CN114561325A - Bifidobacterium longum capable of changing bile acid content in simulated gastrointestinal tract environment and relieving constipation and application thereof - Google Patents
Bifidobacterium longum capable of changing bile acid content in simulated gastrointestinal tract environment and relieving constipation and application thereof Download PDFInfo
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- CN114561325A CN114561325A CN202210236496.7A CN202210236496A CN114561325A CN 114561325 A CN114561325 A CN 114561325A CN 202210236496 A CN202210236496 A CN 202210236496A CN 114561325 A CN114561325 A CN 114561325A
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Abstract
The invention discloses bifidobacterium longum capable of changing bile acid content in a simulated gastrointestinal environment and relieving constipation and application thereof, belonging to the technical field of microorganisms. The bifidobacterium longum CCFM1077 provided by the invention can obviously reduce the content of glycobile acid, further has the effect of relieving hyperlipidemia, and also has the effects of improving intestinal motility and relieving constipation. The capacity of the bifidobacterium longum CCFM1077 for reducing the content of the glycobile acid is obviously better than that of other three strains of bifidobacterium longum I3, J3 and B3. Therefore, the bifidobacterium longum CCFM1077 with the function of reducing the content of the glycobile acid has wide application prospect in the direction of foods and microecologics.
Description
Technical Field
The invention relates to a bifidobacterium longum capable of changing bile acid content in a simulated gastrointestinal environment and relieving constipation and application thereof, belonging to the technical field of microorganisms.
Background
Bile acid is an important component of human bile, and plays a role in digesting fat and absorbing it in the human body. More and more reports show that bile acid is an important signal molecule of human body and plays an important role in each organ. The bile acid pool of normal people is about 3-5 g, and 8-12 times of liver and intestine circulation is performed every day, wherein 95% of bile acid participates in circulation metabolism, and the rest 5% of bile acid is discharged out of the body through excrement and urine. The liver is responsible for synthesizing 5% of the bile acids daily to maintain the stability of the bile acid pool. Cholesterol is taken as a raw material in the liver, and is subjected to step metabolism by liver enzymes to finally synthesize combined primary bile acid which is discharged into the intestinal tract. Bile acids in the intestinal tract are metabolized in multiple steps by intestinal flora, constituting a rich diversity of bile acid profiles. After the conjugated bile acid enters the intestinal tract, the conjugated bile acid is metabolized into unconjugated bile acid by intestinal bacteria rich in bile salt hydrolase, such as bacteroides, clostridium, lactobacillus, bifidobacterium, listeria and the like. Further, the intestinal flora modifies the hydroxyl groups on the bile acid skeleton structure, mainly on the 7 position, and also on the 3 position, the 6 position and the 12 position, and performs the functions of dehydroxylation, dehydrogenation, isomerization and the like, and finally forms secondary bile acid. The groups involved in these metabolisms include Bacteroides, Clostridia, Escherichia, Ruminococcus, and the like. Glycobile acid, an important bile acid, causes various diseases such as liver diseases and metabolic diseases in the case of metabolic abnormality. Generally, the hepatic cells are severely damaged or bile is stagnated, and the liver fails to excrete bile acid, which causes the glycocholic acid to be higher. However, there are many reasons for the damage of liver cells, and various viral hepatitis diseases are the main reasons for the high liver function index of glycocholic acid, so that the key to the vigilance of patients is viral hepatitis. In addition, according to statistics of relevant data, liver function examination indexes of patients with acute hepatitis, chronic active hepatitis, primary liver cancer and liver cirrhosis are obviously higher than those of normal people, and the presentability is increased. Moreover, it is verified that the reason for the increase of the content of glycocholic acid in these patients is that the liver is diseased, the liver function is damaged, and the capacity of the liver cells to take glycocholic acid is reduced, so that the content of glycocholic acid in the serum is increased. In addition, if bile stagnates, the liver also fails to excrete glycocholic acid, and the reflux to the blood promotes the increase of glycocholic acid content. Therefore, the regulation of bile acid metabolism, especially the metabolism of glycobile acid, can play a role in promoting the regulation of the metabolism and the health and disease aspects of the body.
In recent years, with the progress of research methods, a great deal of research has revealed that the regulation of bile acids is associated with lipid metabolism diseases or liver diseases. For example, Degirolamo et al, published in 2014 in Cell Reports, demonstrated that supplementation with probiotic product VSL #3 can significantly alter the enterohepatic circulation of bile acids, primarily altering TCA/T β MCA and total bile acid content. Joyce et al in 2014 published in PNAS a document demonstrating that bile acid hydrolase produced by probiotics can significantly alter the composition of bile acids, primarily by altering the content of taurocholic acid.
Constipation receives increasing attention as a disease that seriously affects the quality of life. Epidemiological investigations have shown that constipation occurs in a global range of 0.7% to 79% (on average up to 16%); there is an incidence of 3% -17% in our country alone. Many causes of constipation include, for example, unbalanced diet, microbial disorders, side effects of drugs, complications due to metabolic diseases such as diabetes, genetic diseases, mechanical ileus, and neurological diseases. The treatment procedures are different for different causes of disease. For the patients with mild disease, they are often encouraged to eat more foods rich in dietary fiber such as fresh vegetables, fruits, beans, etc., but some patients with severe disease may use osmotic or irritant laxatives such as polyethylene glycol, lactulose or anthraquinone derivatives, but these laxatives often cause side effects such as dependence, even nausea, abdominal pain, diarrhea, etc., and if the patients have serious intestinal tissues with obvious pathological changes, the patients need to be treated by surgery. Therefore, the probiotics is widely accepted at present as a treatment means with small side effect, obvious relieving effect and weak dependence.
Currently, researchers at home and abroad have studied about physiological functions related to bifidobacterium, and the application of the physiological functions to the constipation relieving effect of bifidobacterium should be studied more deeply. The product can be prepared into microbial inoculum or added into food to prepare functional food, has great application prospect, and can prevent constipation and even treat constipation. The study on the bifidobacterium for relieving constipation has great influence on various aspects such as food science, microbiology, preventive medicine and the like, so that the study on the bifidobacterium for relieving constipation is necessary.
The related patents are as follows: the Mark G Kery et al patent (CN111050755A) reported that colesevelam or colesevelam hydrochloride bile acid sequestrant can relieve symptoms of patients with gastroesophageal reflux disease by lowering bile acid levels in the digestive tract. These data indicate that the intake of some drugs has a significant effect on regulating bile acid metabolism. However, regulation of glycinic acid metabolism by probiotics remains a gap.
Disclosure of Invention
In order to solve the above problems, the present invention provides a Bifidobacterium longum (Bifidobacterium longum subsp. longum) CCFM1077, which has been deposited in the Guangdong province collection of microorganisms in 2019, 9, 5 days, with the deposition number GDMCC No: 60769, the preservation address is No. 59 building 5 of No. 100 Dazhong Jie-Lu-100 Guangzhou city.
The invention also provides a microbial preparation containing the bifidobacterium longum CCFM 1077.
In one embodiment, the microbial preparation has a viable count of bifidobacterium longum CCFM1077 of not less than 1 × 106CFU/mL or 1X 106CFU/g。
The invention also provides a product for regulating bile acid metabolism, which contains the bifidobacterium longum CCFM 1077.
In one embodiment, the viable count of bifidobacterium longum CCFM1077 in the product is not less than 1 × 106CFU/mL or 1X 106CFU/g。
In one embodiment, the product is a food, pharmaceutical or nutraceutical product that is ingestible into the gastrointestinal tract.
In one embodiment, the food product comprises fermented fruits and vegetables, fermented milk, cheese, milk-containing drinks, milk powder or other food products containing the above-mentioned bifidobacterium longum.
The invention also provides application of the bifidobacterium longum CCFM1077 or the microbial preparation in preparing products for reducing the content of glycobile acid, improving the content of dehydrobile acid and/or relieving constipation.
In one embodiment, the glycobile acid includes, but is not limited to, at least one of glycofelic acid, glycohyodeoxycholic acid, glycoursodeoxycholic acid.
In one embodiment, the dehydrobile acid includes, but is not limited to, at least one of 3-dehydrocholic acid, 7-dehydrocholic acid, 12-dehydrocholic acid.
In one embodiment, the constipation relief includes, but is not limited to, increased fecal moisture content, decreased intestinal transit time.
Has the advantages that:
the bifidobacterium longum CCFM1077 provided by the invention has good tolerance in simulated gastric and intestinal fluids, can obviously change the composition of bile acid (compared with a control strain, the bifidobacterium longum CCFM1077 reduces the content of glycobile acid by nearly 8 times), and further relieves the occurrence of hyperlipidemia. And the ability of Bifidobacterium longum CCFM1077 to reduce the content of glycobile acid is significantly better than that of control strains Bifidobacterium longum strains I3, J3, B3 (disclosed in the article "Strain-Specific Effects of Bifidobacterium longum on Hypercholesterolemic rates and Potential Mechanisms", and screened from the same sample as Bifidobacterium longum CCFM 1077). Bifidobacterium longum CCFM1077 also has effect in relieving constipation induced by loperamide in mice. The bifidobacterium longum strain with the changed bile acid composition has wide application prospect in the directions of food and microecologics.
Biological material preservation
A Bifidobacterium longum (Bifidobacterium longum subsp. longum) CCFM1077, which is taxonomically named as Bifidobacterium longum subsp. longum and has been deposited in the collection center of Guangdong province microbial strains in 2019, 9 and 5 months, and the deposit number is GDMCC No: 60769, the preservation address is No. 59 building 5 of No. 100 Dazhong Jie-Lu-100 Guangzhou city.
Drawings
FIG. 1 is a colony map of Bifidobacterium longum CCFM1077 grown on mMRS medium.
FIG. 2 shows the effect of Bifidobacterium longum CCFM1077 on the content of glycobile acid in a simulated gastrointestinal environment.
FIG. 3 is a graph showing the effect of Bifidobacterium longum CCFM1077 on glycohyocholic acid content in a simulated gastrointestinal environment.
FIG. 4 is a graph showing the effect of Bifidobacterium longum CCFM1077 on glycohyodeoxycholic acid content in a simulated gastrointestinal environment.
FIG. 5 is a graph showing the effect of Bifidobacterium longum CCFM1077 on the content of glycoursodeoxycholic acid in simulated gastrointestinal environment.
FIG. 6 is a graph showing the effect of Bifidobacterium longum CCFM1077 on the content of 3-dehydrocholic acid in simulated gastrointestinal environment.
FIG. 7 is a graph of the effect of Bifidobacterium longum CCFM1077 on the content of 7-dehydrocholic acid in simulated gastrointestinal environments.
FIG. 8 is a graph of the effect of Bifidobacterium longum CCFM1077 on the content of 12-dehydrocholic acid in simulated gastrointestinal environments.
FIG. 9 shows the effect of Bifidobacterium longum CCFM1077 on loperamide-induced reduction in the frequency of defecation in mice.
FIG. 10 is a graph showing the effect of Bifidobacterium longum CCFM1077 on the reduction of loperamide-induced intestinal transit time in mice.
FIG. 11 shows the effect of Bifidobacterium longum CCFM1077 on loperamide-induced reduction in fecal water content in mice.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings.
The media involved in the following examples are as follows:
MRS liquid medium: 10g/L of peptone, 10g/L of beef extract, 5g/L of yeast extract, 20g/L of glucose, 2g/L of anhydrous sodium acetate, 2g/L of citric acid hydrogen diamine and K2HPO4·3H2O 2.6g/L，MgSO4·7H2O 0.5g/L，MnSO4·H20.25g/L of O, 1g/L of cysteine hydrochloride, 801 g/L of Tween-and 1000g/L of distilled water.
MRS solid medium: 10g/L of peptone, 10g/L of beef extract, 5g/L of yeast extract, 20g/L of glucose, 2g/L of anhydrous sodium acetate, 2g/L of citric acid hydrogen diamine and K2HPO4·3H2O 2.6g/L，MgSO4·7H2O 0.5g/L，MnSO4·H20.25g/L of O, 1g/L of cysteine hydrochloride, 801 g/L of Tween-801, 20g/L of agar and 1000g/L of distilled water.
The preparation of the bacteroides vulgatus suspension referred to in the following examples was as follows:
streaking on MRS solid culture medium, and carrying out anaerobic culture at 37 ℃ for 48h to obtain single colony; selecting a single colony, inoculating the single colony in an MRS liquid culture medium, culturing for 18h at 37 ℃ for activation, and continuously activating for two generations to obtain an activation solution; inoculating the activated liquid into an MRS liquid culture medium according to the inoculation amount of 2% (v/v), and culturing for 18h at 37 ℃ to obtain a bacterial liquid; centrifuging the bacterial liquid at 8000g for 10min to obtain Bifidobacterium longum thallus; washing Bifidobacterium longum with physiological saline, and suspending in 200g/L glycerol solution (containing 1g/L cysteine hydrochloride) to bacterial concentration of 1 × 1010CFU/mL to obtain a bacterial suspension, and storing the bacterial suspension at-80 ℃ for later use.
Example 1: separation and screening of bifidobacterium longum CCFM111077
(l) 1g of fresh faeces of a healthy person was taken. After gradient dilution, coating on mMRS solid culture medium, and culturing for 72 hours at 37 ℃ in an anaerobic environment;
(2) observing and recording the colony morphology, selecting colonies, and streaking and purifying;
(3) the colonies were gram-stained in MRS liquid medium at 37 ℃ for 48 hours, and the colony morphology was recorded.
(4) Removing gram-negative bacteria strains and gram-positive cocci from the colonies, and selecting to obtain gram-positive bacilli.
(5) After catalase analysis, catalase-positive strains were discarded, and catalase-negative strains were retained.
(II) preliminary identification of Bifidobacterium: fructose-6-phosphate phosphoketolase assay
(1) Culturing the lactic acid bacteria obtained by screening in the step (I) in a liquid mMRS culture solution for 24h, and then centrifuging the lmL culture at 8000rpm for 2 min;
(2) using 0.05M KH of pH6.5 containing 0.05% (mass percent) cysteine hydrochloride2PO4Washing the solution for two times;
(3) resuspending in 200. mu.L of the above phosphate buffer solution to which 0.25% (mass%) Triton X-100 was added;
(4) adding 50 mu L of mixed solution of sodium fluoride with the concentration of 6mg/mL and sodium iodoacetate with the concentration of 10mg/mL and 50 mu L of fructose-6-phosphate with the concentration of 80mg/mL, and incubating for 1h at 37 ℃;
(5) adding 300 μ L hydroxylamine hydrochloride with concentration of 0.139g/mL and pH of 6.5, and standing at room temperature for 10 min;
(6) separately, 200. mu.L of 15% (mass percent) trichloroacetic acid and 4M HCI were added;
(7) when 200. mu.L of 0.1M HCI containing 5 mass% of ferric trichloride was added, the system rapidly turned red, i.e., it was positive for F6PPK, and it was preliminarily judged to be Bifidobacterium.
(III) molecular biological identification of bifidobacteria:
(l) Extracting a single-bacterium genome: culturing the bifidobacterium obtained by screening in the step (II) overnight, taking the overnight-cultured bacterium suspension lmL in a 1.5mL centrifuge tube, centrifuging for 2min at 10000rpm, and removing the supernatant to obtain thalli; purging the thallus with lmL sterile water, centrifuging at 10000rpm for 2min, and removing the supernatant to obtain thallus; adding 200 μ L SDS lysate, and water bathing at 80 deg.C for 30 min; adding 200 mu of phenol-chloroform solutionPutting the L in a thallus lysate, wherein the composition and volume ratio of a phenol-chloroform solution are Tris saturated phenol, chloroform and isoamylol (25: 24: 1), reversing and mixing uniformly, centrifuging at 12000rpm for 5-10min, and taking 200 mu L of supernatant; adding 400 μ L of glacial ethanol or glacial isopropanol into 200uL of supernatant, standing at-20 deg.C for 1h, centrifuging at 12000rpm for 5-10min, and removing supernatant; adding 500 μ L70% (volume percentage) of glacial ethanol, resuspending the precipitate, centrifuging at 12000rpm for 1-3min, and discarding the supernatant; oven drying at 60 deg.C, or naturally air drying; 50 μ L ddH2Re-dissolving the precipitate with O for PCR;
(2)16S rDNA PCR：
A. bacterial 16S rDNA 50 μ LPCR reaction system:
10 × Taq buffer, 5 μ L; dNTP, 5. mu.L; 27F, 0.5 μ L; 1492R, 0.5 μ L; taq enzyme, 0.5. mu.L; template, 0.5 μ L; ddH2O，38μL。
PCR conditions:
95℃5min；95℃10s；55℃30s；72℃30s；step2-4 30×；72℃5min；12℃2min；
C. preparing 1% agarose gel, mixing the PCR product with 10000 × loading buffer, loading 2 μ L sample, running at 120V for 30min, and performing gel imaging;
D. the obtained PCR product is sent to a professional sequencing company, and the obtained sequencing result is searched and compared with the similarity in GeneBank by using BLAST to be identified as the bifidobacterium longum.
(3) Whole genome sequencing
The extracted whole genome is sent to a professional sequencing company, the whole genome of the strain is sequenced by a second-generation sequencer, the obtained sequence result is searched and compared with similarity in GeneBank by using BLAST, and the identification result shows that the average nucleotide consistency (ANI) of B.longum CCFM1077 and a standard strain Bifidobacterium longum NCC2705 genome is 98.3 percent and is higher than a strain identification seed level threshold value (more than or equal to 95 percent), and the result shows that the strain is a new Bifidobacterium longum strain. Strains I3, B3, J3, s7 were screened from stool samples of healthy adults and identified as Bifidobacterium longum, and subsequent experiments were used as control strains (strains I3, B3, J3 have been disclosed in the article "Strong-Specific Effects of Bifidobacterium on Hypercholesterolemic rates and functional Mechanisms"). The strain is preserved at the temperature of-80 ℃ for later use.
Example 2: bifidobacterium longum subspecies longum CCFM1077 has good tolerance to simulated gastrointestinal fluid
Inoculating the frozen and preserved bifidobacterium longum subspecies CCFM1077 into an mMRS culture medium (MRS culture medium + 0.05% cysteine hydrochloride), carrying out anaerobic culture at 37 ℃ for 48h, carrying out subculture for 2-3 times by using the mMRS culture medium, mixing 1mL of the culture medium of the bifidobacterium longum subspecies CCFM1077 with 9.0mL of artificial simulated gastric juice (the mMRS culture medium containing 1% pepsin and having the pH of 2.5), carrying out anaerobic culture at 37 ℃, sampling at 0h, 0.5h, 1h and 2h respectively, carrying out pouring culture by using the mMRS agar culture medium, carrying out plate colony counting, measuring the viable count and calculating the survival rate.
The survival rate is the ratio of the log of viable bacteria at sampling to the log of viable bacteria at 0h in the culture broth, and is expressed in%. Adding 1mL of culture solution of Bifidobacterium longum subspecies CCFM1077 into 9mL of artificial simulated intestinal fluid (mMRS culture medium containing 0.3% of cholate, 1% of trypsin and pH 8.0), anaerobically culturing at 37 deg.C, sampling at 0h, 0.5h, 1h and 2h respectively, pouring and culturing with mMRS agar culture medium, counting plate colony, measuring viable count and calculating survival rate. The survival rate is the ratio of the log of viable bacteria at sampling to the log of viable bacteria at 0h in the culture broth, and is expressed in%. The results of the experiment are shown in tables 1 and 2. The result shows that the bifidobacterium longum subspecies longum CCFM1077 has better tolerance to the artificial gastrointestinal fluid.
TABLE 1 tolerance of Bifidobacterium longum subspecies longum CCFM1077 in simulated gastric juice
TABLE 2 tolerance of Bifidobacterium longum subspecies longum CCFM1077 in artificial simulated intestinal fluid
Example 3: bifidobacterium longum CCFM1077 function of reducing content of glycobile acid in simulated gastrointestinal fluid environment
Individual cells from freshly streaked and grown solid media were inoculated into 10mL of MRS liquid media and incubated in an anaerobic incubator at 37 ℃ for 18 hours. Normalization to OD for each culture broth600After a value of 1, the cells were prepared and washed twice with PBS. The washed cells were resuspended, and 1ml of cells were added to LB broth medium containing 0.5% cholecystogenic bile of swine. Adding the missing human bile acid into LB culture medium added with the pig bile acid, which comprises the following steps: cholic acid (cholic acid), α -muricholic cholic acid, β -muricholic cholic acid, ω -muricholic cholic acid). 1mL of the concentrate is 1X 107CFU/mL of the bacterial suspension was incubated with bile for 1.5 hours. Simultaneously, deuterated internal standards of cholic acid and chenodeoxycholic acid are added. Followed by centrifugation in a centrifuge under the following conditions: 10000g, 10 min. The supernatant was taken and extracted with 1 volume of methanol (split into 2 tubes) to a final concentration of 50% methanol. The mixture was allowed to stand at-20 ℃ for 30 minutes and vortexed. Then dried under vacuum under nitrogen and re-extracted with acetonitrile and formic acid (1 ml). In this step, the samples are pooled together. The extract was centrifuged and dried and resuspended in 150. mu.l of 50% methanol on the day of MS detection or stored dry at-20 ℃ until the day of UPLC-MS detection. The results show that CCFM1077 can significantly reduce the content of glycobile acid (1858.4ng/mL), compared with a blank control group (15156.4ng/mL), the content of glycobile acid is reduced by 87.7% (figure 2), the content of glycobile acid is reduced from 440317.95ng/mL to 20112.2ng/mL and is reduced by 95.4% (figure 3), the content of glycodeoxycholic acid is reduced from 348530.8ng/mL to 3350.6 ng/mL and is reduced by 99.1% (figure 4), and the content of glycoursodeoxycholic acid is reduced from 5061.4ng/mL to 928.0ng/mL and is reduced by 81.7% (figure 5).
Example 4: bifidobacterium longum CCFM1077 for improving dehydrobile acid content in simulated gastrointestinal fluid environment
Single mycelia from freshly streaked and grown solid medium were inoculated into 10mL of MRS liquid medium and placed under anaerobic conditions at 37 deg.CAnd (4) an oxygen incubator for 18 hours. Normalization to OD for each culture broth600After a value of 1, cells were prepared and washed twice with PBS. The final suspension was resuspended in LB broth containing 0.5% cholecystogenic bile from swine. The corresponding missing bile acids, cholic acid (alpha, beta, omega) and their conjugated forms, were added to the master mix bile acid LB. The cells were incubated with bile for 1.5 hours. Deuterated internal standards of cholic acid and chenodeoxycholic acid are added at the same time. Then, centrifuging in a centrifuge under the following conditions: 10000g, 10 min. The supernatant was taken and extracted with 1 volume of methanol (split into 2 tubes) to a final concentration of 50% methanol. The mixture was allowed to stand at-20 ℃ for 30 minutes and vortexed. Then dried under vacuum under nitrogen and re-extracted with acetonitrile and formic acid (1 ml). In this step, the samples are pooled together. The extract was centrifuged and dried and resuspended in 150. mu.l of 50% methanol on the day of MS detection or stored at-20 ℃ until dry on the day of UPLC-MS detection. The results show that bifidobacterium longum CCFM1077 can increase the content of 3-dehydrocholic acid from 8.8ng/mL to 1331.5ng/mL by 150 times (FIG. 6), increase the content of 7-dehydrocholic acid from 16.6ng/mL to 180.7ng/mL by 9.9 times (FIG. 7) and increase the content of 12-dehydrocholic acid from 120.0ng/mL to 408.1ng/mL by 2.4 times (FIG. 8) compared with the blank control group.
Example 5: bifidobacterium longum CCFM1077 for preparing fermented milk
Bifidobacterium longum CCFM1077 can be used for preparing fermented cow milk, and the specific preparation process of cow milk is as follows:
(1) inoculating the secondary purified culture solution of bifidobacterium longum CCFM1077 obtained in example 1 into a culture medium at an inoculation amount of 3% (v/v), and culturing at 37 ℃ for 18h to obtain a bacterial solution; centrifuging the bacterial liquid to obtain bacterial sludge; washing the bacterial sludge with phosphate buffer solution of pH7.2 for 3 times, and resuspending with protectant to 1 × 1010CFU/mL to obtain a suspension; pre-culturing the suspension at 37 deg.C for 60min, and lyophilizing to obtain starter;
the preparation method of the culture medium comprises the following steps: dissolving 10% of enzyme hydrolysis skim milk, 0.5% of glucose, 1.5% of tryptone and 0.3% of yeast extract by using 87.7% of water based on the total weight of the culture medium, and then adjusting the pH of the solution to 6.8 to obtain a culture medium;
the components of the protective agent comprise: 100g/L of skimmed milk powder, 30mL/L of glycerol, 100g/L of maltodextrin, 150g/L of trehalose and 10g/L L-sodium glutamate;
(2) sterilizing skimmed milk at 95 deg.C for 20min, and cooling to 4 deg.C to obtain raw material; adding the leaven prepared in the step (1) into the raw materials until the concentration is not less than 1 x 106CFU/mL to obtain cow milk (the cow milk needs to be refrigerated at 4 ℃).
Example 6: bifidobacterium longum CCFM1077 for preparing fermented soybean milk
Bifidobacterium longum CCFM1077 can be used for preparing soybean milk, and the specific preparation process of the soybean milk is as follows:
(1) inoculating the secondary purified culture solution of bifidobacterium longum CCFM1077 obtained in example 1 into a culture medium at an inoculation amount of 3% (v/v), and culturing at 37 ℃ for 18h to obtain a bacterial solution; centrifuging the bacterial liquid to obtain bacterial sludge; washing the bacterial sludge with phosphate buffer solution of pH7.2 for 3 times, and resuspending with protectant to 1 × 1010CFU/mL to obtain a suspension; pre-culturing the suspension at 37 deg.C for 60min, and lyophilizing to obtain starter;
the preparation method of the culture medium comprises the following steps: dissolving 10% of skimmed milk, 0.5% of glucose, 1.5% of tryptone and 0.3% of yeast extract with 87.7% of water based on the total weight of the culture medium, and then adjusting the pH to 6.8 to obtain a culture medium;
the components of the protective agent comprise: 100g/L of skimmed milk powder, 30mL/L of glycerol, 100g/L of maltodextrin, 150g/L of trehalose and 10g/L L-sodium glutamate;
(2) soaking soybean at 80 deg.C for 2 hr, removing soybean hull to obtain peeled soybean; draining peeled semen glycines, soaking in water, adding boiling water, and pulping to obtain soybean milk; keeping the temperature of the soybean milk at a temperature higher than 80 ℃ for 12min to obtain cooked soybean milk; filtering the cooked soybean milk with a 150-mesh screen and then carrying out centrifugal separation to obtain coarse soybean milk; heating the coarse soybean milk to 140-150 ℃, and then quickly introducing the coarse soybean milk into a vacuum cooling chamber for vacuumizing, so that peculiar smell substances in the coarse soybean milk are quickly discharged along with water vapor to obtain cooked soybean milk; cooling the cooked soy milk to aboutAdding the leaven prepared in step (1) into cooked soybean milk at 37 deg.C to reach concentration of not less than 1 × 106CFU/mL to obtain soybean milk (the soybean milk should be stored at 4 deg.C under refrigeration).
Example 7: bifidobacterium longum CCFM1077 for preparing fruit and vegetable beverage
Bifidobacterium longum CCFM1077 can be used for preparing vegetable beverage, and the vegetable beverage is prepared by the following steps:
(1) inoculating the secondary purified culture solution of bifidobacterium longum CCFM1077 obtained in example 1 into a culture medium at an inoculation amount of 3% (v/v), and culturing at 37 ℃ for 18h to obtain a bacterial solution; centrifuging the bacterial liquid to obtain bacterial sludge; washing the bacterial sludge with phosphate buffer solution of pH7.2 for 3 times, and resuspending the bacterial sludge with protective agent to 1 × 1010CFU/mL to obtain a suspension; pre-culturing the suspension at 37 deg.C for 60min, and lyophilizing to obtain starter;
the preparation method of the culture medium comprises the following steps: dissolving 10% of enzyme hydrolysis skim milk, 0.5% of glucose, 1.5% of tryptone and 0.3% of yeast extract by using 87.7% of water based on the total weight of the culture medium, and then adjusting the pH of the solution to 6.8 to obtain a culture medium;
the components of the protective agent comprise: 100g/L of skimmed milk powder, 30mL/L of glycerol, 100g/L of maltodextrin, 150g/L of trehalose and 10g/L L-sodium glutamate;
(2) cleaning fresh vegetables and squeezing to obtain vegetable juice; thermally sterilizing the vegetable juice at 140 deg.C for 2 s to obtain sterilized vegetable juice; cooling the sterilized vegetable juice to about 37 deg.C, adding the starter prepared in step (1) into the sterilized vegetable juice to a concentration of not less than 1 × 106CFU/mL to obtain vegetable beverage (the vegetable beverage needs to be stored at 4 deg.C under refrigeration).
Example 8: bifidobacterium longum CCFM1077 for preparing capsule product
Bifidobacterium longum CCFM1077 can be used for preparing capsule products, and the specific preparation process of the capsule products is as follows:
(1) inoculating the secondary purified culture solution of bifidobacterium longum CCFM1077 obtained in example 1 into a culture medium at an inoculation amount of 3% (v/v), and culturing at 37 ℃ for 18h to obtain a bacterial solution; will be provided withCentrifuging the bacterial liquid to obtain bacterial sludge; washing the bacterial sludge with phosphate buffer solution of pH7.2 for 2 times, and suspending with skimmed milk to concentration of 2 × 1010CFU/mL to obtain a suspension;
(2) adding the suspension prepared in the step (1) into a sodium alginate solution with the concentration of 3 percent to the concentration of 2 multiplied by 109Fully stirring after CFU/mL to uniformly disperse cells of the bifidobacterium longum CCFM1077 in the sodium alginate solution to obtain a mixed solution; extruding the mixed solution into a calcium chloride solution with the concentration of 2% to form colloidal particles; standing and solidifying the formed colloidal particles for 30min, and filtering and collecting the colloidal particles; freeze-drying the collected colloidal particles for 48 hours to obtain powder; and filling the powder into a medicinal capsule to obtain a capsule product.
Example 9: bifidobacterium longum CCFM1077 for preparing fermented milk product
Bifidobacterium longum CCFM1077 can be used for preparing fermented milk, and the specific preparation process of the fermented milk is as follows:
(1) inoculating the secondary purified culture solution of bifidobacterium longum CCFM1077 obtained in example 1 into a culture medium at an inoculation amount of 3% (v/v), and culturing at 37 ℃ for 18h to obtain a bacterial solution; centrifuging the bacterial liquid to obtain bacterial sludge; washing the bacterial sludge with phosphate buffer solution of pH7.2 for 3 times, and resuspending the bacterial sludge with protective agent to 1 × 1010CFU/mL to obtain a suspension; pre-culturing the suspension at 37 deg.C for 60min, and lyophilizing to obtain lyophilized powder with thallus concentration of 1 × 109CFU/g；
The preparation method of the culture medium comprises the following steps: dissolving 10% of enzyme hydrolysis skim milk, 0.5% of glucose, 1.5% of tryptone and 0.3% of yeast extract by using 87.7% of water based on the total weight of the culture medium, and then adjusting the pH of the solution to 6.8 to obtain a culture medium;
the components of the protective agent comprise: 100g/L skimmed milk powder, 30mL/L glycerin, 100g/L maltodextrin, 150g/L trehalose, and 10g/L L-sodium glutamate;
(2) mixing lyophilized powder with commercial dry powder leaven Lactobacillus bulgaricus (thallus concentration of 1 × 10)9CFU/g) and commercial dry powder starter Streptococcus thermophilus (cell concentration 1X 10)9CFU/g) according to a mass ratio of 1:1:1Mixing to obtain a starter culture, wherein the bacterial concentration of Lactobacillus gasseri and Streptococcus thermophilus in the starter culture is 2%;
(3) adding sugar into fresh milk to reach a concentration of 5% to obtain a mixed solution; homogenizing the mixed solution at 65 deg.C and 20MPa, and sterilizing at 95 deg.C for 5min to obtain fermentation raw material; cooling the fermentation raw material to 35 ℃, inoculating the starter prepared in the step (2) into the fermentation raw material in an inoculation amount of 0.03% (v/v), and fermenting at the temperature of 35 ℃ for 16h to obtain fermented milk; after curdling the fermented milk at 42 ℃, refrigerating the fermented milk at 4 ℃ for 24h for after-ripening to obtain a fermented milk finished product.
Example 10: application of bifidobacterium longum CCFM1077
Bifidobacterium longum CCFM1077 can be used for preparing tablets, and the specific preparation process of the tablets is as follows:
(1) inoculating the secondary purified culture solution of bifidobacterium longum CCFM1077 obtained in example 1 into a culture medium at an inoculation amount of 3% (v/v), and culturing at 37 ℃ for 18h to obtain a bacterial solution; centrifuging the bacterial liquid to obtain bacterial sludge; washing the bacterial sludge with phosphate buffer solution of pH7.2 for 3 times, and resuspending the bacterial sludge with protective agent to 1 × 1010CFU/mL to obtain a suspension; pre-culturing the suspension at 37 deg.C for 60min, and lyophilizing to obtain bacterial powder;
the preparation method of the culture medium comprises the following steps: dissolving 10% of enzyme hydrolysis skim milk, 0.5% of glucose, 1.5% of tryptone and 0.3% of yeast extract by using 87.7% of water based on the total weight of the culture medium, and then adjusting the pH of the solution to 6.8 to obtain a culture medium;
the components of the protective agent comprise: 100g/L of skimmed milk powder, 30mL/L of glycerol, 100g/L of maltodextrin, 150g/L of trehalose and 10g/L L-sodium glutamate;
(2) weighing 25.7 parts by weight of the fungus powder prepared in the step (1), 55.0 parts by weight of starch, 4.5 parts by weight of cellulose derivative, 12.0 parts by weight of sodium carboxymethyl starch, 0.8 part by weight of talcum powder, 1.0 part by weight of cane sugar and 1.0 part by weight of water to obtain a raw material; mixing the raw materials to obtain wet granules; the wet granules were tableted with a tablet press of pharmaceutical machinery of south-central institute and dried with a small-sized drug dryer of yikang traditional Chinese medicine machinery ltd, qingzhou to obtain tablets.
The results of adding the products prepared in examples 5-10 into a simulated gastrointestinal fluid environment show that the products containing bifidobacterium longum CCFM1077 have the effect of regulating the content of glycobile acid and/or dehydrobile acid.
Example 11: bifidobacterium longum CCFM1077 relieves constipation-associated symptoms induced by loperamide
After 50 male BALB/c mice aged 8 weeks were acclimated for 1 week, 10 mice per group were randomly divided into 5 groups including a blank control (gavage normal saline), a constipation model group (gavage loperamide and normal saline), a positive drug (phenolphthalein) control group (gavage loperamide and 7mg/mL phenolphthalein solution), Bifidobacterium longum S7 and Bifidobacterium longum CCFM1077 groups (gavage loperamide and 1X 10M9CFU/mL of a bacterial suspension of bifidobacterium longum), and gavage for 14 days. Mice were housed in constant temperature and humidity (temperature: 25. + -. 2 ℃ C.; humidity: 55% + -5%) animal house IVC cages, with 12h dark and 12h light cycles, and standard commercial mouse food and free drinking water was given to the mice during the experiment. Animal protocol approved by the ethical committee of south of the Yangtze university (JN.No20180615b0800804[ 159)]) The experimental procedures were carried out according to the guidelines for laboratory animals (Directive 2010/63/EU) established in the European Union.
Collecting the feces of the mice before the experiment is finished, injecting a 1% sodium pentobarbital solution into the abdominal cavity of the mice for anesthesia after the experiment is finished, collecting blood and colon contents, measuring the propelling length of ink, and freezing the sample in a refrigerator at-80 ℃ for measuring other indexes. Collecting mouse feces at 8-10 points in the morning on 0, 7 and 14 days, independently placing each mouse in a clean IVC cage box, collecting the feces of the mice in time, recording the number of the feces particles, placing the feces particles in an ice box for storage, recording the dry weight of the feces after freeze drying, and calculating the moisture content of the feces by adopting the following formula: stool water content (%): (wet stool weight-dry stool weight) ÷ wet stool weight x 100%. And (3) detecting the first black stool time of the mice on the 14 th day, except for infusing 0.2mL of physiological saline to the mice of the blank control group, infusing 0.2mL of loperamide to the mice of the other groups, infusing 0.25mL of ink to the mice of the blank control group and the mice of the constipation group after 1h, infusing 0.25mL of ink containing respective infused contents to the mice of the bifidobacterium longum CCFM1077 group, the bifidobacterium longum S7 group and the positive drug control group, placing the infused mice in a clean IVC cage paved with white filter paper, and recording the time of discharging the first black stool after the ink is infused to each mouse.
From fig. 9-11, it can be seen that bifidobacterium longum CCFM1077 group significantly increased stool water content, defecation frequency and intestinal transit time compared to constipation model group, wherein the stool water content after bifidobacterium longum CCFM1077 group was increased by 67% (P <0.0001) compared to constipation model group, and the stool water content after bifidobacterium longum CCFM1077 group was gavaged was 62.8%; after the bifidobacterium longum is perfused with the stomach CCFM1077, the intestinal transit time (167min) is shortened to a certain extent, and is shortened by 36.7 percent compared with a constipation model group; the defecation frequency is obviously reduced after the bifidobacterium longum CCFM1077 is perfused into the stomach (P < 0.001). Meanwhile, compared with a control group (the water content of the excrement is 50.8 percent, and the intestinal transit time is 178min), the water content of the excrement is improved by 23.7 percent, and the intestinal transit time is reduced by 6.3 percent. On the three appearance indexes, the action effect of the bifidobacterium longum CCFM1077 is better than that of the bifidobacterium longum S7. In conclusion, bifidobacterium longum CCFM1077 showed good constipation relieving effect from the appearance index.
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (10)
1. Bifidobacterium longum (Bifidobacterium longum subsp. longum) CCFM1077, which has been deposited in the collection of microorganisms of the Guangdong province in 2019, 9 and 5 months, with the deposit number GDMCC No: 60769.
2. a microbial preparation comprising Bifidobacterium longum CCFM1077 according to claim 1.
3. A product for regulating bile acid metabolism comprising Bifidobacterium longum as claimed in claim 1 or a microbial preparation as claimed in claim 2 or 3.
4. A product according to claim 3, wherein the Bifidobacterium longum has a viable count of not less than 1 x 106CFU/mL or 1X 106CFU/g。
5. The product of claim 4, wherein the product is a food, pharmaceutical or nutraceutical product that is ingestible into the gastrointestinal tract.
6. The product of claim 6, wherein the food product is a fermented fruit or vegetable, a fermented milk, a cheese, a milk-containing drink, or a powdered milk.
7. Use of a bifidobacterium longum CCFM1077 according to claim 1 or a microbial preparation according to claim 2 in the manufacture of a product for reducing the content of glycobile acid, increasing the content of dehydrobile acid and/or alleviating constipation.
8. The use of claim 7, wherein said glycobile acid includes, but is not limited to, at least one of glycofelic acid, glycohyodeoxycholic acid, glycoursodeoxycholic acid.
9. The use of claim 7, wherein said dehydrobile acid includes, but is not limited to, at least one of 3-dehydrocholic acid, 7-dehydrocholic acid, 12-dehydrocholic acid.
10. The use according to any one of claims 7 to 9, wherein the product includes but is not limited to functional food, health care products or pharmaceuticals.
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| CN111073828A (en) * | 2019-11-19 | 2020-04-28 | 江南大学 | Bifidobacterium longum subspecies longum and application thereof |
| CN113068837A (en) * | 2021-04-06 | 2021-07-06 | 江南大学 | Bifidobacterium longum capable of eliminating nonyl phenol and relieving poisoning symptoms caused by nonyl phenol |
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| CN111073828A (en) * | 2019-11-19 | 2020-04-28 | 江南大学 | Bifidobacterium longum subspecies longum and application thereof |
| CN113068837A (en) * | 2021-04-06 | 2021-07-06 | 江南大学 | Bifidobacterium longum capable of eliminating nonyl phenol and relieving poisoning symptoms caused by nonyl phenol |
Non-Patent Citations (2)
| Title |
|---|
| JINCHI JIANG等: "Strain-Specific Effects of Bifidobacterium longum on Hypercholesterolemic Rats and Potential Mechanisms" * |
| 姜金池: "益生菌对高胆固醇血症的缓解作用及机制研究" * |
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