CN111254088A - Bacillus coagulans strain and application thereof - Google Patents
Bacillus coagulans strain and application thereof Download PDFInfo
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- CN111254088A CN111254088A CN201911392790.1A CN201911392790A CN111254088A CN 111254088 A CN111254088 A CN 111254088A CN 201911392790 A CN201911392790 A CN 201911392790A CN 111254088 A CN111254088 A CN 111254088A
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/742—Spore-forming bacteria, e.g. Bacillus coagulans, Bacillus subtilis, clostridium or Lactobacillus sporogenes
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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Abstract
The invention provides a bacillus coagulans strain and application thereof. The Bacillus coagulans strain (Bacillus coagulans) has a preservation number of CGMCC No. 17374. It is applied to the field of microbial preparation. The strain screened by the invention has the characteristics of high yield of spores and high yield of acid; has obvious difference with other strains of the same genus, high spore formation rate, good heat resistance and high fermentation level; it is a probiotic, and has the effects of general lactobacillus maintaining intestinal balance, stimulating immunity, and improving health level. The capsule preparation prepared by taking the compound as the core component has good functions of treating and adjusting various intestinal discomfort symptoms.
Description
Technical Field
The invention relates to the field of microbiology, in particular to a bacillus coagulans strain and application thereof.
Background
The importance of the health of the intestine, known as the second brain of the human body, is self evident. The cognition of the intestinal function has the functions of immunity and detoxification which are ignored by most people except common digestion and nutrient absorption. The food intake of a person during life is about 70 tons, which is 1000 times the body weight, and is handled by the intestinal tract. The total length of the large intestine and the small intestine of an adult is about 6 to 7 meters, which is an important organ for nutrient absorption, and 99 percent of various nutrient substances required by the human body are absorbed from the intestinal tract. Furthermore, the intestinal tract is the largest immune organ of human body, and a plurality of immune cells such as intestinal dendritic cells and the like and a plurality of immune effectors such as mucus layers, epithelial antibacterial proteins and the like participate in the intestinal tract mucous membrane immune response to resist a large amount of ingested or in-vivo generated toxins and harmful substances. Meanwhile, the toxin expelling agent is also the largest toxin expelling organ of a human body, and more than 80% of toxins of the human body are discharged by intestinal tracts. Based on the role of intestinal health in the overall health of the human body, the World Gastroenterology Organization (WGO) established the "world intestinal health day" in 2005.
However, as socio-economic development in our country progresses, numerous social factors threaten the health of the intestinal tract-antibiotic abuse in children behind parental anxiety disorder, female laxative dependence with a thin as aesthetic, an increase in the incidence of irritable bowel syndrome (ibs) under stress of working and living, and the like. Research shows that 54.5% of population in China has intestinal tract problems, the prevalence rate of adult chronic constipation is 14% on average, female is higher than male and can reach 17.4%, and the incidence rate of defecation frequency reduction of women with chronic constipation in the childbearing age is obviously higher than that of middle-aged and old people. The improper use of excretion-promoting drugs aggravates constipation, increases intestinal cancer risk and intestinal dysfunction; the patients are easy to form laxative dependence without using the medicine at will according to the clinical rules. In addition to this, irritable bowel syndrome (Ibs) is more prevalent in modern lifestyles: the physiological response of the entire intestine to the stimulus is excessive or abnormal, such as abdominal pain, diarrhea, constipation or repeated alternation of diarrhea and constipation. The prevalence of Ibs in the human population is 11.2%, and nearly 68% of Ibs patients continue to present symptoms for more than 12 years, resulting in a decrease in quality of life, even with psychopsychological disturbances. Authoritative survey shows that the current health situation of the intestinal tract of children in China is poor, the incidence rate of diarrhea in 2 weeks is 7%, and the total incidence rate of constipation is 4.7%. Parents rely on the effect of antibiotics, so that the rate of antibiotics abuse of children is as high as 19.9%, and intestinal flora imbalance occurs after 64.4% of children use the antibiotics. The long-term intestinal tract is unhealthy, so that the incidence of malignant diseases is continuously high. Taking colorectal cancer as an example, statistics data in 2015 show that the incidence and mortality of colorectal cancer in China are on the 5 th level in all malignant tumors, and the incidence of colorectal cancer is remarkably increased. China has become the world with the largest number of new annual average colorectal cancer cases.
The occurrence of the above-mentioned intestinal problems, the imbalance of the intestinal flora is the main cause. The inner cavity of the human intestinal tract is full of villi and microvilli, and the total area of the whole expansion can reach 200 square meters. The huge surface area of the intestinal tract is covered with over 1000 kinds of intestinal bacteria with the number as high as 100 trillion, which is almost 10 times of the total number of human cells. The intestinal flora is divided into three types: beneficial bacteria (such as lactic acid bacteria, bifidobacteria and yeasts), neutrophiles with high quantity which change with the change of human condition, and harmful bacteria (such as clostridium perfringens) which are easy to cause diseases. These bacteria, together with the surrounding environment, constitute the intestinal micro-ecosystem. The intestinal microecological system is changed due to subjective and objective reasons such as poor sleep quality, high stress, lack of dietary fibers of vegetables and fruits, lack of motion, dependence on cathartics, excessive intake of anthraquinone substances, abuse of antibiotics and the like, the stability of an in vitro organ of an organism, namely an intestinal flora, is damaged, the quantity of beneficial bacteria is reduced, and the quantity of harmful bacteria is increased. The instability of intestinal flora directly leads to the damage of human health, and the long-term accumulation of the intestinal flora causes larger pathological changes of the organism and vicious circle.
The symptomatic medicine administration and the supplement of beneficial bacteria become quite important and popular research directions for intestinal health. In 2001, the united nations Food and Agriculture Organization (FAO) and World Health Organization (WHO) have together proposed the definition of probiotics: probiotics are living microorganisms that, when ingested in sufficient quantities, exert a beneficial health effect on the host. Representative strains thereof include: lactobacillus, bifidobacterium and bacillus coagulans. The lactobacillus and the bifidobacterium are the most recognized probiotics for the public at present, and are widely applied to food, animal feed, clinic and other directions. Because of the limited structure of the non-spore-producing bacteria, the characteristics of no high temperature resistance and poor resistance to the external environment of the lactobacillus and the bifidobacterium exist, and a large amount of living bacteria can be apoptotic after a short storage time under the normal temperature condition. The supplement of human body to probiotics is mostly taken from digestive tract, and the bifidobacteria and the lactobacilli reach intestinal tracts after being treated by the strong acid environment and digestive enzymes of stomach, and the survival rates are respectively only 5.2 percent and 5.5 percent. Therefore, even if the external environment of low-temperature storage is kept, the probiotic effect of the bifidobacteria and the lactobacilli is greatly reduced after the bifidobacteria and the lactobacilli are ingested by a human body, and the effect of package propaganda of various products cannot be achieved.
Andres et al performed toxicological safety assessments of bacillus coagulans GenedenBC30TM, leading to a conclusion of "safe for long-term human consumption". The U.S. Food and Drug Administration (FDA) and the European Union food safety agency (EFSA) both passed the safety Qualification (QPS) for Bacillus coagulans. Bacillus coagulans has been used in the pharmaceutical, food and feed additive industries, etc. in japan, the united states, europe, etc. The application of Bacillus coagulans TQ33 as a food additive for chicks and growing-finishing pigs in New Star veterinary drug factory in Tianjin City was also approved in the 372 nd publication by the Ministry of agriculture in China. In 2005, live bacillus coagulans tablets produced by the east sea pharmaceutical co-production of Qingdao also obtained the new drug certificate of the State food and drug administration.
The most different of the bacillus coagulans from bifidobacteria and lactobacilli is that the produced spores can survive in severe environments such as strong acid or strong alkali. Data show that the survival rate of the bacillus coagulans in vitro after being tested for 10 minutes at 100 ℃ is more than 96%. Studies have shown that Bacillus coagulans strains can survive extreme conditions such as high temperature, gastric acid, and bile acid, under which conditions commonly-consumed probiotics die in large numbers. After being taken orally, the bacillus coagulans can smoothly pass through multiple gates such as gastric acid, digestive enzyme and the like and enter intestinal tracts to settle. They are rapidly revived there and become viable. Scientific research also shows that the bacillus coagulans can still maintain enough activity in an anoxic environment, can well grow in the anoxic environment in the intestinal tracts of people or animals, and can fully convert carbohydrate substances in food into D-type or D, L-type lactic acid, and the lactic acid can effectively kill or inhibit harmful microorganisms in the intestinal tracts. The bacillus coagulans and products (including lactic acid, bacteriocin and the like) thereof after planting regulate the environment in intestinal tracts, inhibit the growth of pathogenic bacteria, promote the increase of other beneficial bacteria, drive the beneficial functions of neutral bacteria, improve the immunity of organisms, improve the health state, play the medicinal role of reducing the use or replacing antibiotics, and further effectively solve the long-term intestinal problems caused by cathartic dependence and antibiotic abuse.
Disclosure of Invention
The invention provides a bacillus coagulans strain and application thereof.
The technical scheme of the invention is realized as follows:
a Bacillus coagulans strain (Bacillus coagulans) with preservation number of CGMCC No. 17374.
The 16S rDNA sequence of the strain is shown as SEQ ID NO. 1.
Culturing the bacillus coagulans strain to obtain a fermentation product.
A method of producing a fermentation product comprising:
primary culture, secondary culture, tertiary amplification culture and solid fermentation.
The first-stage culture comprises the following steps:
inoculating the original strain to a first-stage slant culture medium, and culturing at 30-45 ℃ for 20-48 h;
the first-stage slant culture medium is a mixed solution (g/L) prepared according to the following proportion: 6.0-15.0 g/L of yeast powder, 6.0-15.0 g/L of peptone, 2.0-10.0 g/L of glucose, 5.0-10.0 g/L of sodium chloride, 1.0-5.0 g/L of dipotassium hydrogen phosphate, 0.1-3.0 g/L of manganese sulfate, and 6.6-7.2 of pH.
The secondary culture comprises the following steps:
inoculating the single colony of the first-stage cultured bacillus coagulans into a second-stage culture solution, and culturing for 20-48 h at 30-45 ℃ and 150-300 rpm/min under the condition that the liquid loading is 50-100 mL/200mL/300mL/500mL to obtain a second-stage bacillus coagulans seed solution;
the secondary liquid culture medium is a mixed solution (g/L) prepared according to the following proportion: 10.0-20.0 g/L of glucose, 1.0-5.0 g/L of sodium chloride, 3.0-12.0 g/L of yeast powder, 10.0-30.0 g/L of tryptone, 5.0-15.0 g/L of beef extract powder, 1.0-5.0 g/L of dipotassium hydrogen phosphate, 1.0-5.0 g/L of magnesium sulfate, 1.0-5.0 g/L of manganese sulfate, 0.1-2.0 g/L of calcium carbonate, 0.5-5.0 g/L of calcium carbonate, and pH 6.6-7.2.
The three-stage amplification culture comprises the following steps:
culturing the bacillus coagulans seed liquid subjected to secondary culture for 20-48 h on a culture medium at the temperature of 30-45 ℃ and the rpm/min of 160-300 according to the volume percentage of 0.4-20.0% to obtain a bacillus coagulans tertiary seed liquid;
the culture medium is the culture medium in the secondary culture of the synchronous step.
The solid fermentation comprises the following steps:
inoculating a culture medium into a solid fermentation culture medium according to the weight ratio of 5-15%, controlling the water content of the culture medium to be 49-58%, controlling the pH value to be 6.0-7.0, keeping the relative humidity of a koji room to be more than 85%, controlling the material temperature to be 30-45 ℃, and fermenting for 48-72 hours by adopting a temperature control mode of ventilation and constant temperature in the early stage (24-48 hours), sealing in the later stage (48-72 hours) and stopping heating; and after the fermentation is finished, putting the mixture into an environment at 4 ℃ for 4-24 hours. The spore rate reaches more than 85 percent, and the spore rate reaches 2.0 multiplied by 1010 CFU/g.
The solid fermentation medium is prepared according to the following proportion: 35.0-45.0% of bran, 2.0-10.0% of corn flour, 20.0-30.0% of soybean meal, 0.1-2.0% of glucose, 2.0-5.0% of sodium chloride, 0.2-1.0% of dipotassium phosphate, 0.1-0.5% of manganese sulfate, 0.5-2.0% of calcium carbonate, 0.5-1.0% of ammonium sulfate and pH 6.6-7.2.
A microbial preparation comprising said fermentation product.
The microbial preparation is a viable bacteria quantitative of 1.5 multiplied by 108～3.0×108CFU/capsule.
The microbial preparation is suitable for relieving and curing discomfort symptoms of intestinal tracts of human bodies: the discomfort symptoms comprise long-term or short-term constipation, laxative dependence type constipation, seasonal or common diarrhea, alcoholic colitis, abdominal distension and other intestinal disease symptoms.
A method for producing a microbial preparation, comprising the step of culturing the Bacillus coagulans strain of claim 1.
Advantageous effects
(1) The strain screened by the invention has the characteristics of high yield of spores and high yield of acid.
(2) The bacillus coagulans provided by the invention has obvious difference with other congeneric strains, and has the advantages of very high spore formation rate, good heat resistance and high fermentation level.
(3) The bacillus coagulans provided by the invention is a probiotic and has the effects of maintaining intestinal balance of common lactic acid bacteria, stimulating immunity, improving the health level of organisms and the like.
(4) The bacillus coagulans provided by the invention can be used as probiotics (microbial preparations) to be applied to relieving and curing human intestinal discomfort symptoms (intestinal disease symptoms such as long-term or short-term constipation, laxative dependence type constipation, seasonal or common diarrhea, alcoholic colitis, abdominal distension and the like).
(5) The bacillus coagulans provided by the invention can produce a large amount of L-lactic acid, and can produce 0.9ug/L L-lactic acid under anaerobic conditions.
(6) The invention adopts a solid fermentation process, a heterogeneous system is more beneficial to the reproduction of the bacillus coagulans and the formation of spores, and compared with the traditional liquid fermentation process, the spore rate is improved by 35-50%. Compared with the prior fermentation process of the company, the spore rate is improved by 5 to 10 percent. The invention adopts the solid fermentation process of controlling the air flow in the early and late stages, and combines the low-temperature stimulation to the fermentation product, thereby not only solving the problems of insufficient acid production and mixed bacteria pollution in the metabolic products, but also promoting the rapid formation of spores.
(7) The bacillus coagulans provided by the invention can be propagated in a culture medium with deficient nutrition, and the industrial residues with lower cost such as bran, bean pulp and the like are adopted, so that the cost is saved, the operation is simple and convenient, and a better product can be obtained.
(8) The bacillus coagulans provided by the invention has strong stress resistance due to the property of producing spores, and can avoid great loss of bacterial quantity in the processes of transportation, processing and storage.
(9) The microbial agent prepared by using the provided bacillus coagulans comprises bacillus coagulans thalli, spores and metabolites thereof in the fermentation process.
(10) The microbial agent capsule prepared by using the provided bacillus coagulans has feasible preparation method, simple process and easy operation, and improves the utilization value of the microbial agent.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without inventive exercise.
FIG. 1 is a graph comparing the effect of the administration of the compound to the subjects in example 3.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
And (3) biological preservation:
the Bacillus coagulans strain (Bacillus coagulans) is preserved in China general microbiological culture Collection center (CGMCC for short; address: China academy of sciences microorganism institute 3, No.1, Xilu, Beijing, the rising district, Beijing, and the postal code: 100101) in 20 days 3 months and 2019, and the preservation number is as follows: CGMCC NO. 17374.
And (3) biological identification:
1. morphological identification
(1) The diameter of a bacterial colony on the improved YPD agar culture medium is 2.0-3.0 mm, the bacterial colony is flat, the edge is neat, the bacterial colony is opaque, glossy and pigment-free, the surface is moist, and the texture is soft.
(2) Gram staining is carried out on the bacillus coagulans colony, morphological characteristics of the strain are observed through an oil mirror, the bacillus coagulans colony is found to be a gram-positive bacterium, the size of the bacterium is (0.6-1.0) Mumx (2.5-5.0) Mum, and the bacterium is rod-shaped, oval in spore shape and terminal growth.
2. Physiological and biochemical identification
Various biochemical identification tests such as growth tests, V.P. tests, different substrate digestion tests, acid production tests and the like under different conditions are carried out, and the results are shown in the following table 1.
TABLE 1 Biochemical identification of Bacillus coagulans
Note: + indicates positive results and-negative results.
Tests prove that the strain can grow under the conditions of anaerobism, 7% of sodium chloride and pH5.7, the V.P. test is positive, starch can be hydrolyzed, gelatin can be liquefied, propionate is utilized, acid is produced, and the characteristics of bacillus coagulans are met.
The strain can increase the bacterial quantity under the aerobic condition, and the total metabolism in the fermentation process generates a small amount of L-lactic acid; can produce spores under anaerobic condition and can produce more L-lactic acid.
Example 1
A Bacillus coagulans strain (Bacillus coagulans) is screened out by Hangzhou Baoankang biotechnology limited, the preservation number of the Bacillus coagulans strain is CGMCC No.17374, and the Bacillus coagulans strain is a new strain.
The fermentation process comprises the following steps:
(1) first-stage slant culture of bacillus coagulans:
inoculating the original strain to a first-stage slant culture medium, and culturing at 30-45 ℃ for 20-48 h. The first-stage slant culture medium is a mixed solution (g/L) prepared according to the following proportion: 6.0-15.0 g/L of yeast powder, 6.0-15.0 g/L of peptone, 2.0-10.0 g/L of glucose, 5.0-10.0 g/L of sodium chloride, 1.0-5.0 g/L of dipotassium hydrogen phosphate, 0.1-3.0 g/L of manganese sulfate, and 6.6-7.2 of pH
(2) Secondary culture of bacillus coagulans:
inoculating the single colony of the first-stage bacillus coagulans in the step (1) into a second-stage culture solution, wherein the liquid loading amount is 50-100 mL/200mL/300mL/500mL, and the second-stage bacillus coagulans seed solution is obtained by culturing for 20-48 h at 30-45 ℃ and 150-300 rpm/min.
The secondary liquid culture medium is a mixed solution (g/L) prepared according to the following proportion: 10.0-20.0 g/L of glucose, 1.0-5.0 g/L of sodium chloride, 3.0-12.0 g/L of yeast powder, 10.0-30.0 g/L of tryptone, 5.0-15.0 g/L of beef extract powder, 1.0-5.0 g/L of dipotassium hydrogen phosphate, 1.0-5.0 g/L of magnesium sulfate, 1.0-5.0 g/L of manganese sulfate, 0.1-2.0 g/L of calcium carbonate, 0.5-5.0 g/L of calcium carbonate, and pH 6.6-7.2.
(3) B, three-stage amplification culture of the bacillus coagulans:
and (3) culturing the second-stage bacillus coagulans seed liquid obtained in the step (2) for 20-48 h on a culture medium at the temperature of 30-45 ℃ and the rpm of 160-300/min to obtain third-stage bacillus coagulans seed liquid, wherein the volume percentage of the second-stage bacillus coagulans seed liquid is 0.4-20.0%.
The culture medium is synchronous with the culture medium in the step (2).
(4) Solid fermentation of bacillus coagulans:
the culture medium is inoculated into a solid fermentation culture medium according to the weight ratio of 5-15%, the water content of the culture medium is controlled at 49-58%, the pH range is 6.0-7.0, the relative humidity of a koji room is kept at more than 85%, the material temperature is controlled at 30-45 ℃, and fermentation is carried out for 48-72 hours by adopting a temperature control mode of ventilation and constant temperature in the early stage (24-48 hours), sealing in the later stage (48-72 hours) and stopping heating. And after the fermentation is finished, putting the mixture into an environment at 4 ℃ for 4-24 hours. The spore rate reaches more than 85 percent, and the spore rate reaches 2.0 multiplied by 1010CFU/g。
The solid fermentation medium is prepared according to the following proportion: 35.0-45.0% of bran, 2.0-10.0% of corn flour, 20.0-30.0% of soybean meal, 0.1-2.0% of glucose, 2.0-5.0% of sodium chloride, 0.2-1.0% of dipotassium phosphate, 0.1-0.5% of manganese sulfate, 0.5-2.0% of calcium carbonate, 0.5-1.0% of ammonium sulfate and pH 6.6-7.2.
The bacillus coagulans is cultured by the existing solid fermentation production technology to obtain a bacillus coagulans culture with high density and high sporulation rate. After solid fermentation and culture for 48 hours, the number of viable bacteria reaches 1.0 multiplied by 1010CFU/g, the number of spores can reach 5 multiplied by 109CFU/g, the spore rate is only about 50% and needs to be further improved. Further experiments have shown that the adjustment of the culture conditions can optimize the spore transformation rate of Bacillus coagulans. The culture medium comprises the following components: the carbon source is sufficient to facilitate the growth of the thalli, and the lack of the carbon source can promote the formation of spores; in terms of culture temperature: the low temperature is easier to form spores than the medium and high temperature; the aspect of breathing energy consumption is as follows: spores are more easily generated under anaerobic and facultative anaerobic conditions. The method adopts a nutritional formula with high carbon source and nitrogen source to promote the growth of the bacteria, and carries out low-temperature treatment after the bacterial quantity is increased and enters a stable period, thereby obtaining the effective preparation method of the high-quality bacillus coagulans product.
Example 2
The Bacillus coagulans strain (Bacillus coagulans) has the preservation number of CGMCC No.17374, and is applied in the direction of microbial inoculum.
The method comprises the following specific steps:
(1) and (3) collecting and drying the solid fermentation product of the bacillus coagulans in the embodiment 1, wherein the drying temperature is 40-60 ℃, the drying time is 18-36 hours, and the drying is stopped until the water content is 5-10%.
(2) And (2) crushing the dried material in the step (1), sieving the particles with a 60-mesh sieve, collecting and bagging, and sealing and storing in a cool and dry place.
(3) And (3) measuring the live bacterial quantity and the spore number of the bacillus coagulans of the microbial agent in the step (2).
(4) Based on the measurement result in step (3), the step (c) is performedThe powder in the step (2) is quantitatively filled into a capsule shell, and the living bacteria quantitative is 1.5 multiplied by 108～3.0×108CFU/capsule.
The capsule is a commercially available medicinal capsule shell.
The method for packaging and sealing is not particularly limited, and the conventional packaging and sealing method in the field can be adopted.
Example 3
The microbial preparation capsules described in example 2 were tested for bioefficacy.
First, 50 subject volunteers volunteered to participate in the study and signed an informed consent.
Grouping standard:
inclusion criteria were: (1) has symptoms of intestinal diseases such as long-term or short-term constipation, constipation due to drug dependence, seasonal or common diarrhea, alcoholic colitis, and abdominal distention; (2) age 8 to 60 years; (3) voluntarily attend the study and sign informed consent.
Second, method
1. The research method comprises the following steps: the volunteers were examined for symptoms of intestinal diseases, 50 persons who met the entry criteria were selected and given the bacillus coagulans microbial preparation capsules described in application example 1, and a 60-day comparative test was carried out and the effect of continuous care was continued after discontinuation of the administration.
2. Evaluation basis: volunteers self-describe, and test intestinal disease symptoms.
3. And (3) intervention flow:
the subject takes 1 capsule once twice a day within 30min after meal.
4. The statistical method comprises the following steps: statistical analysis was performed using SPSS 22.0. The difference is statistically significant when P is less than 0.05.
Three, result in
The results before and after administration are shown in FIG. 1. The comparison of the number of days for which the symptoms of the intestinal disease were relieved and cured is shown in table 1.
The results in figure 1 show that after 60 days of administration, the administration effect of the subject is obvious, the number of effective people is 49 people, the effective rate is 98.0%, and the difference between groups is obvious (P is less than 0.05). The recurrence rate is 6.0%, i.e. the administration is stopped after 60 days, and the intestinal disease symptoms continue to appear. The disease recurrence phenomenon after the microbial preparation capsule is stopped is presumed to occur in the subjects with short colonization time and poor intestinal tract self-repairing capability due to different intestinal tract colonization time of the bacillus coagulans in the intestinal tract and the difference of the intestinal tract repairing capability of the subjects.
As can be seen from Table 1, the time for the Bacillus coagulans microbial agent capsule to relieve or cure the intestinal diseases is prolonged as the intestinal diseases become more severe. This result also laterally reflects the length of time that intestinal function is restored after bacillus coagulans enters the intestine to begin regulatory function.
TABLE 1 comparison of days for different degrees of symptomatic relief or cure of intestinal disorders
And selecting volunteers to perform a functional test on the premise of complete volunteers. The test result is good, the effect of the preparation is consistent with that of the bacillus coagulans clinically reported in the literature, and the preparation effect is superior to the effect of the bacillus coagulans and the antibiotic or the montmorillonite powder which are combined for treating diarrhea in the prior report. The medicine has obvious improvement on symptoms such as alcoholic colitis, short-term or long-term constipation, drug dependence constipation, seasonal or common diarrhea, abdominal distension and the like, has no side effect symptom in the whole population and has low recurrence rate.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Sequence listing
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<120> bacillus coagulans strain and application thereof
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tcaggacgaa cgctggcggc gtgcctaata catgcaagtc gtgcggacct tttaaaagct 60
tgcttttaaa aggttagcgg cggacgggtg agtaacacgt gggcaacctg cctgtaagat 120
cgggataacg ccgggaaacc ggggctaata ccggatagtt ttttcctccg catggaggaa 180
aaaggaaaga cggctttggc tgtcacttac agatgggccc gcggcgcatt agctagttgg 240
tggggtaacg gctcaccaag gcaacgatgc gtagccgacc tgagagggtg atcggccaca 300
ttgggactga gacacggccc aaactcctac gggaggcagc agtagggaat cttccgcaat 360
ggacgaaagt ctgacggagc aacgccgcgt gagtgaagaa ggccttcggg tcgtaaaact 420
ctgttgccgg ggaagaacaa gtgccgttcg aacagggcgg cgccttgacg gtacccggcc 480
agaaagccac ggctaactac gtgccagcag ccgcggtaat acgtaggtgg caagcgttgt 540
ccggaattat tgggcgtaaa gcgcgcgcag gcggcttctt aagtctgatg tgaaatcttg 600
cggctcaacc gcaagcggtc attggaaact gggaggcttg agtgcagaag aggagagtgg 660
aattccacgt gtagcggtga aatgcgtaga gatgtggagg aacaccagtg gcgaaggcgg 720
ctctctggtc tgtaactgac gctgaggcgc gaaagcgtgg ggagcaaaca ggattagata 780
ccctggtagt ccacgccgta aacgatgagt gctaagtgtt agagggtttc cgccctttag 840
tgctgcagct aacgcattaa gcactccgcc tggggagtac ggccgcaagg ctgaaactca 900
aaggaattga cgggggcccg cacaagcggt ggagcatgtg gtttaattcg aagcaacgcg 960
aagaacctta ccaggtcttg acatcctctg acctccctgg agacagggcc ttccccttcg 1020
ggggacagag tgacaggtgg tgcatggttg tcgtcagctc gtgtcgtgag atgttgggtt 1080
aagtcccgca acgagcgcaa cccttgacct tagttgccag cattcagttg ggcactctaa 1140
ggtgactgcc ggtgacaaac cggaggaagg tggggatgac gtcaaatcat catgcccctt 1200
atgacctggg ctacacacgt gctacaatgg atggtacaaa gggctgcgag accgcgaggt 1260
taagccaatc ccagaaaacc attcccagtt cggattgcag gctgcaaccc gcctgcatga 1320
agccggaatc gctagtaatc gcggatcagc atgccgcggt gaatacgttc ccgggccttg 1380
tacacaccgc ccgtcacacc acgagagttt gtaacacccg aagtcggtga ggtaaccttt 1440
acggagccag ccgccgaagg tgggacagat gattggggtg aagtc 1485
Claims (8)
1. A Bacillus coagulans strain (Bacillus coagulans) with preservation number of CGMCC No. 17374.
2. The strain of claim 1, wherein the 16S rDNA sequence is set forth in SEQ ID NO 1.
3. A fermentation product obtained by culturing the Bacillus coagulans strain of claim 1.
4. A method of producing a fermentation product of claim 3, comprising:
primary culture, secondary culture, tertiary amplification culture and solid fermentation.
5. A microbial preparation comprising the fermentation product of claim 3.
6. The microbial preparation of claim 5, wherein the microbial preparation is viable bacteria in an amount of 1.5 x 108～3.0×108CFU/capsule.
7. The microbial preparation according to claim 5, wherein said microbial preparation is suitable for the alleviation and cure of intestinal discomfort symptoms in humans: the discomfort symptoms are intestinal disease symptoms including long-term or short-term constipation, laxative-dependent constipation, seasonal or common diarrhea, alcoholic colitis, and abdominal distension.
8. A process for producing a microbial preparation, which comprises the step of culturing the Bacillus coagulans strain of claim 1.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
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| CN113198004A (en) * | 2021-05-19 | 2021-08-03 | 天津市宝恒生物科技有限公司 | Bacillus coagulans preparation for repairing intestinal mucosa |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112941005A (en) * | 2021-01-22 | 2021-06-11 | 武汉微康益生菌研究院有限公司 | Method for rapidly producing spores by bacillus coagulans |
| CN113198004A (en) * | 2021-05-19 | 2021-08-03 | 天津市宝恒生物科技有限公司 | Bacillus coagulans preparation for repairing intestinal mucosa |
| CN114480218A (en) * | 2022-03-14 | 2022-05-13 | 善恩康生物科技（苏州）有限公司 | Bacillus coagulans for improving constipation and application thereof |
| CN116925960A (en) * | 2023-06-05 | 2023-10-24 | 山东弥美生物科技股份有限公司 | Wettman-coagulating strain capable of utilizing pullulan and application thereof |
| CN116925960B (en) * | 2023-06-05 | 2024-01-05 | 山东弥美生物科技股份有限公司 | A strain of Weizmannella coagulans capable of utilizing pullulan and its application |
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