CN115466689A - Probiotic composition for preventing and/or treating metabolic diseases and application thereof - Google Patents
Probiotic composition for preventing and/or treating metabolic diseases and application thereof Download PDFInfo
- Publication number
- CN115466689A CN115466689A CN202210122665.4A CN202210122665A CN115466689A CN 115466689 A CN115466689 A CN 115466689A CN 202210122665 A CN202210122665 A CN 202210122665A CN 115466689 A CN115466689 A CN 115466689A
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- bifidobacterium
- probiotic
- cctcc
- probiotic composition
- mice
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Abstract
The invention discloses a probiotic composition for preventing and/or treating metabolic diseases, which comprises functional probiotics, wherein the functional probiotics consist of specific bifidobacteria, can play a role in preventing and/or treating metabolic diseases, has an obvious effect of improving obesity, can improve glucose tolerance and insulin sensitivity, reduce total cholesterol in serum, reduce physiological indicators of obesity such as triglyceride in serum and the like. The invention also discloses application of the probiotic composition in preparing a probiotic product for preventing and/or treating metabolic diseases.
Description
Technical Field
The invention belongs to the technical field of probiotics, and particularly relates to a probiotic composition for preventing and/or treating metabolic diseases and application of the probiotic composition in preparation of a probiotic product for preventing and/or treating metabolic diseases.
Background
Currently, the prevalence of obesity is continuously increasing worldwide, and the World Health Organization (WHO) defines obesity as abnormal or excessive fat accumulation which may cause health damage, and is closely related to type II diabetes, hypertension, hyperlipidemia, cardiovascular diseases, liver cirrhosis and other diseases. The formation of obesity is influenced by various factors, including genetic factors, hormone use, diet, exercise and other environmental factors, and in recent years, more and more researches prove that the occurrence of obesity is closely inseparable with the intestinal flora of a host. It has been found that the intestinal flora of obese patients differs greatly from that of healthy people, and that intestinal flora (enterobacter) and bacteroides (bacteroides) are found to be more abundant than normal in the intestines of obese patients, while the abundance of bifidobacterium (bifidobacterium), oscillatoria (oscillospira), erwinia (Erwinia), vibrio succinivibrio (succinivibrio) and Alistipes (Alistipes) is significantly reduced. The improvement of obesity and its associated metabolic disorders through dietary intervention or other means to alter the composition and function of the patient's intestinal flora is an important means to effectively treat obesity that has been proposed in recent years.
Pendulum introduced Pendulum Glucose Control (PGC), the first hypoglycemic probiotic product, 6 months 2020. PGCs comprised Akkermansia muciniphila WB-STR-0001 (AKK bacteria for short), eubacterium villii WB-STR-0008 (Eubacterium hophallii), clostridium butyricum WB-STR-0006 (Clostridium butyricum), clostridium beijerinckii WB-STR-0005 (Clostridium beijerinckii), bifidobacterium infantis 100 (Bifidobacterium infantis) 5 strains and inulin 1 prebiotic in a randomized, double blind placebo control study. Compared to placebo, type II diabetics had a 0.6% decrease in A1C and a 32.5% decrease in peak blood glucose (AUC) after 12 weeks of PGC administration.
More and more researches prove that the intestinal probiotics can obviously improve metabolic diseases such as obesity, diabetes and the like in a plurality of ways such as changing the composition of intestinal bacteria, strengthening intestinal barrier, regulating intestinal immunity and the like. For example, the PGC probiotic combination can well supplement deleted strains in a patient body for a human body, and restore the micro-ecological environment of human intestinal health. However, the intestinal flora of people is greatly different from various factors such as regions, race, environment and the like, and the foreign probiotic products are not necessarily suitable for Chinese residents.
The invention aims to develop functional probiotics which are suitable for Chinese people and can play a role in preventing and/or treating metabolic diseases.
Disclosure of Invention
In view of the above, the present invention provides a probiotic composition for preventing and/or treating metabolic diseases, the probiotic composition comprises functional probiotics, and one or more than two kinds of bifidobacteria are provided. The probiotics with the specified functions are taken as active ingredients, and the probiotics has obvious effects on preventing and/or treating metabolic diseases, has obvious effect on improving obesity, can improve glucose tolerance and insulin sensitivity, reduce total cholesterol in serum, reduce triglyceride in serum and other obesity physiological indexes.
In order to achieve the purpose, the invention adopts the following technical scheme:
the invention provides a probiotic composition for preventing and/or treating metabolic diseases, which comprises any one or the combination of more than two of the strains from (1) to (5):
(1) The first Bifidobacterium faecalis (Bifidobacterium faecalis) with the preservation number of CCTCC M2022037;
(2) Second Bifidobacterium faecalis (CCTCC M2022038);
(3) Bifidobacterium longum (Bifidobacterium longum) with preservation number of CCTCC M2022039;
(4) Bifidobacterium pseudocatenulatum (Bifidobacterium pseudoocephalin) with a preservation number of CCTCC M2022040;
(5) Bifidobacterium bifidum (Bifidobacterium bifidum) with the preservation number of CCTCC M2022041;
or at least one of a bacterial strain having at least 98% genomic sequence identity to any one of the strains described in (1) - (5), or having at least 95% 16S rRNA identity to any one of the strains described in (1) - (5).
In a further scheme, in the probiotic composition, the ratio of the bacteria of the first bifidobacterium faecalis, the second bifidobacterium faecalis, the bifidobacterium longum, the bifidobacterium pseudocatenulatum and the bifidobacterium bifidum is (1-10): (1-10): (1-10): (1-10): (1-10).
In a further aspect, the probiotic composition further comprises a prebiotic component.
The invention further provides the use of a probiotic composition according to any one of the preceding claims in the preparation of a probiotic product for the prevention and/or treatment of metabolic disorders.
In a further aspect, the metabolic disease comprises obesity or type II diabetes.
In a further aspect, the probiotic product comprises a food, beverage, nutraceutical, or pharmaceutical.
The invention further provides a probiotic product for preventing and/or treating metabolic diseases, which comprises the probiotic composition as described in any one of the preceding items and a food, health product, beverage or pharmaceutically acceptable carrier, adjuvant or additive.
In a further aspect, the probiotic product is a capsule, tablet, granule, suspension, powder, or diluent.
In a further scheme, the concentration of thalli in the probiotic product is 10 8 -10 11 CFU。
The invention has the following beneficial effects:
the bacterial strains related to the probiotic composition are separated from intestinal tracts of healthy people, and the safety is high.
The probiotic composition can colonize in intestinal tract, thereby having long-term function.
In addition, the experimental result shows that the probiotic composition has good treatment effect on obesity, can improve glucose tolerance and insulin sensitivity, reduce the total cholesterol in serum, reduce triglyceride and other obesity physiological indexes in serum, and has excellent effect on preventing and/or treating metabolic diseases.
Description of biological preservation
Bifidobacterium faecium zhula-2 (Bifidobacterium faecaliszhula-2), the serial number of the strain is CCTCC M2022037, the preservation time is 1 month and 10 days in 2022 years, the preservation place is China center for type culture preservation, the concrete address is Wuhan city Wuchang region No. eight-way No. 299, the serial number of the biological preservation is CCTCC M2022037;
bifidobacterium faecium zhula-3 (Bifidobacterium faecaliszhula-3), the serial number of the strain is CCTCC M2022038, the preservation time is 1 month and 10 days in 2022 years, the preservation place is China center for type culture preservation, the concrete address is Wuhan city Wuchang region No. eight-way No. 299, the serial number of the biological preservation is CCTCC M2022038;
bifidobacterium longum zhula-4 (Bifidobacterium longumzhula-4), the serial number of the strain is CCTCC M2022039, the preservation time is 1 month and 10 days in 2022 years, the preservation place is China center for type culture preservation, the concrete address is Wuhan city Wuchang region No. eight-way No. 299, the serial number of the biological preservation is CCTCC M2022039;
bifidobacterium pseudocatenulatum zhula-5 (Bifidobacterium pseudochainzhula-5), the serial number of the strain is CCTCC M2022040, the preservation time is 2022 years, 1 month and 10 days, the preservation place is China center for type culture preservation, the concrete address is Wuhan city Wuchang region No. eight-way No. 299, the serial number of the biological preservation is CCTCC M2022040;
bifidobacterium bifidum zhula-6 (Bifidobacterium bifidumzhula b-6), the serial number of the strain is CCTCC M2022041, the preservation time is 1 month and 10 days in 2022 years, the preservation place is China center for type culture preservation, the concrete address is Wuhan city Wuchang region No. eight-way No. 299 in Hubei province, and the biological preservation serial number is CCTCC M2022041.
Drawings
Fig. 1 is a graph of the body weight change trend of mice in example 4, wherein:P<0.05，**：P<0.05；
fig. 2 is a graph of the change in blood glucose in the glucose tolerance test in mice of example 4, where:P<0.05，**：P<0.05；
fig. 3 is a graph of the change in blood glucose in the insulin sensitivity test in mice of example 4, where:P<0.05；
fig. 4 is a graph of the trend of the body weight of the obese mice in example 5, wherein:P<0.05，**：P<0.05，***：P<0.005；
fig. 5 is a graph of the change in blood glucose in the glucose tolerance test for obese mice in example 5, in which:P<0.05，**：P<0.05；
FIG. 6 is a graph showing the blood glucose changes in the insulin sensitivity test in the obese mice of example 5;
FIG. 7 is the triglyceride content in the serum of the obese mouse at the end of the experiment in example 5;
FIG. 8 is the total cholesterol level in serum of the obese mice at the end of the experiment in example 5.
Detailed Description
In order that the invention may be more fully understood, reference will now be made to the following examples. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.
The first aspect of the present invention provides a probiotic composition for preventing and/or treating metabolic diseases, which comprises specific functional probiotics, specifically, according to the embodiment of the present invention, the functional probiotics can be any one or a combination of two or more of the strains (1) to (5):
(1) The first Bifidobacterium faecalis (Bifidobacterium faecalis) has the laboratory number of zhula b-002 and the preservation number of CCTCC M2022037, and the 16sDNA sequence of the first Bifidobacterium faecalis shown as SEQ ID NO. 1;
(2) The number of the second Bifidobacterium faecalis (Bifidobacterium faecalis) is zhula b-003 in the laboratory, the preservation number is CCTCC M2022038, and the 16sDNA sequence of the second Bifidobacterium faecalis is shown as SEQ ID NO. 2;
(3) Bifidobacterium longum (Bifidobacterium longum) with laboratory number of zhula b-004 and preservation number of CCTCC M2022039, and 16sDNA sequence thereof is shown in SEQ ID NO. 3;
(4) Bifidobacterium pseudocatenulatum (Bifidobacterium pseudoceroin) with laboratory number of zhula b-005 and preservation number of CCTCC M2022040, and 16sDNA sequence shown in SEQ ID NO. 4;
(5) Bifidobacterium bifidum (Bifidobacterium bifidum) with laboratory number of zhula-006 and preservation number of CCTCC M2022041, and 16sDNA sequence shown in SEQ ID NO. 5.
In further embodiments of the invention, the functional probiotic may be at least one of the bacterial strains having at least 98% genomic sequence identity to any one of the strains described in (1) to (5), such as 98%, 98.5%, 99%, 99.5% or 100% genomic sequence identity, preferably greater than 99% genomic sequence identity; the functional probiotic bacteria can also be at least one of bacterial strains having at least 95% 16S rRNA identity with any one of the strains described in (1) - (5), for example, the 16S rRNA identity can be 95%, 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5% or 100%, and preferably the 16S rRNA identity is more than 97%. It is noted that the manner of alignment of genomic sequence identity or 16S rRNA identity described herein is well known in the art and will not be described in detail herein.
It is understood that the functional probiotics in the probiotic composition for preventing and/or treating metabolic diseases described herein may be any one of the above-mentioned strains, or a combination of any two or more of the above strains in any bacteria ratio.
As used herein, "probiotic" means a microorganism having specificity that, when swallowed or applied in sufficient quantities, is capable of inducing a functional and beneficial effect in a user that is specific to the health condition of the host. At present, most of probiotics are mainly applied to lactobacillus and Bifidobacterium, wherein Bifidobacterium (Bifidobacterium) is a gram-positive bacterium belonging to the Bifidobacterium family of Bifidobacterium order, namely a plurality of bifidobacteria belonging to the Bifidobacterium, the functional probiotics are separated from healthy human intestinal tracts, particularly, the feces of healthy volunteers are used as samples, and after separation and culture, the species are determined by PCR amplification technology and 16s rRNA sequencing analysis, and finally, the functional probiotics are obtained by in-vitro screening. The invention takes the functional probiotics as the active ingredients to form the probiotic composition, coordinates the action among probiotic microorganisms and obtains obvious effect in preventing and/or treating metabolic diseases.
Preferably, in some specific embodiments of the present invention, the functional probiotics in the probiotic composition are obtained by mixing the above five probiotics according to any ratio of bacteria, and further preferably, the ratio of bacteria of the first bifidobacterium faecalis, the second bifidobacterium faecalis, the bifidobacterium longum, the bifidobacterium pseudocatenulatum and the bifidobacterium bifidum is (1-10): (1-10): (1-10): (1-10): (1-10), more preferably, the ratio of the first bifidobacterium faecalis, the second bifidobacterium faecalis, the bifidobacterium longum, the bifidobacterium pseudocatenulatum and the bifidobacterium bifidum is 1:1:1:1:1.
in a further aspect, the probiotic composition further comprises a prebiotic component, where "prebiotic" means a dietary substance or ingredient (e.g., fiber, etc.) that is neither digested nor absorbed by the body and, when intact in the colon, selectively stimulates the growth and activity of a flora that is beneficial to the health of the individual. In some specific embodiments of the present invention, the prebiotic is preferably a carbohydrate that is not digested and absorbed by the body, and specific examples include, but are not limited to, fructo-oligosaccharide, galacto-oligosaccharide, xylo-oligosaccharide, isomalto-oligosaccharide, soy oligosaccharide, inulin, spirulina, arthrospira, etc., and further, polysaccharides (e.g., polysaccharose, caro nitrogenous polysaccharide, etc.), protein hydrolysates (e.g., casein hydrolysate, α -lactalbumin, lactoferrin, etc.), and vegetables, herbs, wild plants, etc. in natural plants, and more preferably bacterial powder or polyxylose. It is understood that the prebiotic composition may be selected by those skilled in the art based on the circumstances and will not be described in detail herein.
In a second aspect, the present invention provides the use of a probiotic composition according to the first aspect of the present invention in the manufacture of a probiotic product for the prevention and/or treatment of metabolic disorders.
Among them, the "metabolic diseases" described herein mainly include:
(i) Obesity and obesity-related disorders
(ii) Type II diabetes.
By "probiotic product" as used herein is meant a product having as an active ingredient a probiotic composition according to the first aspect of the invention, including in particular but not limited to, food, beverage, nutraceutical, pharmaceutical and the like forms.
In the third aspect of the present invention, based on the above, there is provided a probiotic product for preventing and/or treating metabolic diseases, which comprises the probiotic composition according to the first aspect of the present invention and a food, health product, beverage or pharmaceutically acceptable carrier, adjuvant or additive, wherein the "carrier", "adjuvant" or "additive" described herein is different according to different probiotic products, and can be selected conventionally in the art, and therefore will not be described in detail herein.
In some particular embodiments of the invention, the probiotic product may be a formulation prepared by mixing the probiotic composition with suitable excipients, which may be conventional in the formulation or pharmaceutical arts, such as dispersants, sweeteners, stabilizers, preservatives, etc., such as certain amino acids, vitamins or enzymes, etc.
The probiotic product of the invention can be oral capsules, suspensions, tablets, granules or powders; may be formulated with a coating, encapsulation or microencapsulation to resist gastric erosion; it can also be formulated in sustained release form to selectively release the active ingredient in the intestinal tract, particularly in the colon; furthermore, in some preferred embodiments of the present invention, the above-mentioned functional probiotics may also be mixed with suitable excipients for lyophilization and the like using techniques and equipment commonly used in lyophilization processes for pharmaceutical and/or food compositions, and since the preparation of probiotic products may be performed by means conventional in the art, they will not be specifically described herein.
Further, the specific dosage can be adjusted according to the actual situation, and in some preferred embodiments of the present invention, the total thallus concentration in the probiotic product is 1 × 10 8 -1×10 11 A CFU; more preferably, the cell concentration is 1X 10 9 Or 1X 10 10 CFU。
The present invention is illustrated below by specific examples, which are provided for illustrative purposes only and do not limit the scope of the present invention in any way, and in addition, unless otherwise specified, conditions or steps are not described in detail and the methods are conventional, and reagents and materials used are commercially available.
EXAMPLE 1 isolation of functional bacteria
Taking 10 feces samples of healthy volunteers, preserving with 20% volume fraction glycerophosphate buffer solution, and respectively diluting the feces samples to 10 -5 、10 -6 、10 -7 ；
The concentrations were plated on MRS broth (Solambio M8540), incubated anaerobically at 37 ℃ for 48 hours;
picking single clones to MRS broth for culture, performing PCR amplification by 16sRNA universal primers (upstream primer 27F;
sequencing the amplified product, determining the species through 16s RNA sequence comparison, and preserving the strain at-80 ℃ by using glycerol; selecting bifidobacteria from the separated strains to carry out in-vitro screening, and finally obtaining five potential functional probiotics which are respectively composed of bifidobacterium faecalis zhula-002, bifidobacterium faecalis zhula-003, bifidobacterium longum zhula-004, bifidobacterium pseudocatenulatum zhula-005 and bifidobacterium bifidum zhula-006.
EXAMPLE 2 culture of functional bacteria
The culture method of the five functional probiotics is consistent, and specifically comprises the following steps: the functional probiotics obtained in the frozen storage example 1 are taken out from a refrigerator at the temperature of 80 ℃ below zero, streaked in MRS broth culture medium, anaerobically cultured at the temperature of 37 ℃ until obvious monoclones appear, the monoclones are picked up to a 12ml culture tube, the MRS broth culture medium liquid culture is carried out, when the culture is carried out until the measured OD600 value is 1.0-2.0, the centrifugation is carried out for 10 minutes at 3000rpm, and bacterial thalli are collected.
EXAMPLE 3 preparation of Mixed functional bacteria combination preparation
The bacterial cells cultured in example 2 were obtained, and 2X 10 cells were collected 8 And (3) centrifuging the bacterial suspension of the CFU at 3000rpm for 10 minutes, removing the bacterial liquid supernatant, carrying out heavy suspension by using 200 mul of sterile phosphate buffer solution, and carrying out heavy suspension on five bacteria according to the ratio of 1:1:1:1:1, and centrifuging at 3000rpm for 10 minutes to collect mixed thalli;
dissolving 1.2g of bacterial powder and 0.24g of polyxylose in 20ml of sterile water to prepare sterile water containing prebiotics;
and (3) resuspending the mixed bacteria collected after centrifugation to 200 mu l by using sterile water containing prebiotics, and preparing the mixed functional bacteria combined preparation for animal experiments.
Example 4 validation of Mixed functional bacteria combination formulations to improve obesity in mice
(1) Dividing 18 SPF (specific pathogen free) grade C57/B6j mice (purchased with collecting drugs) with the age of 8-10 weeks into three groups, namely a control group (6 mice), an experimental group 1 (6 mice) and an insulin (1 IU/kg) positive control group (6 mice), respectively feeding high-fat food (Meidisen, MD12033, 60% fat kcal) from day 0 after adapting to the environment, simultaneously feeding sterile water to the control group in a gastric perfusion mode, and feeding a mixed functional bacteria combined preparation to the experimental group 1, wherein the gastric perfusion frequency is once every two days, wherein the amount of the mixed functional bacteria preparation for gastric perfusion is 1 multiplied by 10 9 CFU。
Body weights were monitored weekly with initial body weight as 100%, weekly relative body weight = real time body weight/initial body weight x 100%, and body weight curves were plotted using relative body weights. The results are shown in fig. 1, and the experimental results show that the weight growth rate of the mice in the experimental group 1 is obviously reduced compared with that of the control group, and the combined preparation of the mixed functional bacteria can obviously control the weight growth of the mice.
(2) Glucose tolerance assays were performed in mice at week 10. Specifically, the mice were fasted overnight (12-16 hours), the body weights of the mice were weighed, the amount of glucose required for the mice was calculated at a dose of 1.5g/kg, blood was taken from the tail of the mice at 0 minute, and the fasting blood glucose values of the mice were measured using a blood glucose test paper. After the mice are kept still for 30 minutes, the corresponding glucose dose is given to each mouse in a stomach irrigation mode according to the numbering sequence. The blood glucose level of the mice was measured again after 15 minutes, 30 minutes, 60 minutes, and 90 minutes from the time of gavage of the mice, and the change value of the blood glucose of the mice was plotted to indicate the degree of tolerance of the mice to glucose. The results are shown in fig. 2, and the experimental results show that the glucose tolerance of the mice of the experimental group 1 treated with the mixed functional bacteria combined preparation is obviously better than that of the control group.
(3) The insulin sensitivity test was performed at week 12. Specifically, the mice are fasted for 3-4 hours, the weight of the mice is weighed, the insulin amount required by the mice is calculated according to the dose of 1U/kg, blood is taken from the tail of the mice at 0 minute, and the fasting blood glucose value of the mice is detected by a blood glucose test paper. After the mice are kept still for 30 minutes, the corresponding insulin is given to each mouse in sequence by intraperitoneal injection according to the numbering sequence. The blood glucose level of the mice was measured again at 15 minutes, 30 minutes, 60 minutes, 90 minutes, 120 minutes from when the mice received insulin injections, and the change in blood glucose of the mice was plotted to indicate the sensitivity of the mice to insulin. The results are shown in fig. 3, and the experimental results show that the insulin sensitivity of the mice of the experimental group 1 treated with the mixed functional bacteria combination preparation is improved to some extent compared with the control group.
Example 5 validation of the Effect of the Mixed functional bacteria combination preparation on obese mice
18 SPF (specific pathogen free) C57/B6j mice (purchased from collectives) aged 8-10 weeks are divided into three groups, one group is a control group (6), one group is an experimental group 1 (6), and one group is a positive control group (6) of liraglutide (0.02 mg/kg), and after the mice are adapted to the environment, high-fat food (Meidison, MD12033, 60% fat kcal) is fed to the mice at the 10 th week until the weight reaches about 40 g. In week 0, sterile water is administered to the control group, and the mixed functional bacteria preparation is administered to the experimental group 1 once every two days, wherein the number of the mixed functional bacteria preparation for intragastric administration of the experimental group 1 is 1 × 10 9 And (4) CFU. Body weight was monitored weekly, calculated and statistically in accordance with example 4, and the results are shown in fig. 4.
Glucose tolerance measurements were performed in mice at week 15 and insulin sensitivity measurements were performed at week 18, in accordance with example 4, and the results are shown in fig. 5 and 6.
Measurement of triglyceride and total cholesterol was carried out at week 20, specifically, mouse blood was obtained 500-600. Mu.l by blood sampling from eyeball, left to stand for 30 minutes and centrifuged, and upper mouse serum was taken and measured using Triglyceride (TG) test kit (Nanjing Kangkuo, A110-1-1) and total cholesterol (T-CHO) test kit (Nanjing Kangkuo, A111-1-1), and the results are shown in FIG. 7 and FIG. 8.
As can be seen from the results in FIGS. 4-8, the weight gain of the mice in test group 1 of the intragastric mixed functional bacteria combined preparation was significantly alleviated; the glucose tolerance of the mice in the experimental group 1 is obviously superior to that of the control group, and the insulin sensitivity is improved to a certain degree; in addition, the content of triglyceride and total cholesterol in the blood serum of the mice in the experimental group 1 is reduced to a certain extent compared with that in the control group. The above experimental results show that the mixed functional bacteria combined preparation still has a therapeutic effect on obese mice.
It is to be understood that the above embodiments are merely preferred examples made for making the technical solution and technical effects of the present invention clearer. The applicant also selects one, two, three or four of the five strains in example 1 to prepare the mixed functional bacteria preparation, and carries out the verification experiments in examples 4-5, and the results show that the similar technical effects can be obtained, and the details are not specifically described herein due to space limitation. Therefore, the probiotic composition has obvious effects on preventing and/or treating metabolic diseases, has obvious effect on improving obesity, can improve glucose tolerance and insulin sensitivity, reduces total cholesterol in serum, reduces triglyceride in serum and other obesity physiological indexes.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent should be subject to the appended claims.
SEQUENCE LISTING
<110> university of science and technology in China
<120> probiotic composition for preventing and/or treating metabolic diseases and use thereof
<160> 5
<170> PatentIn version 3.3
<210> 1
<211> 1273
<212> DNA
<213> unknown
<400> 1
ggtcgcgtcc tatcagcttg atggcggggt aacggcccac catggcttcg acgggtagcc 60
ggcctgagag ggcgaccggc cacattggga ctgagatacg gcccagactc ctacgggagg 120
cagcagtggg gaatattgca caatgggcgc aagcctgatg cagcgacgcc gcgtgcggga 180
tgacggcctt cgggttgtaa accgcttttg actgggagca agcccttcgg ggtgagtgta 240
cctttcgaat aagcaccggc taactacgtg ccagcagccg cggtaatacg tagggtgcaa 300
gcgttatccg gaattattgg gcgtaaaggg ctcgtaggcg gttcgtcgcg tccggtgtga 360
aagtccatcg cttaacggtg gatccgcgcc gggtacgggc gggcttgagt gcggtagggg 420
agactggaat tcccggtgta acggtggaat gtgtagatat cgggaagaac accaatggcg 480
aaggcaggtc tctgggccgt cactgacgct gaggagcgaa agcgtgggga gcgaacagga 540
ttagataccc tggtagtcca cgccgtaaac ggtggatgct ggatgtgggg accattccac 600
ggtctccgtg tcggagccaa cgcgttaagc atcccgcctg gggagtacgg ccgcaaggct 660
aaaactcaaa gaaattgacg ggggcccgca caagcggcgg agcatgcgga ttaattcgat 720
gcaacgcgaa gaaccttacc tgggcttgac atgttcccga cagccgtaga gatacggttt 780
cccttcgggg cgggttcaca ggtggtgcat ggtcgtcgtc agctcgtgtc gtgagatgtt 840
gggttaagtc ccgcaacgag cgcaaccctc gccctgtgtt gccagcacgt cgtggtggga 900
actcacgggg gaccgccggg gtcaactcgg aggaaggtgg ggatgacgtc agatcatcat 960
gccccttacg tccagggctt cacgcatgct acaatggccg gtacaacggg atgcgacact 1020
gtgaggtgga gcggatccct taaaaccggt ctcagttcgg attggagtct gcaacccgac 1080
tccatgaagg cggagtcgct agtaatcgcg gatcagcaac gccgcggtga atgcgttccc 1140
gggccttgta cacaccgccc gtcaagtcat gaaagtgggt agcacccgaa gccggtggcc 1200
caaccttttg gggggaagcc gcttaagggg aaattctgga tgggaattaa gccgaacagg 1260
gaaaacccga aaa 1273
<210> 2
<211> 1496
<212> DNA
<213> unknown
<400> 2
tagggtttcg gatctggctc aggatgaacg ctggcggcgt gcttaacaca tgcaagtcga 60
acgggatccc aggagcttgc tcctgggtga gagtggcgaa cgggtgagta atgcgtgacc 120
gacctgcccc atacaccgga atagctcctg gaaacgggtg gtaatgccgg atgctccagt 180
tgaccgcatg gtcctctggg aaagcttttg cggtatggga tggggtcgcg tcctatcagc 240
ttgatggcgg ggtaacggcc caccatggct tcgacgggta gccggcctga gagggcgacc 300
ggccacattg ggactgagat acggcccaga ctcctacggg aggcagcagt ggggaatatt 360
gcacaatggg cgcaagcctg atgcagcgac gccgcgtgcg ggatgacggc cttcgggttg 420
taaaccgctt ttgactggga gcaagccctt cggggtgagt gtacctttcg aataagcacc 480
ggctaactac gtgccagcag ccgcggtaat acgtagggtg caagcgttat ccggaattat 540
tgggcgtaaa gggctcgtag gcggttcgtc gcgtccggtg tgaaagtcca tcgcttaacg 600
gtggatccgc gccgggtacg ggcgggcttg agtgcggtag gggagactgg aattcccggt 660
gtaacggtgg aatgtgtaga tatcgggaag aacaccaatg gcgaaggcag gtctctgggc 720
cgtcactgac gctgaggagc gaaagcgtgg ggagcgaaca ggattagata ccctggtagt 780
ccacgccgta aacggtggat gctggatgtg gggaccattc cacggtctcc gtgtcggagc 840
caacgcgtta agcatcccgc ctggggagta cggccgcaag gctaaaactc aaagaaattg 900
acgggggccc gcacaagcgg cggagcatgc ggattaattc gatgcaacgc gaagaacctt 960
acctgggctt gacatgttcc cgacagcccc agagatgggg cctcccttcg gggcgggttc 1020
acaggtggtg catggtcgtc gtcagctcgt gtcgtgagat gttgggttaa gtcccgcaac 1080
gagcgcaacc ctcgccctgt gttgccagca cgtcgtggtg ggaactcacg ggggaccgcc 1140
ggggtcaact cggaggaagg tggggatgac gtcagatcat catgcccctt acgtccaggg 1200
cttcacgcat gctacaatgg ccggtacaac gggatgcgac accgcgaggt ggagcggatc 1260
ccttaaaacc ggtctcagtt cggattggag tctgcaaccc gactccatga aggcggagtc 1320
gctagtaatc gcggatcagc aacgccgcgg tgaatgcgtt cccgggcctt gtacacaccg 1380
cccgtcaagt catgaaagtg ggtagcaccc gaagccggtg gcccaacctt ttggggggag 1440
ccgtctaagg tgagactcgt gattgggact aagtcgtaaa ggggaacccc gaaatc 1496
<210> 3
<211> 1497
<212> DNA
<213> unknown
<400> 3
attagggttt cgatctggct caggatgaac gctggcggcg tgcttaacac atgcaagtcg 60
aacgggatcc atcaggcttt gcttggtggt gagagtggcg aacgggtgag taatgcgtga 120
ccgacctgcc ccatacaccg gaatagctcc tggaaacggg tggtaatgcc ggatgctcca 180
gttgatcgca tggtcttctg ggaaagcttt cgcggtatgg gatggggtcg cgtcctatca 240
gcttgacggc ggggtaacgg cccaccgtgg cttcgacggg tagccggcct gagagggcga 300
ccggccacat tgggactgag atacggccca gactcctacg ggaggcagca gtggggaata 360
ttgcacaatg ggcgcaagcc tgatgcagcg acgccgcgtg agggatggag gccttcgggt 420
tgtaaacctc ttttatcggg gagcaagcga gagtgagttt acccgttgaa taagcaccgg 480
ctaactacgt gccagcagcc gcggtaatac gtagggtgca agcgttatcc ggaattattg 540
ggcgtaaagg gctcgtaggc ggttcgtcgc gtccggtgtg aaagtccatc gcttaacggt 600
ggatccgcgc cgggtacggg cgggcttgag tgcggtaggg gagactggaa ttcccggtgt 660
aacggtggaa tgtgtagata tcgggaagaa caccaatggc gaaggcaggt ctctgggccg 720
ttactgacgc tgaggagcga aagcgtgggg agcgaacagg attagatacc ctggtagtcc 780
acgccgtaaa cggtggatgc tggatgtggg gcccgttcca cgggttccgt gtcggagcta 840
acgcgttaag catcccgcct ggggagtacg gccgcaaggc taaaactcaa agaaattgac 900
gggggcccgc acaagcggcg gagcatgcgg attaattcga tgcaacgcga agaaccttac 960
ctgggcttga catgttcccg acggtcgtag agatacggct tcccttcggg gcgggttcac 1020
aggtggtgca tggtcgtcgt cagctcgtgt cgtgagatgt tgggttaagt cccgcaacga 1080
gcgcaaccct cgccccgtgt tgccagcgga ttatgccggg aactcacggg ggaccgccgg 1140
ggttaactcg gaggaaggtg gggatgacgt cagatcatca tgccccttac gtccagggct 1200
tcacgcatgc tacaatggcc ggtacaacgg gatgcgacgc ggcgacgcgg agcggatccc 1260
tgaaaaccgg tctcagttcg gatcgcagtc tgcaactcga ctgcgtgaag gcggagtcgc 1320
tagtaatcgc gaatcagcaa cgtcgcggtg aatgcgttcc cgggccttgt acacaccgcc 1380
cgtcaagtca tgaaagtggg cagcacccga agccggtggc ctaacccctt gtgggatgga 1440
gccgtctaag gtgaggctcg tgattgggac taagtcgtaa cagggaaaac cggaaaa 1497
<210> 4
<211> 1403
<212> DNA
<213> unknown
<400> 4
agtggcgaac gggtgagtaa tgcgtgaccg acctgcccca tacaccggaa tagctcctgg 60
aaacgggtgg taatgccgga tgctccgact cctcgcatgg ggtgtcggga aagatttcat 120
cggtatggga tggggtcgcg tcctatcagg tagtcggcgg ggtaacggcc caccgagcct 180
acgacgggta gccggcctga gagggcgacc ggccacattg ggactgagat acggcccaga 240
ctcctacggg aggcagcagt ggggaatatt gcacaatggg cgcaagcctg atgcagcgac 300
gccgcgtgcg ggatgacggc cttcgggttg taaaccgctt ttgatcggga gcaagccttc 360
gggtgagtgt acctttcgaa taagcaccgg ctaactacgt gccagcagcc gcggtaatac 420
gtagggtgca agcgttatcc ggaattattg ggcgtaaagg gctcgtaggc ggttcgtcgc 480
gtccggtgtg aaagtccatc gcttaacggt ggatctgcgc cgggtacggg cgggctggag 540
tgcggtaggg gagactggaa ttcccggtgt aacggtggaa tgtgtagata tcgggaagaa 600
caccaatggc gaaggcaggt ctctgggccg ttactgacgc tgaggagcga aagcgtgggg 660
agcgaacagg attagatacc ctggtagtcc acgccgtaaa cggtggatgc tggatgtggg 720
gcccgttcca cgggttccgt gtcggagcta acgcgttaag catcccgcct ggggagtacg 780
gccgcaaggc taaaactcaa agaaattgac gggggcccgc acaagcggcg gagcatgcgg 840
attaattcga tgcaacgcga agaaccttac ctgggcttga catgttcccg acagccgtag 900
agatatggcc tcccttcggg gcgggttcac aggtggtgca tggtcgtcgt cagctcgtgt 960
cgtgagatgt tgggttaagt cccgcaacga gcgcaaccct cgccctgtgt tgccagcacg 1020
tcatggtggg aactcacggg ggaccgccgg ggtcaactcg gaggaaggtg gggatgacgt 1080
cagatcatca tgccccttac gtccagggct tcacgcatgc tacaatggcc ggtacaacgg 1140
gatgcgacac ggcgacgtgg agcggatccc tgaaaaccgg tctcagttcg gattggagtc 1200
tgcaacccga ctccatgaag gcggagtcgc tagtaatcgc ggatcagcaa cgccgcggtg 1260
aatgcgttcc cgggccttgt acacaccgcc cgtcaagtca tgaaagtggg tagcacccga 1320
agccggtggc ctaacctttt tggatggagc cgtctaaggt gagactcgtg attgggacta 1380
agtcgaaggg ggaacgcaaa agg 1403
<210> 5
<211> 1406
<212> DNA
<213> unknown
<400> 5
agagtggcga acgggtgagt aatgcgtgac cgacctgccc catgctccgg aatagctcct 60
ggaaacgggt ggtaatgccg gatgttccac atgatcgcat gtgattgtgg gaaagatttc 120
atcggcgtgg gatggggtcg cgtcctatca gcttgttggt gaggtaacgg ctcaccaagg 180
cttcgacggg tagccggcct gagagggcga ccggccacat tgggactgag atacggccca 240
gactcctacg ggaggcagca gtggggaata ttgcacaatg ggcgcaagcc tgatgcagcg 300
acgccgcgtg agggatggag gccttcgggt tgtaaacctc ttttgtttgg gagcaagcct 360
tcgggtgagt gtacctttcg aataagcgcc ggctaactac gtgccagcag ccgcggtaat 420
acgtagggcg caagcgttat ccggatttat tgggcgtaaa gggctcgtag gcggctcgtc 480
gcgtccggtg tgaaagtcca tcgcttaacg gtggatctgc gccgggtacg ggcgggctgg 540
agtgcggtag gggagactgg aattcccggt gtaacggtgg aatgtgtaga tatcgggaag 600
aacaccgatg gcgaaggcag gtctctgggc cgtcactgac gctgaggagc gaaagcgtgg 660
ggagcgaaca ggattagata ccctggtagt ccacgccgta aacggtggac gctggatgtg 720
gggcacgttc cacgtgttcc gtgtcggagc taacgcgtta agcgtcccgc ctggggagta 780
cggccgcaag gctaaaactc aaagaaattg acgggggccc gcacaagcgg cggagcatgc 840
ggattaattc gatgcaacgc gaagaacctt acctgggctt gacatgttcc cgacgacgcc 900
agagatggcg tttcccttcg gggcgggttc acaggtggtg catggtcgtc gtcagctcgt 960
gtcgtgagat gttgggttaa gtcccgcaac gagcgcaacc ctcgccccgt gttgccagca 1020
cgttatggtg ggaactcacg ggggaccgcc ggggttaact cggaggaagg tggggatgac 1080
gtcagatcat catgcccctt acgtccaggg cttcacgcat gctacaatgg ccggtacagc 1140
gggatgcgac atggcgacat ggagcggatc cctgaaaacc ggtctcagtt cggatcggag 1200
cctgcaaccc ggctccgtga aggcggagtc gctagtaatc gcggatcagc aacgccgcgg 1260
tgaatgcgtt cccgggcctt gtacacaccg cccgtcaagt catgaaagtg ggcagcaccc 1320
gaagccggtg gcctaacccc ttgtgggatg gagccgtcta aggtgaggct cgtgattggg 1380
actaagtcga aggggggtcg gcaaac 1406
Claims (9)
1. A probiotic composition for preventing and/or treating metabolic diseases, which comprises any one or a combination of two or more of the following strains (1) to (5):
(1) Bifidobacterium bifidum (Bifidobacterium faecalis) with the preservation number of CCTCC M2022037;
(2) Bifidobacterium bifidum (Bifidobacterium faecalis) with the preservation number of CCTCC M2022038;
(3) Bifidobacterium longum (Bifidobacterium longum) with preservation number of CCTCC M2022039;
(4) Bifidobacterium pseudocatenulatum (Bifidobacterium pseudocalin) with a preservation number of CCTCC M2022040;
(5) Bifidobacterium bifidum (Bifidobacterium bifidum) with preservation number of CCTCC M2022041;
or at least one of a bacterial strain having at least 98% genomic sequence identity to any one of the strains described in (1) - (5), or having at least 95% 16S rRNA identity to any one of the strains described in (1) - (5).
2. The probiotic composition according to claim 1, characterized in that the ratio of the bacteria of said first bifidobacterium faecalis, second bifidobacterium faecalis, bifidobacterium longum, bifidobacterium pseudocatenulatum and bifidobacterium bifidum is (1-10): (1-10): (1-10): (1-10): (1-10).
3. The probiotic composition according to claim 1, characterized in that it further comprises a prebiotic ingredient.
4. Use of a probiotic composition according to any one of claims 1 to 3, for the preparation of a probiotic product for the prevention and/or treatment of metabolic disorders.
5. The use of claim 4, wherein the metabolic disorder comprises obesity or type II diabetes.
6. The use of claim 4, wherein the probiotic product comprises a food, beverage, nutraceutical, or pharmaceutical.
7. A probiotic product for the prevention and/or treatment of metabolic disorders, characterized in that it comprises a probiotic composition according to any one of claims 1 to 3 and a food, nutraceutical, beverage or pharmaceutically acceptable carrier, adjuvant or additive.
8. The probiotic product according to claim 7, characterized in that it is a capsule, tablet, granule, suspension, powder or diluent.
9. The probiotic product according to claim 7 or 8, characterized in that the concentration of bacteria in the probiotic product is 10 8 -10 11 CFU。
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Cited By (1)
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| CN111971056A (en) * | 2018-02-13 | 2020-11-20 | 普罗维瓦制药公司 | Probiotic formulations for the treatment and alleviation of metabolic and oxidative stress, inflammation and neurodegeneration |
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| CN115466689B (en) | 2023-06-16 |
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