CN115353988B - Lactobacillus paracasei LC-37 with digestion promoting effect and application - Google Patents
Lactobacillus paracasei LC-37 with digestion promoting effect and application Download PDFInfo
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- CN115353988B CN115353988B CN202210574483.0A CN202210574483A CN115353988B CN 115353988 B CN115353988 B CN 115353988B CN 202210574483 A CN202210574483 A CN 202210574483A CN 115353988 B CN115353988 B CN 115353988B
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Abstract
The invention belongs to the field of microbial fermentation, and in particular relates to lactobacillus paracasei (Lactobacillus paracasei) LC-37 with digestion promoting effect and application thereof, and provides lactobacillus paracasei (Lactobacillus paracasei) LC-37 which is preserved in China general microbiological culture collection center (CGMCC) with a preservation number of CGMCC No.14055. The lactobacillus paracasei LC-37 is separated from intestinal contents of old people with health and longevity of Guangxi Bama, has the effect of promoting digestion, is particularly used for improving dyspepsia symptoms such as abdominal pain, belch, pantothenic acid, abdominal distention, inappetence, diarrhea or constipation of dyspepsia people, and can increase the content of short-chain fatty acid in the intestinal tract.
Description
Technical Field
The invention belongs to the field of microbial fermentation, and particularly relates to lactobacillus paracasei (Lactobacillus paracasei) LC-37 with digestion promoting effect and application thereof.
Background
Lactobacillus paracasei (Lactobacillus paracasei) is one of the populations of Lactobacillus, an important probiotic, and is commonly used in the production of dairy products, health foods, beverages, ice cream and the like. Numerous studies have shown that lactobacillus paracasei has the functions of regulating intestinal flora, enhancing immunity and the like. A Lactobacillus paracasei ET-22 with constipation relieving function is disclosed in Chinese patent document CN110892989A, wherein the Lactobacillus paracasei ET-22 with constipation relieving function is introduced, and the main function of the strain is to improve the intestinal propulsion rate and shorten the operation time of the intestinal tract, thereby playing a role in relieving constipation. And a new application of the lactobacillus paracasei K56 disclosed in the Chinese patent document CN110882280A, wherein the lactobacillus paracasei CGMCC15139 has the effect of obviously relieving and/or preventing constipation, and can improve the intestinal canal propulsion rate and shorten the defecation time.
With the development of society, the life rhythm is continuously accelerated, the working pressure is increased, and the dyspepsia problem caused by the problems of on-time meal, fast food and the like cannot be guaranteed is gradually highlighted. Eating foods with digestion promoting effects is an important and convenient means for improving the problem of dyspepsia. However, there are few studies on the digestion promoting effect of lactobacillus paracasei at present, and there is no effective evaluation of the digestion-related function.
Disclosure of Invention
Therefore, the technical problem to be solved by the invention is to provide lactobacillus paracasei (Lactobacillus paracasei) LC-37 with digestion promoting effect and application.
The invention provides a lactobacillus paracasei (Lactobacillus paracasei) strain LC-37 which is preserved in China general microbiological culture Collection center (CGMCC), and has the preservation address: beijing, chaoyang, north Chen Xi Lu 1, 3, china academy of sciences microbiological institute, postal code: 100101, and the preservation number is CGMCC No.14055.
The invention provides a microbial preparation, which takes lactobacillus paracasei LC-37, an inactivated product of lactobacillus paracasei LC-37 or a treated product of lactobacillus paracasei LC-37 as an active ingredient;
the treated product of the lactobacillus paracasei LC-37 is at least one of a culture, a crushed product and an extract of the culture of the lactobacillus paracasei LC-37.
The microbial preparation further comprises at least one excipient suitable for the microbial preparation; the microbial preparation is powder, pill, capsule, granule, tablet, liquid preparation or gel.
The invention provides a preparation method of lactobacillus paracasei LC-37 bacterial powder, which is characterized in that fermentation liquor of lactobacillus paracasei LC-37 is centrifuged, bacterial cells are collected, a freeze-drying protective agent is added into the obtained bacterial cells, and freeze-dried powder is obtained through vacuum freeze-drying.
In the preparation method, the fermentation steps of the lactobacillus paracasei LC-37 are as follows: inoculating seed liquid of lactobacillus paracasei LC-37 into a fermentation culture medium with an inoculum size of 2% -5% (v/v), culturing at 33-40 ℃, stirring at 100-200 rpm, maintaining pH of the culture medium at 5.0-6.5, and fermenting for 14-20 hours.
In the preparation method, the centrifugation conditions are as follows: the rotating speed is 8000-10000 rpm, and the centrifugal force is 1-2 hours.
In the preparation method, the freeze-drying protective agent comprises the following raw materials in parts by weight: 1 to 3 parts of trehalose, 1 to 3 parts of skim milk powder, 2 to 4 parts of maltodextrin and 0.1 to 0.5 part of sodium ascorbate.
The mass ratio of the addition of the freeze-drying protective agent to the thalli obtained by centrifugation is 0.2-0.9:1, the temperature is increased to 30 ℃ within 40-60 hours under the condition of vacuum freeze drying, and the vacuum degree is 3-50 Pa.
The preparation method further comprises the steps of: inoculating lactobacillus paracasei LC-37 frozen strain into 10-50 mL culture medium, and culturing at 33-40 ℃ for 12-24 hours.
In the preparation method, the freeze-dried powder is further added with auxiliary materials, wherein the auxiliary materials can be maltodextrin, whey powder, fructo-oligosaccharide or resistant starch.
The invention provides lactobacillus paracasei LC-37 powder prepared by the preparation method of the lactobacillus paracasei LC-37 powder.
The invention provides lactobacillus paracasei LC-37, the microbial preparation, the bacterial powder prepared by the method or the application of the bacterial powder in preparing medicines or foods with digestion promoting effect.
In the present invention, the medicine may be prepared in different forms according to the administration route, for example, may be prepared in the form of powder, tablet, granule, capsule, solution, emulsion, suspension, etc.; pharmaceutically acceptable adjuvants may also be included in the medicament. Food products include any type of food product, such as solid beverages, soy products, fruit juices, dairy products, ice creams, confectioneries, biscuits, and the like; the food may also contain conventional additives, nutrition enhancer, and adjuvants, such as essence, flavoring agent, stabilizer, thickener, antiseptic, mineral, vitamins, maltodextrin, etc.
The invention provides a preparation method of lactobacillus paracasei LC-37-containing lactobacillus paracasei beverage, which comprises the steps of using the lactobacillus paracasei LC-37, the microbial preparation, the bacterial powder prepared by the method or the bacterial powder.
In the preparation method, lactobacillus paracasei LC-37 strain powder is inoculated in the fermentation step of the drink base material, and the inoculation amount is 10 4 -10 7 CFU/mL, fermentation temperature is 33-40 ℃, and fermentation time is 72-100 hours.
In the preparation method, the viable count range of the lactobacillus paracasei LC-37 in the prepared lactobacillus beverage is more than or equal to 5 multiplied by 10 7 CFU/mL。
In the preparation method, the beverage base material comprises the following raw materials in parts by weight: 110-140 parts of skim milk powder, 15-25 parts of glucose and 35-50 parts of white granulated sugar.
In the preparation method, the beverage base material is subjected to sterilization and browning before fermentation, wherein the sterilization and browning conditions are 90-97 ℃ and 2-5 hours.
In the preparation method, the method further comprises the step of adding auxiliary materials into the fermented milk obtained in the fermentation step. The auxiliary materials can be sugar solution and/or essence, etc.
The invention provides a lactobacillus beverage containing lactobacillus paracasei LC-37 prepared by the method.
Compared with the prior art, the invention has the following advantages:
1. the invention provides lactobacillus paracasei (Lactobacillus paracasei) LC-37 which is preserved in the China general microbiological culture Collection center with the preservation number of CGMCC No.14055. The lactobacillus paracasei LC-37 is separated from intestinal contents of old people with good health and long life of Guangxi Bama, has the effect of improving dyspepsia, and is particularly used for improving dyspepsia symptoms such as abdominal pain, belch, pantothenic acid, abdominal distention, inappetence, diarrhea or constipation of dyspepsia people, and can increase the content of short chain fatty acid in the intestinal tract.
2. The lactobacillus beverage containing the lactobacillus paracasei LC-37 provided by the invention utilizes the lactobacillus paracasei LC-37 as a fermentation strain, and the prepared lactobacillus beverage has the effect of promoting digestion, can improve the symptoms of dyspepsia such as abdominal pain, belch, pantothenic acid, abdominal distention, inappetence, diarrhea or constipation and the like, and can increase the content of short-chain fatty acid in intestinal tracts.
3. The lactobacillus paracasei LC-37 powder provided by the invention has the effect of promoting digestion, can improve the symptoms of dyspepsia such as abdominal pain, belch, pantothenic acid, abdominal distention, inappetence, diarrhea or constipation and the like, and can increase the content of short-chain fatty acid in intestinal tracts.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the description of the embodiments or the prior art will be briefly described, and it is obvious that the drawings in the description below are some embodiments of the present invention, and other drawings can be obtained according to the drawings without inventive effort for a person skilled in the art.
FIG. 1 is a graph showing the change in scores of abdominal pain symptoms in subjects in the experimental example of the present invention;
FIG. 2 shows the change in scores of the eructation symptoms of the subjects in the experimental example of the present invention;
FIG. 3 is a graph showing the change in scores of pantothenic acid symptoms in subjects in the experimental example of the present invention;
FIG. 4 shows the change in scores of abdominal distension symptoms in subjects in the experimental example of the present invention;
FIG. 5 shows the change in scores of appetite symptoms in subjects in the experimental example of the present invention;
FIG. 6 shows the change in scores of diarrhea and diarrhea symptoms in subjects in the experimental example of the present invention;
FIG. 7 is a change in the integral of total clinical symptoms of dyspepsia in subjects in experimental examples of the invention;
FIG. 8 is a graph showing the change in acetic acid content before and after drinking by a subject in an experimental example of the present invention;
FIG. 9 is a graph showing the changes in propionic acid content before and after drinking by a subject in an experimental example of the present invention;
FIG. 10 is a graph showing the change in butyric acid content before and after drinking by a subject in an experimental example of the present invention;
FIG. 11 is a graph showing the change in total acid content before and after drinking by a subject in an experimental example of the present invention;
fig. 12 is a standard graph in experimental examples of the present invention.
Detailed Description
The following examples are provided for a better understanding of the present invention and are not limited to the preferred embodiments described herein, but are not intended to limit the scope of the invention, any product which is the same or similar to the present invention, whether in light of the present teachings or in combination with other prior art features, falls within the scope of the present invention.
The specific experimental procedures or conditions are not noted in the examples and may be followed by the operations or conditions of conventional experimental procedures described in the literature in this field. The reagents or apparatus used were conventional reagent products commercially available without the manufacturer's knowledge.
Embodiments of the present invention are described below by way of specific examples, wherein reagents and materials used in the examples are commercially available, using techniques conventional in the art, unless otherwise indicated.
EXAMPLE 1 acquisition of Lactobacillus paracasei strain LC-37
1. Isolation of Lactobacillus paracasei Strain LC-37
Collecting intestinal contents, namely faeces, of the Guangxi Bama healthy and longevity elder, carrying out gradient dilution on an intestinal faeces sample of the Guangxi Bama healthy and longevity elder by using a diluent, separating lactobacillus by using a MRS (lactic acid bacteria) culture medium, picking out bacterial colonies in different forms, microscopic examination and selection of gram-positive bacillus, and continuing to purify the strain by using a pouring plate method until a pure culture of the strain is obtained by separation. Through the separation process, a lactobacillus strain is obtained.
The preparation method of the diluent used in the dilution step comprises the following steps: 4.5g of sodium dihydrogen phosphate, 6.0g of disodium hydrogen phosphate, 0.5g of L-cysteine, 0.5g of agar, 0.5mL of Tween 80, 1000. 1000 mL of distilled water, heating for dissolving, adjusting pH to 7.4-7.6, and sterilizing at 121 ℃ for 15min for later use.
The pouring plate method comprises the following steps: selecting different forms of bacterial colonies, inoculating the bacterial colonies into a liquid MRS culture medium for culturing for 12-16 hours, continuously carrying out 10 times of gradient dilution on 1mL of bacterial liquid by using a diluent, adding 1mL of diluted bacterial liquid into a sterile plate, pouring a proper amount of MRS solid culture medium, after the culture medium is solidified, inversely placing the culture medium into a 37 ℃ incubator for culturing for 48 hours, and selecting single bacterial colonies to obtain a pure culture of the bacterial strain.
2. Identification and preservation
The lactobacillus obtained by the separation is identified as lactobacillus paracasei by 16S rRNA, named as lactobacillus paracasei LC-37 and preserved in China general microbiological culture Collection center, and the preservation address is: beijing, chaoyang, north Chen Xi Lu 1, 3, china academy of sciences microbiological institute, postal code: 100101 and the preservation number is CGMCC No.14055.
EXAMPLE 2 Lactobacillus paracasei LC-37 powder
The embodiment provides a preparation method of lactobacillus paracasei LC-37 powder, which specifically comprises the following steps:
1. expansion culture of strain
Inoculating Lactobacillus paracasei LC-37 frozen strain into 10mL MRS culture medium, and culturing at 37deg.C for 12 hr to obtain seed culture solution suitable for inoculation, and microscopic examination of strain form corresponding to the strain specific form without visible miscellaneous bacteria.
2. Fermentation
The seed culture solution obtained above was inoculated into a fermentation medium at an inoculum size of 2% to 5% (v/v) (2% v/v was selected in this example), and the fermentation medium was cultured at 33℃in this example as MRS medium, stirring speed was 200rpm, pH of the medium was maintained at pH 5.0, and fermentation was performed for 20 hours. After the fermentation, the culture solution was centrifuged using a disk centrifuge at 8000rpm for 2 hours, and the cells were collected.
3. Freeze-drying
The freeze-drying protective agent is added into the collected thalli, and comprises the following raw materials in parts by weight: trehalose 1kg, skim milk powder 3kg, maltodextrin 2kg, and sodium ascorbate 0.5kg. The mass ratio of the addition of the freeze-drying protective agent to the thalli obtained by centrifugation is 0.9:1, the thalli are placed in a freeze-drying disc, the temperature is increased to 30 ℃ at minus 50 ℃ within 40 hours, the vacuum degree is 3Pa, the water activity of the final bacterial powder is lower than 0.25, and the vacuum freeze-drying is finished. Then crushing and packaging to form lactobacillus paracasei LC-37 freeze-dried powder, and mixing maltodextrin according to the viable count of the lactobacillus paracasei LC-37 freeze-dried powder in proportion to finally obtain the viable count of the lactobacillus paracasei LC-37 powder of 5 multiplied by 10 9 CFU/g。
EXAMPLE 3 Lactobacillus paracasei LC-37 powder
The embodiment provides a preparation method of lactobacillus paracasei LC-37 powder, which specifically comprises the following steps:
1. expansion culture of strain
Inoculating Lactobacillus paracasei LC-37 frozen strain into 10mL MRS culture medium, and culturing at 37deg.C for 12 hr to obtain seed culture solution suitable for inoculation, and microscopic examination of strain form corresponding to the strain specific form without visible miscellaneous bacteria.
2. Fermentation
The seed culture solution obtained above was inoculated into a fermentation medium, in this example MRS medium, at an inoculum size of 2% to 5% (v/v) (5% v/v in this example), and the culture was carried out at 40℃with stirring at 100rpm, the pH of the medium was maintained at pH6.0, and fermentation was carried out for 14 hours. After the fermentation, the culture solution was centrifuged using a disk centrifuge at 10000rpm for 1 hour, and the cells were collected.
3. Freeze-drying
The freeze-drying protective agent is added into the collected thalli, and comprises the following raw materials in parts by weight: trehalose 3kg, skim milk powder 1kg, maltodextrin 4kg, sodium ascorbate 0.1kg, and the ratio of the added amount of the freeze-drying protective agent to the thalli obtained by centrifugation is 0.5:1, and the thalli are placed into a freeze-drying tray, the temperature of the vacuum freeze-drying strip is increased to 30 ℃ at-50 ℃ within 60 hours, the vacuum degree is 50Pa, so that the water activity of the final bacterial powder is lower than 0.25, and the vacuum freeze-drying is finished. Then crushing and packaging to form lactobacillus paracasei LC-37 freeze-dried powder, and mixing maltodextrin according to the viable count of the lactobacillus paracasei LC-37 freeze-dried powder in proportion to finally obtain the lactobacillus paracasei LC-37 powder with the viable count of 5 multiplied by 10 10 CFU/g。
EXAMPLE 4 Lactobacillus paracasei LC-37 powder
The embodiment provides a preparation method of lactobacillus paracasei LC-37 powder, which specifically comprises the following steps:
1. expansion culture of strain
Inoculating Lactobacillus paracasei LC-37 frozen strain into 10mL MRS culture medium, and culturing at 37deg.C for 12 hr to obtain seed culture solution suitable for inoculation, and microscopic examination of strain form corresponding to the strain specific form without visible miscellaneous bacteria.
2. Fermentation
The seed culture solution obtained above was inoculated into a fermentation medium, in this example MRS medium, at an inoculum size of 2% to 5% (v/v) (3% v/v in this example), and the culture was carried out at 37℃with stirring at 150rpm, pH of the medium was maintained at pH 5.8, and fermentation was carried out for 17 hours. After the fermentation, the culture solution was centrifuged with a disk centrifuge at 9000rpm for 1.5 hours, and the cells were collected.
3. Freeze-drying
The freeze-drying protective agent is added into the collected thalli, and comprises the following raw materials in parts by weight: trehalose 2kg, skim milk powder 2kg, maltodextrin 3kg and sodium ascorbate 0.3kg. The ratio of the adding amount of the freeze-drying protective agent to the thalli obtained by centrifugation is 0.2:1, the thalli are placed in a freeze-drying disc, the temperature is increased to 30 ℃ below zero within 50 hours under the vacuum freeze-drying condition, the vacuum degree is 25Pa, the water activity of the final bacterial powder is lower than 0.25, and the vacuum freeze-drying is finished. Then crushing and packaging to form lactobacillus paracasei LC-37 freeze-dried powder, and mixing maltodextrin according to the viable count of the lactobacillus paracasei LC-37 freeze-dried powder in proportion to finally obtain the viable count of the lactobacillus paracasei LC-37 powder of 5 multiplied by 10 11 CFU/g。
EXAMPLE 5 Lactobacillus beverage containing Lactobacillus paracasei LC-37
The embodiment provides a preparation method of lactobacillus beverage containing lactobacillus paracasei LC-37, which comprises the following steps:
taking skimmed milk powder (110 kg) as raw material, adding glucose (25 kg) and white sugar (35 kg), sterilizing, browning at 90deg.C for 5 hr, cooling to 33deg.C, inoculating lactobacillus paracasei LC-37 strain powder prepared in example 2, and inoculating with an inoculum size of 10 7 CFU/mL (even if the final concentration of Lactobacillus paracasei LC-37 in the raw material liquid is 10) 7 CFU/mL), the fermentation temperature is 33 ℃, the fermentation is finished after long-time fermentation for 72 hours, and then the obtained fermented milk is taken as a main raw material, and a proper amount of auxiliary materials such as sugar solution, essence and the like are added, so that the viable count content of the lactobacillus paracasei LC-37-containing lactobacillus paracasei beverage is finally prepared: 5X 10 7 CFU/ml。
EXAMPLE 6 Lactobacillus beverage containing Lactobacillus paracasei LC-37
The embodiment provides a preparation method of lactobacillus beverage containing lactobacillus paracasei LC-37, which comprises the following steps:
taking skimmed milk powder (140 kg) as raw material, adding glucose (15 kg) and white sugar (50 kg), sterilizing, browning at 97deg.C for 2 hr,subsequently cooled to 40 ℃, inoculated with the lactobacillus paracasei LC-37 powder prepared in example 3 in an inoculum size of 10 4 CFU/mL (even if the final concentration of Lactobacillus paracasei LC-37 in the raw material liquid is 10) 4 CFU/mL), the fermentation temperature is 40 ℃, the fermentation is finished after long-time fermentation for 100 hours, and then the obtained fermented milk is taken as a main raw material, and a proper amount of auxiliary materials such as sugar solution, essence and the like are added, so that the viable count content of the lactobacillus paracasei LC-37-containing lactobacillus paracasei beverage is finally prepared: 1X 10 8 CFU/ml。
EXAMPLE 7 Lactobacillus beverage containing Lactobacillus paracasei LC-37
The embodiment provides a preparation method of lactobacillus beverage containing lactobacillus paracasei LC-37, which comprises the following steps:
taking skimmed milk powder (125 kg) as raw material, adding glucose (20 kg) and white sugar (42 kg), sterilizing and browning at 90-97deg.C for 2-5 hr, cooling to 33-40deg.C, inoculating lactobacillus paracasei LC-37 strain powder prepared in example 4, and inoculating with an inoculum size of 10 5 CFU/mL (i.e., final concentration of Lactobacillus paracasei LC-37 in the raw material liquid was 10) 5 CFU/mL), the fermentation temperature is 37 ℃, the fermentation is finished after long-time fermentation for 86 hours, and then the obtained fermented milk is taken as a main raw material, and a proper amount of auxiliary materials such as sugar solution, essence and the like are added, so that the viable count content in the lactobacillus paracasei LC-37-containing lactobacillus beverage is finally prepared: 5X 10 8 CFU/ml。
Experimental example
1. Subject
1.1 inclusion criteria
Selecting voluntary subjects with functional dyspepsia, long-term gastrointestinal discomfort, inappetence, early satiety, excessive qi, flatulence, emesis, chronic diarrhea due to unknown reasons, constipation, etc.
1.2 exclusionary criteria
(1) Acute diarrhea.
(2) Dyspepsia caused by severe organic lesions.
(3) Weak constitution is not acceptable for the test person.
(4) Patients with serious systemic diseases such as cardiovascular, liver, kidney and hematopoietic system.
(5) The test sample is not taken as required, and the drinking result cannot be judged.
(6) Lactose intolerance or milk allergy.
1.3 design of experiments
The experiment is screened, and the total group population is 120, and is divided into 4 groups of 30 people. Wherein, the male is 55 and the female is 65; the ages were 20-60 years, the average age was 47.8 years, and the average BMI index was 24.4.BMI index: body Mass Index (BMI).
1.4 edible dosage and time
Placing the sample in a refrigerator at 2-10deg.C, taking after lunch or supper every day, taking lactobacillus paracasei LC-37 powder in A, C groups every day, and taking one bottle of lactobacillus beverage containing lactobacillus paracasei LC-37 in B, D groups every day. The preparation is administered for 28 days. The original eating habit is not changed during the test period, and the diet is normal.
Group A: lactobacillus paracasei LC-37 powder 1 (viable count content: 5×10 prepared in example 2) 9 CFU/g, daily intake: 2g) The method comprises the steps of carrying out a first treatment on the surface of the
Group B: lactobacillus paracasei LC-37-containing lactic acid bacterium beverage 1 (viable cell count content: 5X 10 prepared in example 5) 7 CFU/ml, daily intake: 200 ml)
Group C: lactobacillus paracasei LC-37 powder 2 (viable count content: 5×10 prepared in example 3) 10 CFU/g, daily intake: 2g) The method comprises the steps of carrying out a first treatment on the surface of the
Group D: lactobacillus paracasei LC-37-containing lactic acid bacterium beverage 2 (viable cell count content: 5X 10 prepared in example 7) 8 CFU/ml, daily intake: 200 ml)
1.5 Experimental procedure
The evacuation period lasted for a total of 7 days. During this time, the volunteers were unable to drink any fermented product (including yogurt, cheese, active lactobacillus beverages, etc.), but could drink milk. Day 7 is the first stool sample and blood collection time, and clinical symptom scores were made and intestinal health surveys were filled in. The drinking period lasts for 28 days, feces are collected for the second time on the 14 th day, clinical symptom scores are made, and a health questionnaire is filled. Stool samples and blood were collected on day 28, clinical symptom scores were made, and intestinal health questionnaires were filled in.
1.6 observations index
1.6.1 safety index
And detecting blood routine and blood biochemical indexes of blood samples collected before and after drinking. The blood sample of each intervention group comprises white blood cell count, hemoglobin, average red blood cell volume, average red blood cell hemoglobin concentration, platelet count, platelet compaction, lymphocyte absolute value, neutrophil absolute value, basophil absolute value, monocyte percentage, eosinophil percentage, red blood cell count, hematocrit, average red blood cell hemoglobin amount, red blood cell volume distribution width, monocyte absolute value, eosinophil absolute value, lymphocyte percentage, neutrophil percentage, basophil percentage, urea, uric acid, glutamic-oxaloacetic transaminase, total protein, white ball ratio, triglyceride, creatinine, glutamic-pyruvic transaminase, albumin, globulin, total cholesterol, blood glucose and other blood routine and blood biochemical indexes, so that lactobacillus paracasei LC-37 and lactobacillus beverage containing lactobacillus paracasei LC-37 have no adverse effect on human body. The details are shown in tables 1 to 4 below:
table 1A blood of each group was normal and biochemical before drinking
Table 2B blood of each group before drinking was normal and biochemical
Table 3C blood of each group was normal and biochemical before drinking
Table 4D blood of each group was normal and biochemical before drinking
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1.6.2 efficacy observations
Clinical symptoms of dyspepsia before and after the test of the subjects were accurately recorded, quantitative scores were given as shown in table 5, and changes in the integral of the dyspepsia symptoms before and after the test were compared.
TABLE 5 integration of clinical symptoms
The symptom scores of abdominal pain, belch, pantothenic acid, abdominal distension, appetite, diarrhea or constipation are added to obtain the integral of the clinical symptoms of dyspepsia. Statistical analysis was performed on the clinical symptom scores of the population, and the results are shown in fig. 1-7. Figures 1-6 show the change in scores for individual dyspepsia symptoms and figure 7 shows the change in scores for total clinical symptoms of dyspepsia. As can be seen from fig. 1 to fig. 7, on day 0, the scores of the various dyspepsia symptoms such as abdominal pain, eructation and pantothenic acid and the total integral of the dyspepsia symptoms are similar, and no significant difference exists, which indicates that the groups of people have comparability. At 14 days of the intervention period, a significant decrease in dyspepsia score occurred for each group. Moreover, compared with the low-dose lactobacillus paracasei, the high-dose lactobacillus paracasei has better effect than the low-dose lactobacillus paracasei, and the lactobacillus beverage containing the same-dose lactobacillus paracasei is better than the lactobacillus paracasei powder. On day 28 of the intervention period, the dyspepsia symptom score was further reduced for each group, wherein the anorexia symptoms disappeared for all groups, and the abdominal pain, belch, pantothenic acid symptoms disappeared for group D. The total symptom integral of dyspepsia in each group is reduced from the initial 4.18-4.71 to 0.17-0.71, and the total symptom integral of dyspepsia in each group is improved remarkably.
1.6.3 detection of short chain fatty acid content in feces
Establishment of a short-chain fatty acid standard curve: ether solutions containing 5mmol/L acetic acid, 5mmol/L propionic acid, 5mmol/L butyric acid and 5mmol/L heptanoic acid were prepared by a volumetric flask, and standard solutions containing 2.5,1.25,0.625,0.312 and 0.156mmol/L acetic acid, propionic acid, butyric acid and heptanoic acid were obtained by a double dilution method. And respectively transferring 1 mu L of standard solution for gas chromatographic analysis, repeatedly measuring each concentration for 3 times to obtain the peak area of each concentration and the corresponding concentration, thereby manufacturing a standard curve, and solving a regression equation and a correlation coefficient.
Extraction of short chain fatty acid in feces: a1.5 mL screw collection tube was charged with an appropriate amount of glass beads (2-3 mm), 1mL of an ether solution containing heptanoic acid (1 mmol/L concentration) and 50. Mu.L of a hydrochloric acid solution (1 mmol/L concentration) were added, and the mixture was weighed. A sample of the subject's feces (about 0.1-0.2 g) was carefully added to the collection tube with forceps, weighed again, and the weight of the feces added was recorded. Homogenizing with homogenizer for 1min, and centrifuging for 2min at 10000 r/min. Transfer supernatant into a sample bottle with an inner cannula. Because diethyl ether has strong volatility, the operation needs to be carried out rapidly under ice bath or low temperature conditions.
Chromatographic conditions for detecting short chain fatty acid concentration in fecal samples: the detector is an FID hydrogen flame ion detector; chromatographic column DB-FFAP chromatographic column; heating program: maintaining at 50deg.C for 1min, heating to 140deg.C at 10deg.C/min, maintaining for 1min, heating to 240deg.C at 30deg.C/min, and maintaining for 2min; carrier gas (N) 2 ) The pressure is 260kPa; the flow rate of nitrogen is 40mL/min, the flow rate of hydrogen is 40mL/min, and the flow rate of air is 400mL/min; the sample injection amount is 2 mu L; the temperature was measured at 230 ℃. The N2000 chromatographic workstation collects the signals and detects them.
Detecting according to the gas chromatography conditions, calculating peak area of corresponding short chain fatty acid, making standard curve (as shown in figure 12) by external standard method, correcting with internal standard heptanoic acid concentration, calculating concentration of acetic acid, propionic acid and butyric acid in the detection sample, and converting the concentration in the fecal sample.
The test results are shown in figures 8-11, and the contents of acetic acid, propionic acid, butyric acid and total short-chain fatty acid in the human waste are not changed remarkably after the lactobacillus paracasei LC-37 powder and the lactobacillus paracasei LC-37-containing lactobacillus paracasei beverage are eaten for 14 days. After 28 days of consumption, various groups of acetic acid, propionic acid, butyric acid and total short chain fatty acids were raised to different extents. Indicating that the edible lactobacillus paracasei LC-37 fungus powder and the lactobacillus beverage containing the lactobacillus paracasei LC-37 can increase the content of short chain fatty acid in the excrement.
It is apparent that the above examples are given by way of illustration only and are not limiting of the embodiments. Other variations or modifications of the above teachings will be apparent to those of ordinary skill in the art. It is not necessary here nor is it exhaustive of all embodiments. While still being apparent from variations or modifications that may be made by those skilled in the art are within the scope of the invention.
Claims (12)
1. Lactobacillus paracasei @Lactobacillus paracasei) Use of LC-37 in the manufacture of a product for improving abdominal pain, belch, pantothenic acid, abdominal distension, inappetence or diarrhea, said lactobacillus paracasei LC-37 having been deposited in the China general microbiological culture Collection center with a deposit number of CGMCC No.14055.
2. Lactobacillus paracasei @Lactobacillus paracasei) The application of LC-37 in the product for increasing the content of short chain fatty acid in intestinal tracts, wherein the lactobacillus paracasei LC-37 is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.14055.
3. Use according to claim 2, characterized in that the short chain fatty acid is at least one of acetic acid, propionic acid and butyric acid.
4. Use according to any one of claims 1-3, wherein the product is a food or a pharmaceutical.
5. The use according to claim 4, wherein the food products include, but are not limited to, solid beverages, soy products, fruit juices, dairy products, ice cream, candy and biscuits.
6. The use according to claim 4, wherein the food product is a food product comprising lactobacillus paracasei LC-37 powder.
7. The use according to claim 6, wherein the food product is a lactic acid bacteria drink; the viable count of the Lactobacillus paracasei LC-37 in the lactobacillus beverage is more than or equal to 5 multiplied by 10 7 CFU/mL。
8. The use according to any one of claims 4-7, wherein the food product further comprises raw auxiliary materials, wherein the auxiliary materials comprise but are not limited to additives.
9. The use according to any one of claims 4 to 7, wherein the auxiliary materials in the food product include, but are not limited to, nutritional supplements.
10. The use according to any one of claims 4-7, wherein the adjuvants in the food product include, but are not limited to, at least one of sugar liquor, flavor, stabilizer, thickener, preservative, mineral, vitamin, maltodextrin, whey powder, fructooligosaccharides and resistant starch.
11. The use according to claim 4, wherein the pharmaceutical dosage form includes, but is not limited to, powder, tablet, granule, capsule, solution, emulsion or suspension.
12. The use according to claim 11, wherein the medicament further comprises a pharmaceutically acceptable adjuvant.
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