CN110892961B

CN110892961B - Solid beverage containing bifidobacterium lactis BL-99 and application thereof

Info

Publication number: CN110892961B
Application number: CN201910567829.2A
Authority: CN
Inventors: 刘伟贤; 洪维鍊; 赵雯; 何剑; 陈醇
Original assignee: Inner Mongolia Yili Industrial Group Co Ltd
Current assignee: Inner Mongolia Yili Industrial Group Co Ltd
Priority date: 2019-06-27
Filing date: 2019-06-27
Publication date: 2022-03-29
Anticipated expiration: 2039-06-27
Also published as: CN110892961A

Abstract

The invention discloses a solid beverage containing bifidobacterium lactis BL-99 and application thereof. Specifically, the invention provides a solid beverage composition, which comprises the following raw materials by weight percent based on 100 percent of the total weight of the solid beverage: bifidobacterium lactis (Bifidobacterium lactis) with the preservation number of CGMCC No.15650 or 0.1-50% of active substances thereof; 10 to 20 percent of maltodextrin. The solid beverage has the effects of promoting intestinal digestion and relaxing bowels.

Description

Solid beverage containing bifidobacterium lactis BL-99 and application thereof

Technical Field

The invention relates to a solid beverage containing probiotics and application thereof, in particular to a solid beverage containing bifidobacterium lactis (BL-99) and application thereof in the aspects of promoting intestinal digestion, relaxing the bowels and the like.

Background

Today, with the pace of life increasing, irregular diets, frequent business trips, inadequate sleep, and inevitable alcohol consumption severely damage the digestive system of the body. While the imbalance of the digestive system is more likely to cause a series of intestinal problems: constipation, diarrhea, abdominal pain, borborborygmus, and … … air discharge, but if the intestinal peristalsis is slowed, the feces are retained in the intestinal tract and cannot be discharged in time, and harmful substances are generated in the feces in the intestinal tract, which seriously affects the human health. The results of the survey showed that 37.4% of 790 asian women suffered from constipation.

Researches show that the ingestion of probiotics can shorten the colon carrying time and simultaneously has certain help to regulate intestinal flora and improve gastrointestinal tract disorder of a subject. While colonic transit time may be defined as the time required for food to pass through swallowing, digestion, absorption and excretion. The health and morbidity of the gastrointestinal tract is sometimes associated with colonic transit time. Slow colonic transit represents symptoms such as constipation.

Disclosure of Invention

The invention discovers that Bifidobacterium lactis (Bifidobacterium lactis) BL-99 has the effect of promoting intestinal digestion, and accordingly provides a solid beverage containing the Bifidobacterium lactis BL-99.

According to the specific embodiment of the present invention, the Bifidobacterium lactis BL-99 of the present invention has been deposited in 26.04.2018 in the china general microbiological culture collection center CGMCC (address: beijing, north cheng yang district, west way No.1 institute of microbiology, china academy of sciences), and is named after classification: bifidobacterium lactis (Bifidobacterium lactis); the preservation number is CGMCC No. 15650. Bifidobacterium lactis (BL-99) has gastric acid and intestinal juice resistance, and has a viable bacteria survival rate of more than 62% when treated in gastric acid solution with pH of 2.5 for 30min and a viable bacteria survival rate of more than 61% when treated for 2 hours; the survival rate of viable bacteria is more than 70 percent after the intestinal juice with pH6.8 is treated for 2 hours.

The study of the invention finds that a single strain of bifidobacterium lactis BL-99 (namely bifidobacterium lactis with the preservation number of CGMCC No. 15650) has the effect of promoting intestinal digestion, can promote intestinal peristalsis and increase the output of pepsin.

Further, the invention provides a solid beverage composition, which comprises the following raw materials by weight percentage of the total weight of the solid beverage as 100 percent:

bifidobacterium lactis (Bifidobacterium lactis) with the preservation number of CGMCC No.15650 or 0.1-50% of active substances thereof;

10 to 20 percent of maltodextrin.

According to a specific embodiment of the present invention, in the solid beverage composition of the present invention, the active substance is a fermentation culture product of bifidobacterium lactis with a preservation number of CGMCC No.15650, including but not limited to: a fermentation culture containing bacterial cells, a fermentation supernatant from which the bacterial cells have been removed, or an active substance extracted from the fermentation culture. Bifidobacterium lactis BL-99 of the present invention can be cultured by anaerobic fermentation in a culture medium for Bifidobacterium lactis (e.g., TPY medium, BBL medium, etc.) which is commonly used in the art. The optimal fermentation temperature is 35-38 ℃, and the optimal fermentation time is 7-24 h. The invention also provides a preparation method of the fermentation culture product of the bifidobacterium lactis BL-99, which comprises the step of carrying out anaerobic culture on the strain in a liquid fermentation culture medium to obtain a fermentation liquid containing the strain. The fermentation liquor can be directly used as a liquid bacterial preparation or further concentrated, and the fermentation liquor can be dried to prepare bacterial powder, or the bacterial powder is prepared by separating thalli from the fermentation liquor. The liquid bacterial preparation of the present invention may be a liquid bacterial preparation prepared by suspending the bacterial cells in a culture medium, a buffer solution, deionized water or other solvent. The BL-99 liquid bacterium preparation or the solid bacterium preparation (bacterium powder) can be stored in a viable bacterium form and has better stability in a storage period. The BL-99 liquid bacteria preparation or solid bacteria preparation (bacteria powder) of the present invention can also be preserved in the form of inactivated dead bacteria. The bacterial preparation can be used for preparing the solid beverage. According to a preferred embodiment of the invention, the bifidobacterium lactis having a accession number CGMCC No.15650 is present in the solid beverage in the form of a viable organism.

According to a specific embodiment of the invention, in the solid beverage composition of the invention, the content of the bifidobacterium lactis with the preservation number of CGMCC No.15650 in the beverage composition is 1.0 x 10³CFU～1.0×10¹¹CFU/day, or 0.001 μ g-100 mg/day in terms of the weight of the cells.

According to a preferred embodiment of the present invention, in the solid beverage composition of the present invention, the content of bifidobacterium lactis having a collection number of CGMCC No.15650 in the solid beverage is 10⁸CFU～10¹⁰CFU/g, or 0.01 mu g-10 mg/day of thallus weight.

According to an embodiment of the present invention, maltodextrin can be used to help maintain the viable count of the product during shelf life in the solid beverage composition of the present invention.

According to a specific embodiment of the present invention, some conventional components of the existing probiotic solid beverage can be further included in the solid beverage composition of the present invention. In some embodiments of the present invention, the raw material composition of the solid beverage of the present invention further comprises one or more of fructo-oligosaccharide, xylo-oligosaccharide, erythritol.

According to a particular embodiment of the invention, the solid beverage composition of the invention further comprises a balance of fruit powder.

In some embodiments of the present invention, the solid beverage composition of the present invention comprises the following raw materials, based on the total weight of the solid beverage as 100%:

0.1-50% of bifidobacterium lactis or active substances thereof with the preservation number of CGMCC No. 15650;

10 to 20 percent of maltodextrin;

5 to 50 percent of fructo-oligosaccharide;

5 to 20 percent of xylo-oligosaccharide;

5% -20% of erythritol;

and (4) the balance of fruit powder.

In some embodiments of the present invention, in the solid beverage composition of the present invention, the fruit powder is orange fruit powder, and the content of the fruit powder in the solid beverage composition is 5% to 20%.

The preparation method of the solid beverage can be carried out by referring to the existing preparation technology of the probiotic solid beverage. In general, the solid beverage of the present invention can be obtained by uniformly mixing the components of the composition.

In another aspect, the invention also provides the application of the solid beverage composition in preparing a solid beverage with digestion promoting and/or constipation relieving effects.

The solid beverage of the invention has the corresponding effect of promoting intestinal digestion due to the inclusion of the bifidobacterium lactis BL-99, and can promote intestinal peristalsis and increase the output of pepsin. The solid beverage has better defecation effect.

Drawings

FIG. 1 shows the effect of Bifidobacterium lactis BL-99 on the ink propulsion rate (%).

FIG. 2 shows the effect of Bifidobacterium lactis BL-99 on the gastric fluid volume in rats.

FIG. 3 shows the effect of Bifidobacterium lactis BL-99 on rat pepsin activity.

FIG. 4 shows the effect of Bifidobacterium lactis BL-99 on pepsin output in rats.

FIG. 5 is a schematic view of a fermentation process according to an embodiment of the present invention.

FIG. 6 shows the effect of BL-99 solid drink of the present invention on defecation and digestion (results of human test questionnaire).

Microbial preservation of the patent procedure:

bifidobacterium lactis BL-99 of the present invention:

the preservation date is as follows: 26/04/2018;

the preservation unit: china general microbiological culture Collection center (CGMCC);

the address of the depository: xilu No.1 Hospital No. 3, the institute of microbiology, China academy of sciences, Beijing, Chaoyang

The preservation number is: CGMCC No. 15650;

and (3) classification and naming: bifidobacterium lactis (Bifidobacterium lactis).

Detailed Description

For a more clear understanding of the technical features, objects and advantages of the present invention, reference is now made to the following detailed description taken in conjunction with the accompanying specific embodiments, and the technical solutions of the present invention are described, it being understood that these examples are intended to illustrate the present invention and are not intended to limit the scope of the present invention. In the examples, each raw reagent material is commercially available, and the experimental method without specifying the specific conditions is a conventional method and a conventional condition well known in the art, or a condition recommended by an instrument manufacturer.

Unless specifically defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the relevant art. Unless otherwise indicated, all numbers expressing quantities of ingredients, cell culture, processing conditions, and so forth used in the present disclosure are to be understood as being modified in all instances by the term "about". Accordingly, unless indicated to the contrary, the numerical parameters are approximations and may vary depending upon the desired properties sought to be obtained by the present invention. The term "at least" preceding a series of elements is to be understood as referring to each element in the series, unless otherwise indicated.

In various examples of the invention, experimental data are expressed as Mean ± s.e.m. data were counted using PRISM version 5.0(GraphPad, San Diego, CA, USA). Differences between groups were counted using a one-way ANOVA followed by Tukery's multiple compliance test. There were significant statistical differences at P < 0.05.

Example 1: bifidobacterium lactis BL-99 and performance measurement thereof

The bifidobacterium lactis BL-99 of the invention is obtained from Shanghai Bingzhui GmbH and is separated from the intestinal tract of the infant. The strain has been preserved in China general microbiological culture Collection center (CGMCC) (address: No. 3 Xilu-Beijing province No.1, Beijing Korean district, Ministry of China microbiology institute) 26.04.2018, and is named after classification: bifidobacterium lactis (Bifidobacterium lactis); the preservation number is CGMCC No. 15650.

1. Taxonomical characterization of Bifidobacterium lactis BL-99

The results of the physical and chemical tests are as follows:

16S rRNA Gene sequence sequencing results (SEQ ID No. 1):

GCTCCCCCACAAGGGTCGGGCCACCGGCTTCGGGTGCTACCCACTTTCATGACTTGACGGGCGGTGTGTACAAGGCCCGGGAACGCATTCACCGCGGCGTTGCTGATCCGCGATTACTAGCGACTCCGCCTTCACGCAGTCGAGTTGCAGACTGCGATCCGAACTGAGACCGGTTTTCAGCGATCCGCCCCACGTCACCGTGTCGCACCGCGTTGTACCGGCCATTGTAGCATGCGTGAAGCCCTGGACGTAAGGGGCATGATGATCTGACGTCATCCCCACCTTCCTCCGAGTTGACCCCGGCGGTCCCACATGAGTTCCCGGCATCACCCGCTGGCAACATGCGGCGAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACGACCATGCACCACCTGTGAACCGGCCCCGAAGGGAAACCGTGTCTCCACGGCGATCCGGCACATGTCAAGCCCAGGTAAGGTTCTTCGCGTTGCATCGAATTAATCCGCATGCTCCGCCGCTTGTGCGGGCCCCCGTCAATTTCTTTGAGTTTTAGCCTTGCGGCCGTACTCCCCAGGCGGGATGCTTAACGCGTTGGCTCCGACACGGGACCCGTGGAAAGGGCCCCACATCCAGCATCCACCGTTTACGGCGTGGACTACCAGGGTATCTAATCCTGTTCGCTCCCCACGCTTTCGCTCCTCAGCGTCAGTGACGGCCCAGAGACCTGCCTTCGCCATTGGTGTTCTTCCCGATATCTACACATTCCACCGTTACACCGGGAATTCCAGTCTCCCCTACCGCACTCCAGCCCGCCCGTACCCGGCGCAGATCCACCGTTAGGCGATGGACTTTCACACCGGACGCGACGAACCGCCTACGAGCCCTTTACGCCCAATAAATCCGGATAACGCTCGCACCCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGGTGCTTATTCGAACAATCCACTCAACACGGCCGAAACCGTGCCTTGCCCTTGAACAAAAGCGGTTTACAACCCGAAGGCCTCCATCCCGCACGCGGCGTCGCTGCATCAGGCTTGCGCCCATTGTGCAATATTCCCCACTGCTGCCTCCCGTAGGAGTCTGGGCCGTATCTCAGTCCCAATGTGGCCGGTCACCCTCTCAGGCCGGCTACCCGTCAACGCCTTGGTGGGCCATCACCCCGCCAACAAGCTGATAGGACGCGACCCCATCCCATGCCGCAAAAGCATTTCCCACCCCACCATGCGATGGAGCGGAGCATCCGGTATTACCACCCGTTTCCAGGAGCTATTCCGGTGCACAGGGCAGGTTGGTCACGCATTACTCACCCGTTCGCCACTCTCACCCCGACAGCAAGCTGCCAGGGATCCCGTTCGACT

2. tolerance of bifidobacterium lactis BL-99 to artificial gastric juice and intestinal juice

Bifidobacteria are genera that are generally not acid-fast. In this example, the tolerance of Bifidobacterium lactis BL-99 of the present invention to artificial gastric juice and intestinal juice was tested, and Bifidobacterium lactis BB-

For comparison.

The test method comprises the following steps: culturing Bifidobacterium lactis BL-99 strain in MRS liquid culture medium at 37 deg.C for 16 hr, centrifuging at 4 deg.C and 2500rpm for 10min, and collecting thallus.

Respectively culturing the strains to be tested in artificial gastric juice and artificial small intestine juice, processing at 37 ℃ for 0, 30min and 2h, and then performing viable count analysis to evaluate the acid resistance and intestinal juice resistance of the strains according to the survival rate. Survival rate (viable cell count after treatment/viable cell count at time 0) × 100%.

The survival rate detection result of the bacterial strain in artificial gastric acid (pH2.5) is shown in Table 1, the survival rate of the viable bacteria is 7.04% when BB-12 is treated in the artificial gastric acid (pH2.5) for 30min, and the survival rate of the viable bacteria is only 1.64% after 2 hours of treatment; the survival rate of the live bacteria of the bifidobacterium lactis BL-99 is 62.60 percent when the bifidobacterium lactis BL-99 is treated in artificial gastric acid (pH2.5) for 30min, and the survival rate of the live bacteria is 61.83 percent when the bifidobacterium lactis BL-99 is treated for 2 hours. The bifidobacterium lactis BL-99 disclosed by the invention has excellent gastric acid resistance and can smoothly pass through the stomach to reach the intestinal tract to play a probiotic role.

TABLE 1 survival rate of the strains in artificial gastric acid (pH2.5)

The survival rate of the strain in the artificial small intestine solution (pH6.8) is tested and shown in Table 2. The data show that the viable bacteria survival rate of BB-12 in artificial small intestine solution (pH6.8) for 2 hours is only 28.95%; the viable bacteria survival rate of the bifidobacterium lactis BL-99 is 70.23 percent when the bifidobacterium lactis BL-99 is treated in artificial gastric acid (pH2.5) for 2 hours. The bifidobacterium lactis BL-99 disclosed by the invention has excellent intestinal juice resistance and can survive and colonize in intestinal tracts.

TABLE 2 survival rate of the strains in artificial intestinal juice (pH6.8)

3. Toxicity experiment and safety detection of bifidobacterium lactis BL-99

Inoculating the bifidobacterium lactis BL-99 of the invention into a BBL liquid culture medium, carrying out anaerobic culture for 48 +/-2 hours at 36 +/-1 ℃, and counting the viable count of the bifidobacterium lactis BL-99 in the culture solution to be 3.7 multiplied by 10⁸cfu/mL, stock solutions and 5-fold concentrates of the cultures were continuously gavaged to 20.0mL/kg BW for 3 days and observed for 7 days. The experiment was performed with a control group of 5-fold concentrated solution and a stock solution of the medium.

The test result shows that: the BBL culture stock solution and 5-fold concentrated solution of Bifidobacterium lactis BL-99 had no statistical effect on the weight gain of mice (p > 0.05) compared with the respective control group, and no toxic reaction or death of the tested mice was observed.

The antibiotic sensitivity of the bifidobacterium lactis BL-99 is evaluated by adopting an SN/T1944-2007 method of determination of bacterial resistance in animals and products thereof. The evaluation results show that the bifidobacterium lactis BL-99 is sensitive to Ampicillin Ampicillin, penicillin G Penicillin G, Erythromycin Erythromycin, Chloramphenicol Chloramphenicol, Clindamycin Clindamycin, Vancomycin Vancomycin, Tetracycline and the like. Meets the requirements of European Food Safety Authority (European Food Safety Authority) on the evaluation specification of the resistance of the edible bacteria. The bifidobacterium lactis BL-99 does not contain exogenous antibiotic resistance genes and is safe to eat.

4. Small bowel movement test

98 healthy SPF-grade BABL/c mice, weighing 18-20g, were provided by Beijing Huafukang Biotech, Inc. Raising in animal houses of Chinese disease prevention and control center: maintaining the room temperature (25 + -2 deg.C), relative humidity (55 + -2)%, and lighting for 12h/12h, and freely eating and drinking. According to the weight, the feed is divided into 7 groups of 14 animals by using random numbers, the animals are numbered by picric acid, and the animals are fed by common feed in a conventional way, and the grouping arrangement is shown in table 3.

TABLE 3 grouping arrangement

Grouping	Test article	Human daily dose (CFU/day)
			Control group	PBS	--
Model set	PBS	--
			BL-99 very low dose group	BL-99 bifidobacterium lactis	1×10⁶
BL-99 low dose group	BL-99 bifidobacterium lactis	1×10⁷
			BL-99 medium dose group	BL-99 bifidobacterium lactis	1×10⁸
BL-99 high dose group	BL-99 bifidobacterium lactis	1×10⁹
			BL-99 very high dose group	BL-99 bifidobacterium lactis	1×10¹⁰

Formal experiments were started 3 days after acclimation.

The test sample is administrated in a gastric lavage mode, the gastric lavage is carried out once a day, the gastric lavage volume is 20ml/kg, the control group and the model group are administrated with PBS in a gastric lavage mode, and the intervention time is 30 days.

During the experimental period, the animals are normal in characteristic, good in mental and activity conditions, and free of any adverse reaction after the test substances are given every day. As can be seen from the body weight data in Table 4, there was no significant difference in body weight between the groups (p > 0.05) over the experimental period.

TABLE 4 Effect of Bifidobacterium lactis BL-99 on mouse body weight (g)

FIG. 1 shows the effect of Bifidobacterium lactis BL-99 on the ink propulsion rate (%). As shown in FIG. 1, the ink propulsion rate of the mice in the model group is significantly lower than that of the control group (p < 0.05) compared with that of the control group, which indicates that the consistent mouse intestinal peristalsis model is successfully established. Compared with the model group, the BL-99 probiotic extremely low dose group (1X 10)⁶CFU/day), low dose group (1 × 10)⁷CFU/day) and high dose group (1 × 10)⁸CFU/day) mice had significantly higher ink propulsion rates than the model control (p < 0.05), and the experimental results were positive. Indicating a very low dose group (1X 10)⁶CFU/day), low dose group (1 × 10)⁷CFU/day) and high dose group (1 × 10)⁸CFU/day) BL-99 can promote intestinal movement of mice.

BL-99 and rat digestive enzyme assay

221 healthy SPF-grade SD rats weighing 180-. Raising in animal houses of Chinese disease prevention and control center: maintaining the room temperature (25 + -2 deg.C), relative humidity (55 + -2)%, and lighting for 12h/12h, and freely eating and drinking. The animals were divided into 17 groups of 13 animals each, assigned a picric acid number, and fed with normal diet according to the weight, using random numbers, and arranged in groups as shown in Table 3. Formal experiments were started 3 days after acclimation.

After the rats were numbered in groups using picric acid, they were weighed at

weeks

0, 1, 2, 3, and 4 after the test substance was administered, and the gavage amount of the rats was calculated. Rat feed was weighed weekly and rat food intake calculated. The test sample is administrated in a gastric lavage mode, the gastric lavage is carried out once a day, the gastric lavage volume is 20ml/kg, the control group and the model group are administrated with PBS in a gastric lavage mode, and the intervention time is 30 days.

Adding fresh egg white into a 2.0mm glass tube, placing the glass tube in boiling water for 3-5 seconds until the egg white turns white, taking out the glass tube, absorbing water on the tube wall by absorbent paper, and selecting a bubble-free protein tube for later use. Protein tubes were freshly made before the experiment. Rats were given a final dry prognosis and fasted for 24 hours prior to digestive enzyme assay. The depilatory cream is applied to the abdomen of the rat and the hair of about 4x4cm below the sternum of the rat is removed. The rats were anesthetized by inhalation anesthesia, placed on a laboratory bench with the abdomen facing up, and the skin of the abdomen of the rats was disinfected with iodophors. The skin of the rat below the sternum is cut by sterile scissors about 1cm, the chest cavity and the stomach of the rat are exposed, the opening is about 1-2 cm, the joint of the stomach and the duodenum of the rat (namely the pylorus of the rat) is found by sterile forceps, and the pylorus of the rat is ligated by using an operation suture. After the pylorus was ligated for 3 hours, the rats were killed by intraperitoneal injection of pentobarbital sodium, dissected, and the stomach of the rat was removed and collected in a centrifuge tube, and the amount of gastric juice per unit time was measured.

Taking 1ml of gastric juice, putting the gastric juice into a 50ml triangular flask, adding 15ml of 0.05mol/l hydrochloric acid solution, shaking up, and putting two freshly prepared protein tubes. The bottle mouth was closed and incubated at 37 ℃ for 24 hours. Protease activity and pepsin output were calculated.

Pepsin activity units (μ/mL) ═ length average of clear sections of tetrapolar protein tubes²×16；

Pepsin output (μ/h) is pepsin activity x gastric fluid volume per hour.

5.1 weight gain, food intake and food availability

Results after intervention are shown in Table 5, BL-99 in dose group (1X 10)⁸CFU/day) weight total gain and total food intake were significantly lower than those of the control group (p < 0.05), and the other groups had no significant difference in the total gain, total food intake and total food utilization index.

TABLE 5 Effect of Bifidobacterium lactis BL-99 on mouse Total weight gain, Total food intake, Total food utilization (g)

5.2 digestive enzymes

The results are shown in FIG. 2, and compared with the control group, the BL-99 middle dose group (1X 10)⁸CFU/day), BL-99 high dose group (1X 10)⁹CFU/day) gastric juice level was significantly higher than control (p < 0.05).

As shown in FIG. 3, the pepsin activity of each experimental group was not significantly different (p > 0.05) compared with the control group.

As shown in FIG. 4, the BL-99 middle dose group (1X 10) was compared with the control group⁸CFU/day) pepsin output was significantly higher than in the control group (p < 0.05).

The results show that the medium and high dosage of BL-99 bifidobacterium lactis improves the gastrointestinal digestion capability of mice to a certain extent and promotes gastric secretion.

Example 2: solid beverage containing bifidobacterium lactis BL-99

Referring to the fermentation process flow shown in fig. 5, bifidobacterium lactis BL-99 (i.e. bifidobacterium lactis with the preservation number of CGMCC No. 15650) provided by the invention is subjected to anaerobic culture in a TPY liquid culture medium. TPY liquid Medium (g/L): 10.0 parts of hydrolyzed casein, 5.0 parts of soytone, 2.0 parts of yeast powder, 5.0 parts of glucose, 0.5 part of L-cysteine, 2.0 parts of dipotassium phosphate, 0.5 part of magnesium chloride, 0.25 part of zinc sulfate, 0.15 part of calcium chloride, 0.0001 part of ferric chloride, and 801.0 parts of Tween, wherein the pH value is 6.5 +/-0.1. The fermentation broth after primary and secondary amplification culture is centrifuged at 2500rpm for 10min at 4 ℃ to collect the thallus. Freeze drying the collected thallus to obtain BL-99 active bacteria powder, and storing at-18 deg.c below.

The effect of BL-99 active bacterial powder of this example on the defecation function of mice was examined. Animal experiments prove that BL-99 pure fungus powder is 10 in percentage⁶～10¹⁰There were no statistically significant differences in CFU/day for the first bowel movement time, number of bowel movements, weight of bowel movements (table 6). Combining the digestion test results in example 1, it is clear that BL-99 itself, although having a digestion promoting effect, does not have the ability to improve defecation.

TABLE 6 influence of BL-99 active bacteria powder on animal defecation function

The solid beverage of this example was formulated according to the following ingredients in table 7, solid beverage product specifications: 2000 mg/bar.

TABLE 7 ingredient table for solid beverage

Batching table	The gram quantity (mg) is required for each bag	Percent (%)
			BL-99 bacterial powder	172	8.6％
Maltodextrin	288	14.4％
			Fructo-oligosaccharide	800	40.0％
Orange fruit powder	200	10.0％
			Xylo-oligosaccharide	200	10.0％
Erythritol and its preparation method	340	17.0％
			Total of	2000	100.0％

The solid beverage of the present example was prepared by mixing the above ingredients uniformly. The viable count in shelf life is maintained at 100 hundred million/bag.

The BL-99 solid beverage of the embodiment is subjected to crowd eating trial, and the results of questionnaire after two months show that the crowd eating the BL-99 solid beverage has better defecation somatosensory (figure 6, the percentage data in the figure represents the percentage of somatosensory population). The BL-99 solid beverage can increase the body feeling of digestion, increase the defecation times and reduce the defecation time.

Sequence listing

<110> Inmunogu Yili industry group GmbH

<120> solid beverage containing bifidobacterium lactis BL-99 and application thereof

<130> GAI19CN2300

<160> 1

<170> SIPOSequenceListing 1.0

<210> 1

<211> 1398

<212> DNA

<213> Bifidobacterium lactis (Bifidobacterium lactis)

<400> 1

gctcccccac aagggtcggg ccaccggctt cgggtgctac ccactttcat gacttgacgg 60

gcggtgtgta caaggcccgg gaacgcattc accgcggcgt tgctgatccg cgattactag 120

cgactccgcc ttcacgcagt cgagttgcag actgcgatcc gaactgagac cggttttcag 180

cgatccgccc cacgtcaccg tgtcgcaccg cgttgtaccg gccattgtag catgcgtgaa 240

gccctggacg taaggggcat gatgatctga cgtcatcccc accttcctcc gagttgaccc 300

cggcggtccc acatgagttc ccggcatcac ccgctggcaa catgcggcga gggttgcgct 360

cgttgcggga cttaacccaa catctcacga cacgagctga cgacgaccat gcaccacctg 420

tgaaccggcc ccgaagggaa accgtgtctc cacggcgatc cggcacatgt caagcccagg 480

taaggttctt cgcgttgcat cgaattaatc cgcatgctcc gccgcttgtg cgggcccccg 540

tcaatttctt tgagttttag ccttgcggcc gtactcccca ggcgggatgc ttaacgcgtt 600

ggctccgaca cgggacccgt ggaaagggcc ccacatccag catccaccgt ttacggcgtg 660

gactaccagg gtatctaatc ctgttcgctc cccacgcttt cgctcctcag cgtcagtgac 720

ggcccagaga cctgccttcg ccattggtgt tcttcccgat atctacacat tccaccgtta 780

caccgggaat tccagtctcc cctaccgcac tccagcccgc ccgtacccgg cgcagatcca 840

ccgttaggcg atggactttc acaccggacg cgacgaaccg cctacgagcc ctttacgccc 900

aataaatccg gataacgctc gcaccctacg tattaccgcg gctgctggca cgtagttagc 960

cggtgcttat tcgaacaatc cactcaacac ggccgaaacc gtgccttgcc cttgaacaaa 1020

agcggtttac aacccgaagg cctccatccc gcacgcggcg tcgctgcatc aggcttgcgc 1080

ccattgtgca atattcccca ctgctgcctc ccgtaggagt ctgggccgta tctcagtccc 1140

aatgtggccg gtcaccctct caggccggct acccgtcaac gccttggtgg gccatcaccc 1200

cgccaacaag ctgataggac gcgaccccat cccatgccgc aaaagcattt cccaccccac 1260

catgcgatgg agcggagcat ccggtattac cacccgtttc caggagctat tccggtgcac 1320

agggcaggtt ggtcacgcat tactcacccg ttcgccactc tcaccccgac agcaagctgc 1380

cagggatccc gttcgact 1398

Claims

1. The solid beverage composition comprises the following raw materials by weight percent based on the total weight of the solid beverage as 100 percent:

bifidobacterium lactis with preservation number of CGMCC number 15650 (Bifidobacterium lactis) Or 0.1% -50% of active substances thereof;

10% -20% of maltodextrin.

2. The solid beverage composition according to claim 1, wherein the active substance is a fermentation culture product of Bifidobacterium lactis having a accession number of CGMCC number 15650.

3. The solid beverage composition according to claim 1, wherein the bifidobacterium lactis having a collection number of CGMCC number 15650 is contained in the beverage composition in an amount of 1.0 x 10³CFU~1.0×10¹¹CFU/day, or 0.001 μ g-100 mg/day in terms of the weight of the bacteria.

4. The solid beverage composition according to claim 1, wherein the content of bifidobacterium lactis having a preservation number of CGMCC number 15650 in the solid beverage is 10⁸CFU~10¹⁰CFU/g, or 0.01 mu g-10 mg/day of thallus weight.

5. The solid beverage composition according to claim 1, wherein the solid beverage further comprises one or more of fructo-oligosaccharide, xylo-oligosaccharide, and erythritol.

6. A solid beverage composition according to claim 1 or 5, wherein the solid beverage further comprises fruit powder as the rest.

7. A solid beverage composition according to claim 6, wherein the fruit powder is orange fruit powder and is present in the solid beverage composition in an amount of from 5% to 20%.

8. The solid beverage composition according to claim 1, which comprises the following raw materials by weight percentage based on the total weight of the solid beverage as 100 percent:

0.1-50% of bifidobacterium lactis or active substances thereof with the preservation number of CGMCC number 15650;

10% -20% of maltodextrin;

5% -50% of fructo-oligosaccharide;

5% -20% of xylo-oligosaccharide;

5% -20% of erythritol;

and (4) the balance of fruit powder.

9. A solid beverage composition according to claim 8, wherein the fruit powder is orange fruit powder and is present in the solid beverage composition in an amount of from 5% to 20%.

10. Use of the solid beverage composition according to any one of claims 1 to 9 for preparing a solid beverage having digestion promoting and/or laxative effects.

11. The use according to claim 10, wherein the bifidobacterium lactis having a accession number of CGMCC number 15650 is present in the solid beverage in the form of a viable organism.