial products unless otherwise specified; the methods used in the present invention are conventional in the art unless otherwise specified.
Lactic acid bacteria with probiotic property are directionally screened from saussurea involucrata, are named as RG, and are classified as: lactobacillus rhamnosus, the preservation number is: CGMCC No.20983, preservation date: 11/2/2020, depository: china general microbiological culture Collection center, West Lu No. 1 Hospital No. 3, Beijing, Chaoyang, the area of the republic of China.
Preferably, the lactic acid bacteria are milky round, and the fresh surface is moist and smooth. As shown in fig. 1.
The method for screening the lactic acid bacteria comprises the following steps:
measuring the acid resistance of lactic acid bacteria separated from saussurea involucrate, and screening strains capable of resisting acid environments with pH2.5 and 3.0;
the survival rate of the lactobacillus screened in the step of testing and researching on the simulated gastrointestinal fluid is improved, and the number of screened live bacteria is higher than 10 6 CFU/mL lactic acid bacteria;
and thirdly, determining and researching the indexes of self-aggregation, hydrophobicity, antioxidant activity, bacteriostatic performance and hemolytic activity of the lactobacillus screened in the second step, and further screening the lactobacillus with probiotic property.
Use of a lactic acid bacterium as described above in food and/or pharmaceutical applications.
Specifically, the preparation and detection examples are as follows:
example 1 acid resistance test
Preparation of a lactic acid bacteria suspension: centrifuging the bacterium solution of the third generation activated by lactobacillus, discarding the supernatant, washing with PBS (pH7.4) for three times,adding PBS, shaking to obtain suspension, and adjusting to 1.0 × 10 concentration 8 CFU/mL(OD 595 ＝1.0)。
Inoculating lactobacillus suspension into MRS liquid culture medium with pH of 2.5 and 3.0 at 1% (v/v), culturing at 37 deg.C for 24 hr, spreading 100uL on MRS agar plate, culturing at 37 deg.C for 24 hr, and observing whether the plate has colony. The results of the experiment are shown in table 1.
Acid resistance is an important criterion for the selection of probiotic strains. Thus, ph2.5 and 3.0 were chosen as indicators for the selection of candidate probiotic strains in this experiment. As is clear from Table 1, only 10 of 19 lactic acid bacteria isolated from saussurea involucrata could grow under acidic conditions of pH2.5 and 3.0.
TABLE 1 lactic acid bacteria resistant pH2.5 and 3.0 test results
Note: "+" indicates colonies growing and "-" indicates no colonies growing.
EXAMPLE 2 gastrointestinal fluid tolerability test
Simulated artificial gastric fluid: 8g of sodium chloride, 1.15g of disodium hydrogen phosphate, 0.2g of potassium dihydrogen phosphate, 3.5g of pepsin and 1000mL of distilled water, adjusting the pH value to 3.0 by using 1mol/L hydrochloric acid, and sterilizing at 115 ℃ for 20 min.
Simulating artificial intestinal juice: 8g of sodium chloride, 1.15g of disodium hydrogen phosphate, 0.2g of potassium dihydrogen phosphate, 1g of trypsin, 3g of bile salt and 1000mL of distilled water, adjusting the pH value to 8.0 by using 1mol/L of sodium hydroxide, and sterilizing at 115 ℃ for 20 min.
(1) Selecting 10 strains of lactobacillus strain suspension resistant to pH2.5, uniformly mixing 1mL of the strain suspension with 9mL of artificial gastric juice with pH3.0, standing and culturing at 37 ℃, sampling after 0 hour and 3 hours respectively, diluting in a gradient manner, coating on an MRS agar plate, and calculating the survival rate. The results are shown in Table 2. Selecting viable count of 10 6 carrying out subsequent tolerance test on lactobacillus with cfu/ml or moreAnd (6) testing.
As shown in Table 2, after 10 strains of lactic acid bacteria were treated in simulated gastric fluid at pH3.0 for 3 hours, all the strains had a certain tolerance, but the difference was large. The survival rates of BG-1, GG-2, B5-27 and RG in artificial gastric juice are higher and are all more than 100 percent. BG-2, B5-29, NC-1 and CQ-2 have medium survival rate, the lowest survival rate of MQ-1 is 1.67%, but the number of viable bacteria is higher than 10 6 CFU/mL。
Table 2 shows the results of measuring the number of viable lactic acid bacteria in simulated gastric juice (× 10) 7 CFU/mL)
Note: the data in the table are mean ± standard deviation, and the mean is the mean of three groups of data.
(2) Uniformly mixing 1mL of lactobacillus suspension with 9mL of simulated artificial gastric juice with the pH value of 3.0, performing static culture at 37 ℃ for 3h, uniformly mixing 1mL of artificial intestinal juice with the pH value of 9mL of artificial intestinal juice with the pH value of 8.0, performing static culture at 37 ℃, sampling, diluting in a gradient manner after 0h and 4h respectively, coating the diluted sample on an MRS agar plate, and calculating the survival rate. The results are shown in Table 3. Selecting viable count of 10 6 cfu/ml of lactic acid bacteria were subjected to subsequent tolerance tests.
Survival rate (%) ═ Ns/No × 100
Wherein, No and Ns are the colony numbers of 0h and 3h or 4h simulated gastric fluid and intestinal fluid respectively.
The results of measuring the number of viable bacteria in simulated intestinal juice from 10 lactic acid bacteria selected for acid resistance are shown in Table 3. As can be seen from Table 3, after 10 strains of lactic acid bacteria were treated in simulated intestinal fluid at pH8.0 for 4 hours, the sterilized strains B5-27, B5-29 and MQ-1 all had a certain tolerance. The survival rate of BG-1 and BG-2 in the artificial intestinal juice is the highest and is 110.2 percent. The survival rates of the other strains are GG-1, NC-1, CQ-2, GG-2 and RG from large to small, and the number of the viable bacteria is higher than 10 6 CFU/mL。
Table 3 shows the results of the determination of viable count of lactic acid bacteria in simulated intestinal juice (× 10) 6 CFU/mL)
Note: the data in the table are mean ± standard deviation, and the mean is the mean of three groups of data.
Example 3 self-coagulation test
7 strains of lactic acid bacteria resistant to gastrointestinal fluids were cultured in MRS medium at 37 ℃ for 24h, centrifuged at 4000g at 4 ℃ for 15min, and washed twice with PBS after each centrifugation. The optical density 595nm of the lactic acid bacteria culture in PBS was adjusted to between 0.9 and 1 using the same buffer. Incubated at 37 ℃ for 0h, and the OD of the suspension was measured using a spectrophotometer (Kyoto Shimadzu, Japan) 595 And (5) nm. After 3h incubation the upper phase of the solution was carefully taken and the OD was measured again at 595 nm. The results are given in Table 4 below, and the self-aggregation is calculated as follows:
percent autopolymerization-1-ODt/ODo X100
Wherein ODt represents OD after 3 hours of culture 595 A value; ODo denotes OD at 0h 595 The value is obtained.
TABLE 4 self-aggregation Rate of lactic acid bacteria
Note: the data in the table are mean ± standard deviation, and the mean is the mean of three groups of data.
It is seen from Table 4 that GG-2 and RG have high self-aggregation properties of 69.13% and 65.7%, respectively. BG-1, GG-1 and NC-1 self-agglomeration capacities were at moderate levels of 37.47%, 35.59% and 32.33%, respectively. The self-coagulation ability of the rest 2 lactic acid bacteria BG-2 and CQ-2 is low, and is 28.61% and 23.99% respectively.
Example 4 hydrophobicity test
The 7 strains of lactic acid bacteria that were resistant to gastrointestinal fluids were subjected to a hydrophobicity test. The lactic acid bacteria strains were cultured in MRS at 37 ℃ for 24h, and washed twice with Phosphate Buffered Saline (PBS) at pH 7.4. The washed cells were resuspended in the same buffer and the absorbance of the cell suspension was adjusted to 0.8-1.0 ("Ao") at 595 nm. To 1mL of a solvent (n-hexane, ethyl acetate, diethyl ether and methylene chloride) was added 3mL or more of a lactic acid bacterium suspension, and the mixture was cultured at 37 ℃ for 10min and then temperature-equilibrated. The mixture was vortexed again and incubated at 37 ℃ for 3h, with phase separation without stirring. The aqueous phase (1ml) was gently removed and the absorbance "As" was measured at 595nm, the results of which are shown in Table 5. The hydrophobicity is expressed as follows:
and% hydrophobicity is calculated As [1- (As/Ao) × 100 ].
Wherein Ao represents OD before mixing 595 Value, As represents OD after mixing 595 The value is obtained. The bacterial hydrophobicity was low (0-29%), medium (30-59%) or high (60-100%), respectively.
As can be seen from Table 5, BG-2, GG-2 and RG have high hydrophobicity to ethyl acetate, 73.95%, 73.01% and 64.66%, respectively. The hydrophobicity of BG-1 and RG to diethyl ether (47.90% and 36.59%) and dichloromethane (32.36% and 32.61%) was at moderate levels. The hydrophobicity of 7 lactic acid bacteria to diethyl ether was at a moderate level, and the hydrophobicity to n-hexane was at a low level.
TABLE 5 hydrophobicity of lactic acid bacteria strains
| Hydrophobicity% | N-hexane | Ethyl acetate | Ether (A) | Methylene dichloride |
| BG-1 | 23.56±0.06 | 47.59±0.06 | 47.90±0.02 | 32.36±0.06 |
| BG-2 | 16.46±0.03 | 73.95±0.01 | 43.92±0.02 | 29.62±0.04 |
| GG-1 | 15.03±0.04 | 44.18±0.05 | 39.54±0.04 | 23.06±0.07 |
| GG-2 | 7.89±0.01 | 73.01±0.07 | 40.99±0.08 | 14.67±0.08 |
| RG | 15.35±0.05 | 64.66±0.04 | 36.59±0.04 | 32.61±0.05 |
| NC-1 | 7.24±0.03 | 16.23±0.04 | 44.61±0.04 | 24.65±0.06 |
| CQ-2 | 20.14±0.07 | 36.67±0.01 | 45.64±0.01 | 19.25±0.10 |
Note: the data in the table are mean ± standard deviation, and the mean is the mean of three groups of data.
EXAMPLE 5 Oxidation resistance test
(1) Superoxide anion scavenging activity: 4.5mL of 0.05mol/L Tris-HCl buffer (pH 8.2 containing 2mmol/L EDTA) was put in a water bath at 25 ℃ for 20min, and 2.3mL of a lactic acid bacterium suspension and 2.2mL of a 25mmol/L pyrogallol solution were added. After mixing uniformly, water bath is carried out at 25 ℃ for 4min, 1mL of 10mol/LHCL is added for stopping reaction, centrifugation is carried out at 6000r/min for 10min, supernatant fluid is taken, the light absorption value As is measured at the wavelength of 320nm, ultrapure water is used As blank control, and the result is shown in Table 6.
The clearance calculation is performed according to a formula,
clearance (%). Ao-As/Ao X100
Wherein, Ao is blank group absorbance, As is sample group absorbance.
(2) Reduction capability: the reduction capacity was determined by potassium ferricyanide method. 0.5mL of the lactic acid bacteria suspension was added to 0.2mol/L of PBS (pH 6.6) and 0.5mL of each of 1% (w/v) potassium ferricyanide solution. Reacting in water bath at 50 ℃ for 20min, and quenching. Adding 10% trichloroacetic acid 0.5mL, centrifuging at 4000r/min for 5min, collecting supernatant 1mL, adding ultrapure water and 0.1% (w/v) ferric trichloride each 1mL, mixing, standing for 10min, and adjusting to OD 700 The absorbance at nm was measured and the results are shown in Table 6. The bacterial suspension was replaced with PBS buffer as a blank control.
Reduction capacity (%) - (As-Ao)/Ao 100%
Wherein Ao is the absorbance of the blank group, and As is the absorbance of the sample group.
TABLE 6 antioxidant Activity of Lactobacillus strains
| Antioxidant activity | Superoxide anion scavenging activity% | Reduction power% |
| BG-1 | 28.53±0.01 | 67.56±0.04 |
| BG-2 | 28.46±0.00 | 73.81±0.06 |
| GG-1 | 28.11±0.01 | 61.01±0.13 |
| GG-2 | 28.13±0.01 | 86.90±0.06 |
| RG | 31.13±0.00 | 115.48±0.14 |
| NC-1 | 28.88±0.01 | 72.32±0.05 |
| CQ-2 | 23.55±0.03 | 71.13±0.05 |
Note: the data in the table are mean ± standard deviation, and the mean is the mean of three groups of data.
As is clear from Table 5, the reduction activities of 7 lactic acid bacteria were strong, and all were greater than 61%, and the reduction activity of RG was the strongest, and reached 115.48%.
EXAMPLE 6 bacteriostatic test
The antibacterial activity of 7 strains of lactic acid bacteria was determined by double-layer agar diffusion. 3 food-borne pathogens include E.coli, S.aureus and P.aeruginosa, which are used as indicator pathogens. Adding 10mL of 2% (w/v) water agar culture medium into a sterilized flat plate, placing into a sterilized Oxford cup after the water agar is solidified, adding 10% of activated third-generation LB agar culture medium into 20mL of a proper temperature 6 -10 7 And (3) fully and uniformly mixing CFU/mL indicator bacteria, pouring the mixture into a plate, taking out the Oxford cup after the upper culture medium is solidified, adding 180 mu L of 7 lactobacillus suspensions into the hole, diffusing at room temperature for 4 hours, and then culturing at 37 ℃ for 24 hours. And measuring the diameter of the transparent bacteriostatic circle by using a vernier caliper, and accurately reading to 0.01 mm. The results are given in Table 7 below.
As can be seen from Table 7, 7 strains of lactic acid bacteria all had certain bacteriostatic effects on 3 indicator bacteria, and GG-2 and RG were the best bacteriostatic effects on 3 indicator bacteria.
TABLE 7 measurement results of bacteriostatic activity of lactic acid bacteria strains
Note: the diameter of the inhibition zone comprises the aperture; the "+++" shows that the diameter of the inhibition zone is 17.0-20 mm; the "+ +" represents that the diameter of the inhibition zone is 14.0-16.9 mm; "+" indicates that the diameter of the inhibition zone is 10.0-13.9 mm; "-" indicates no bacteriostatic effect.
Example 7 hemolytic test
The hemolytic activity of 7 lactic acid bacteria was analyzed with Columbia Blood Agar (CBA) supplemented with 5% sterile defibrinated sheep blood. Streaking activated lactobacillus on CBA surface, and culturing at 37 deg.C for 24-48 h. The plates were evaluated for hemolytic activity based on different halos. No clear transparent areas were determined to be non-hemolytic. As shown in FIG. 2, the results showed that none of the 7 strains of lactic acid bacteria were hemolyzed, indicating that all of the lactic acid bacteria were safe and providing safety guarantee for further fermentation.
Application example 1: preparation of wort beverages containing the strains of the invention
(1) Preparation of wort
The mass ratio of fructus Hordei Germinatus and water is 1:4, placing in water bath kettle at 65 deg.C for 1h, cooling, filtering with gauze, boiling for 1h, cooling to room temperature, centrifuging at 4000rpm for 15min, collecting supernatant, adjusting sugar degree, packaging, and sterilizing at 115 deg.C for 20 min.
(2) Lactobacillus training test
The lactobacillus RG is gradually inoculated into different culture media with the ratio of 1:9, 2:8, 3:7, 4:6, 5:5, 6:4, 7:3, 8:2 and 9:1 of the wort and MRS culture media respectively, cultured for 24 hours at 37 ℃, and finally inoculated into pure wort for culture.
(3) Lactic acid bacteria fermentation wort test
Inoculating the trained lactobacillus RG to MRS culture medium, culturing, and regulating the bacterial concentration to 1.0 × 10 8 CFU/mL(OD 595 1.0), inoculating lactic acid bacteria RG with different bacterial concentrations into pure wort with different sugar degrees for culturing at different temperatures. As can be seen from FIG. 3, the pH of lactic acid bacteria RG reached 3.5 after 24h-36h fermentation in wort. The greater the inoculum concentration, the faster the pH drop, but 10 inoculations 8 CFU/mL began to appear weakened after 36h, 10 5 、10 6 And 10 7 CFU/mL was still in the ramp-up phase after 36 h. The sugar degree has little influence on the pH value and the viable count of the strain. The culture condition at 42 ℃ is not suitable for the growth of the lactic acid bacteria RG, and the viable count is stagnated or reduced after the culture is carried out for 36 h.
In a word, 7 strains of lactic acid bacteria are screened from 19 strains of the saussurea involucrata through gastrointestinal fluid, and are evaluated through a series of in vitro probiotic characteristic tests, the potential probiotic characteristic of the strain RG is found to be the best, and the strain RG is identified as lactobacillus rhamnosus (lactobacillus rhamnosus strain) through 16SrRNA sequencing. The lactobacillus rhamnosus RG is applied to fermented products, and is beneficial to producing fermented probiotic products which are sour, sweet, delicious and rich in various nutritional ingredients such as amino acid, vitamins and the like. The invention provides a method for screening lactobacillus strains with probiotic characteristics, which can purposefully screen lactobacillus strains with potential probiotic characteristics and probiotic application values from lactobacillus of saussurea involucrate.
The gene sequence of RG in the present invention:
r GSequence ID MT645513.1 gene sequence:
GTGTACAAGGCCCGGGAACGTATTCACCGCGGCGTGCTGATCCGCGATTACTAGCGATTCCGACTTCGTGTAGGCGAGTTGCAGCCTACAGTCCGAACTGAGAATGGCTTTAAGAGATTAGCTTGACCTCGCGGTCTCGCAACTCGTTGTACCATCCATTGTAGCACGTGTGTAGCCCAGGTCATAAGGGGCATGATGATTTGACGTCATCCCCACCTTCCTCCGGTTTGTCACCGGCAGTCTTACTAGAGTGCCCAACTAAATGCTGGCAACTAGTCATAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAACCATGCACCACCTGTCATTTTGCCCCCGAAGGGGAAACCTGATCTCTCAGGTGATCAAAAGATGTCAAGACCTGGTAAGGTTCTTCGCGTTGCTTCGAATTAAACCACATGCTCCACCGCTTGTGCGGGCCCCCGTCAATTCCTTTGAGTTTCAACCTTGCGGTCGTACTCCCCAGGCGGAATGCTTA
although the embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that: various substitutions, changes and modifications are possible without departing from the spirit and scope of the invention and the appended claims, and therefore the scope of the invention is not limited to the disclosure of the embodiments and the accompanying drawings.
Sequence listing
<110> Tianjin science and technology university
<120> lactic acid bacteria with probiotic property directionally screened from saussurea involucrate bacteria, screening method and application
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 526
<212> DNA
<213> RG Gene sequence (Unknown)
<400> 1
gtgtacaagg cccgggaacg tattcaccgc ggcgtgctga tccgcgatta ctagcgattc 60
cgacttcgtg taggcgagtt gcagcctaca gtccgaactg agaatggctt taagagatta 120
gcttgacctc gcggtctcgc aactcgttgt accatccatt gtagcacgtg tgtagcccag 180
gtcataaggg gcatgatgat ttgacgtcat ccccaccttc ctccggtttg tcaccggcag 240
tcttactaga gtgcccaact aaatgctggc aactagtcat aagggttgcg ctcgttgcgg 300
gacttaaccc aacatctcac gacacgagct gacgacaacc atgcaccacc tgtcattttg 360
cccccgaagg ggaaacctga tctctcaggt gatcaaaaga tgtcaagacc tggtaaggtt 420
cttcgcgttg cttcgaatta aaccacatgc tccaccgctt gtgcgggccc ccgtcaattc 480
ctttgagttt caaccttgcg gtcgtactcc ccaggcggaa tgctta 526
Claims (2)
1. Lactic acid bacteria with probiotic characteristics, which are directionally screened from saussurea involucrate, are characterized in that: name RG, category name: lactobacillus rhamnosus (A), (B), (C)Lactobacillus rhamnosus) The preservation number is: CGMCC No.20983, preservation date: 11/2/2020, depository: china general microbiological culture Collection center, West Lu No. 1 Hospital No. 3, Beijing, Chaoyang, the area of the republic of China.
2. Use of the lactic acid bacterium according to claim 1 in food.
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