CN110973247A - Lactobacillus casei Haidilao-1-containing lactobacillus beverage and preparation method thereof - Google Patents
Lactobacillus casei Haidilao-1-containing lactobacillus beverage and preparation method thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/13—Fermented milk preparations; Treatment using microorganisms or enzymes using additives
- A23C9/1307—Milk products or derivatives; Fruit or vegetable juices; Sugars, sugar alcohols, sweeteners; Oligosaccharides; Organic acids or salts thereof or acidifying agents; Flavours, dyes or pigments; Inert or aerosol gases; Carbonation methods
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
- A23C9/1234—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt characterised by using a Lactobacillus sp. other than Lactobacillus Bulgaricus, including Bificlobacterium sp.
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/125—Casei
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- Coloring Foods And Improving Nutritive Qualities (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention provides a lactobacillus beverage containing lactobacillus casei Haidilao-1 and a preparation method thereof, wherein the lactobacillus casei Haidilao-1 is preserved in the China general microbiological culture collection management center in 2018, 5 and 7 months, and the preservation number is CGMCC NO. 15735; and the bacteria content of the lactobacillus casei Haidilao-1 is 1-10 hundred million CFU/ml. The invention evaluates the digestion promoting effect of lactobacillus beverage containing lactobacillus casei Haidilao-1, and experiments recruit people with dyspepsia to test for the beverage containing the HAIDILAO-1 lactobacillus, and compared with the beverage before the test for eating, the scores of abdominal pain, eructation, pantothenic acid, abdominal distension and appetite symptoms are remarkably reduced (p is less than 0.001), diarrhea or constipation and total scores are remarkably reduced (p is less than 0.01), the contents of acetic acid, propionic acid, butyric acid and total acid in excrement are remarkably increased (p is less than 0.01), the relative abundance of beneficial bacteria in the excrement is increased, and the relative abundance of harmful bacteria is reduced. The results show that the lactobacillus beverage containing HAIDILAO-1 has good digestion promoting effect and can regulate intestinal flora.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to a lactobacillus beverage containing lactobacillus casei Haidilao-1 and a preparation method thereof.
Background
Functional dyspepsia, or dyspepsia, is a functional gastrointestinal disorder that eliminates organic diseases of the gastrointestinal system, accompanied by symptoms such as epigastric distending pain, belching, inappetence, nausea, vomiting, and the like. The prevalence rate of dyspepsia is increased year by year, and the prevalence rate of dyspepsia accounts for 18 to 45 percent in China and approximately 20 to 50 percent in an outpatient clinic of a digestive department. Has the characteristics of recurrent bouts, difficult treatment and the like, and no better treatment method exists at present.
With the continuous and deep research on the pathogenesis of the disease in recent years, the intestinal flora imbalance and the gastrointestinal hormone secretion imbalance are proved to play key roles in the whole pathophysiological process of the disease. A large number of microorganisms exist in the intestinal tract of a human body, including conditional pathogenic bacteria (enterococcus, enterobacteria and saccharomycetes) and beneficial bacteria (lactobacillus and bifidobacterium), and under normal conditions, the two groups of bacteria maintain dynamic balance in quantity and proportion, so that an intestinal microorganism ecological balance system is formed, and the normal function of the intestinal tract is maintained. When the environment in the intestinal tract is changed and the intestinal flora is disordered, diseases of the digestive system, diseases of the endocrine system, diseases of the central nervous system, metabolic diseases and the like can be caused. Therefore, the dyspepsia symptom can be relieved by eating the probiotics, and a large amount of live bacteria can be released after the probiotics enter the intestinal tract, so that the intestinal tract mucosa is facilitated to form a flora biological barrier, the flora proportion is adjusted, and the digestive absorption function of the gastrointestinal tract can be promoted.
Lactobacillus casei Haidilao-1(Lactobacillus casei) is separated from intestinal tracts of healthy people, and has obvious probiotic function. Researches show that the HAIDILAO-1 has the functions of improving immunity, relieving allergic symptoms, relieving constipation, regulating intestinal environment of healthy people and the like, and meanwhile, the HAIDILAO-1 has excellent fermentation performance, high safety and wide application and development prospects.
Disclosure of Invention
The invention aims to add lactobacillus casei Haidilao-1 into a lactobacillus beverage to obtain the lactobacillus beverage with the functions of promoting digestion and regulating intestinal flora, thereby being effective.
The purpose of the invention is realized by the following technical scheme:
a lactobacillus beverage containing lactobacillus casei Haidilao-1 has the effects of promoting digestion and regulating intestinal flora, wherein the lactobacillus casei Haidilao-1 is preserved in the China general microbiological culture Collection center in 2018, 5 and 7 months with the preservation number of CGMCC NO. 15735; and the bacteria content of the lactobacillus casei Haidilao-1 is 1-10 hundred million CFU/ml.
Further, the lactobacillus casei Haidilao-1 is live bacteria, dead bacteria or derivatives of the strain.
Another aspect of the invention:
a preparation method of lactobacillus beverage containing lactobacillus casei Haidilao-1 comprises the following steps:
(1) material melting: adding the skim milk powder and the sugar into water, stirring and dissolving to obtain a mixed material, and keeping the temperature at 40-60 ℃;
(2) browning: heating the mixed material to 90-100 ℃, and keeping the temperature for 3-6 h;
(3) cooling and inoculating: cooling to 30-40 ℃, inoculating the lactobacillus casei Haidilao-1 at 200-;
(4) fermentation: fermenting in a fermentation tank for 60-80 h;
(5) cooling and homogenizing: cooling to 20-30 deg.C, homogenizing under 10-30 Mpa;
(6) aseptically filling into a refrigeration house.
Further, the sugar comprises white granulated sugar and glucose.
Further, the weight parts of the raw materials are as follows:
compared with the prior art, the invention has the beneficial effects that:
the invention evaluates the digestion promoting effect of lactobacillus beverage containing lactobacillus casei Haidilao-1, and experiments recruit people with dyspepsia to test for the beverage containing the HAIDILAO-1 lactobacillus, and compared with the beverage before the test for eating, the scores of abdominal pain, eructation, pantothenic acid, abdominal distension and appetite symptoms are remarkably reduced (p is less than 0.001), diarrhea or constipation and total scores are remarkably reduced (p is less than 0.01), the contents of acetic acid, propionic acid, butyric acid and total acid in excrement are remarkably increased (p is less than 0.01), the relative abundance of beneficial bacteria in the excrement is increased, and the relative abundance of harmful bacteria is reduced. The results show that the lactobacillus beverage containing HAIDILAO-1 has good digestion promoting effect and can regulate intestinal flora.
And (3) biological preservation:
the Lactobacillus casei Haidilao-1(Lactobacillus casei) is preserved in the China general microbiological culture Collection center (address: Beijing Corp. facing Yang district, North West Lu No.1, institute of microbiology, China academy of sciences, postal code: 100101) in 7.5.7.2018, and the preservation number is CGMCC No. 15735.
Drawings
FIG. 1 is a graph showing the variation of exercise fatty acid content in feces of a population before and after a test meal.
Detailed Description
The embodiment provides a lactobacillus beverage containing lactobacillus casei Haidilao-1, and the preparation method comprises the following steps:
(7) material melting: adding skimmed milk powder, white sugar and glucose into water, stirring and dissolving to obtain a mixed material, and heating to 40-60 deg.C;
(8) browning: heating the mixed material to 90-100 ℃, and keeping the temperature for 3-6 h;
(9) cooling and inoculating: cooling to 30-40 ℃, inoculating the lactobacillus casei Haidilao-1 at 200-;
(10) fermentation: fermenting in a fermentation tank for 60-80 h;
(11) cooling and homogenizing: cooling to 20-30 deg.C, homogenizing under 10-30 Mpa;
(12) aseptically filling into a refrigeration house, wherein each bottle of lactobacillus beverage contains 330ml, and the lactobacillus casei Haidilao-1 contains 100-.
The weight parts of the raw materials are as follows:
experiment recruitment dyspepsia crowd tries to eat the lactobacillus beverage containing lactobacillus casei Haidilao-1.
The subject's specific requirements are as follows:
1. inclusion criteria:
(1) those 18-65 years of age;
(2) functional dyspepsia with long-term gastrointestinal discomfort;
(3) chief complaints about poor appetite, early satiety, abundant qi, flatulence and vomiting;
(4) volunteer subjects with unexplained chronic diarrhea or constipation.
2. Exclusitor criteria:
(1) acute diarrhea;
(2) dyspepsia due to severe organic lesions;
(3) those who are weak and unable to receive the test;
(4) patients with serious systemic diseases such as cardiovascular diseases, liver diseases, kidney diseases, hemopoietic diseases, etc.;
(5) lactose intolerance or milk allergy;
(6) those taking antibiotic medicines for a long time or within 15 days before the start of the experiment;
(7) those taking probiotic bacteria or fermented products within 7 days (i.e. during the emptying period) before the start of the experiment.
And (3) experimental design:
according to the experimental plan, 32 persons are recruited, the subjects are recruited according to dyspepsia symptoms (abdominal pain, eructation, pantothenic acid, abdominal distension, appetite, diarrhea, constipation and the like), and the main factors influencing the results, such as age, gender, daily diet and the like, are considered as much as possible, so that the number of persons in each age group is kept as balanced as possible to ensure comparability in the group.
The sample was left at 4 ℃ and taken within one hour after breakfast and lunch 1 time a day, 1 bottle of lactobacillus beverage (330ml) containing lactobacillus casei Haidilao-1 as described in this example. The administration is continued for 28 days. The original diet habit was not changed during the test period, and the diet was normal.
The subjects who completed the experiment were 32 people in total. The subjects are generally presented in Table 1 before feeding.
TABLE 1 general Pre-feeding Condition of subjects
Observation indexes, experimental methods and result analysis:
1. the safety index is as follows: blood biochemical and blood routine indexes, etc.
Before and after the test eating, the indexes of blood routine and blood biochemical change are in normal range. Adverse reactions and allergic reaction symptoms of the subjects are not seen after the test feeding.
The method comprises the steps of investigating main food intakes of a subject before a test meal, in the middle stage (13 th to 14 th days) and in the end stage (27 th to 28 th days) of the test meal by adopting a diet recording method, calculating and counting average daily intakes of staple food, fruit and vegetable, meat and egg food and dietary fibers of the subject, and displaying that the average intakes of various foods and dietary fibers of the foods in the test meal are not obviously different.
2. And (3) observing the efficacy:
the subjects recorded daily intake of the test samples.
(1) Number of defecation per day: subjects recorded changes in the number of bowel movements per day;
(2) defecation condition: classifying the defecation difficulty degree (abdominal pain or anus burning sensation, falling sensation, discomfort, frequent defecation but difficult defecation and little defecation and other symptoms) into I-IV grades, and counting an integral value;
grade I (0 point): defecation is normal;
grade II (1 point): only has the feeling of falling down and discomfort;
grade III (2 points): the feeling of falling and discomfort is obvious, or frequent defecation but difficult defecation and less abdominal pain or burning sensation of the anus occur;
grade IV (3 points): the abdominal pain or the burning sensation of the anus often occur, which affects the defecation;
(3) stool characteristics
Dividing the stool character into I-III grades according to a Bristol stool character classification method;
grade I (0 point): like sausage or snake, smooth and soft; like sausage, but with cracks in its surface; soft lumps, with sharp edges (easy ejection);
grade II (1 point): sausage-shaped, but lumpy; loose block-like, rough-edged, sludge-like manure;
grade III (2 points): separated hard masses like fruit stones (not easily expelled).
(4) Before the trial drinking of the product and after the end of the drinking period, the doctor consults the study and fills the following clinical symptoms scoring table.
Clinical symptom scoring table
(5) Adverse reactions (nausea, flatulence, diarrhea, abdominal pain, abnormal stool, etc.) were recorded.
(6) Short chain fatty acid content in feces
Chromatographic conditions are as follows: the detector is an FID hydrogen flame ion detector; the chromatographic column was a 60-80 mesh (acid washed, silanized) stainless steel column (25 m. times.0.320 mm) coated with 10% FFAD and 1% H packing3PO4(ii) a Temperature rising procedure: maintaining at 80 deg.C for 1min, heating to 140 deg.C at 10 deg.C/min, maintaining for 5min, heating to 240 deg.C at 25 deg.C/min, and maintaining for 4 min; carrier gas (N)2) The pressure is 260 kPa; the nitrogen flow rate is 40mL/min, the hydrogen flow rate is 30mL/min, and the air flow rate is 400 mL/min; the sample volume is 1 mu L; the detection temperature was 280 ℃. The N2000 chromatography station collects the signal and detects it.
(7) Alteration of intestinal flora in stool
Determination of intestinal flora in feces the sample intestinal microbial diversity was identified and analyzed using 16S rDNA high channel sequencing (consigned to the shanghai meiji bio-medical science and technology ltd).
Abdominal pain, eructation, acid regurgitation, abdominal distension, appetite, diarrhea or constipation are some clinical indicators of dyspepsia, and are indicators reflecting the dyspepsia degree of patients. Compared with the food before the test eating, the scores of abdominal pain, eructation, pantothenic acid, abdominal distension and appetite symptoms are remarkably reduced after the test eating (p is less than 0.001), and the scores of diarrhea or constipation and total scores are remarkably reduced (p is less than 0.01). The results are shown in table 2, which indicates that the test feeding of the yogurt containing lactobacillus paracasei can significantly reduce the score of dyspepsia symptoms and significantly improve the dyspepsia symptoms.
TABLE 2 comparison of clinical symptom scores before and after a test meal
Short chain fatty acids are the main secondary metabolites produced by intestinal microorganisms fermenting carbohydrates, including acetic acid, propionic acid and butyric acid, and are also important substances for representing that intestinal flora influences human health and physiological functions. Physiological roles of SCFA: promoting the absorption of sodium hydrate; promoting colon cell proliferation and resisting neogenesis; providing an energy source substrate; increase intestinal blood flow; stimulating the production of gastrointestinal hormones; autonomic nerves play a role in mediating the nutrition of SCFA to the gut.
Figure 1 shows the variation of the content of exercise fatty acids in faeces of a population before and after a test meal. It can be seen that the content of acetic acid and total acids in feces increased very significantly (p <0.001) and the content of propionic acid and butyric acid increased very significantly (p <0.01) after 28 days of the test feeding. From the above analysis, it is clear that the trial feeding of yoghurt containing lactobacillus paracasei can significantly increase the content of short chain fatty acids in the faeces, which suggests that the increase of short chain fatty acids may be associated with a reduction of dyspepsia.
16S rDNA high-throughput sequencing is carried out on intestinal flora in human excrement before and after test feeding, and tables 3 and 4 show the change conditions of some beneficial bacteria and harmful bacteria in human excrement. We can see that eight beneficial genera of Bacillus (Faecalibacterium), Bifidobacterium (Bifidobacterium), rare Chlorella (Subdoligranum), Dorea, Romboutsia (Romboutsia), unclassified Lachnospiraceae (unclassified _ f __ Lachnospiraceae) and Lachnospiraceae _ NK4A136_ group belonging to the family Lachnospiraceae are increased in human feces after predation, and nine harmful genera of Bacteroides (Bacteroides), Fusicatibacter, Microbacterium (Diarister), Clostridium _ sensu _ stricoto _1, Cocalabacterium (Scolototerium), Lachnotriopsis, Achilles (Alisteries), Macrobacterium (Enterobacter) and Enterobacter (Mercaptera) are decreased compared with predate. From the above analysis, it can be known that the content of beneficial bacteria can be increased and the content of harmful bacteria can be reduced by eating the yoghurt containing lactobacillus paracasei.
TABLE 3 Change in beneficial bacteria after and before tasting
| Genus of genus Bacillus | Feature(s) | Change of state |
| Faecalibacterium | Beneficial bacteria, producing short chain fatty acids | Increase of |
| Bifidobacterium | Beneficial bacteria associated with diarrhea | Increase of |
| Subdoligranulum | Beneficial bacteria associated with constipation | Increase of |
| Dorea | Beneficial bacteria, producing short chain fatty acids | Increase of |
| Romboutsia | Beneficial bacteria associated with diarrhea | Increase of |
| Roseburia | Beneficial bacteria, producing short chain fatty acids | Increase of |
| unclassified_f__Lachnospiraceae | Beneficial bacteria associated with rectal cancer | Increase of |
| Lachnospiraceae_NK4A136_group | Protecting intestinal tract and benefiting bacteria | Increase of |
TABLE 4 Change in harmful bacteria after tasting compared with before tasting
| Genus of genus Bacillus | Feature(s) | Change of state |
| Bacteroides | Associated with intestinal inflammation, harmful bacteria | Reduction of |
| Fusicatenibacter | Associated with ulcerative colitis, harmful bacteria | Reduction of |
| Dialister | Constipation-related, harmful bacteria | Reduction of |
| Clostridium_sensu_stricto_1 | Harmful bacteria | Reduction of |
| Phascolarctobacterium | Constipation-related, harmful bacteria | Reduction of |
| Lachnoclostridium | Constipation-related, harmful bacteria | Reduction of |
| Alistipes | Associated with intestinal inflammation, harmful bacteria | Reduction of |
| Megamonas | Harmful bacteria | Reduction of |
| Enterobacter | Constipation-related, harmful bacteria | Reduction of |
In the experiment, 32 subjects were recruited, and the subjects were fed with yogurt containing Lactobacillus casei Haidilao-1 as required, and 28 days later, the scores of symptoms of abdominal pain, eructation, pantothenic acid, abdominal distension and appetite of the subjects were very significantly reduced (p <0.001), the scores of symptoms of diarrhea or constipation and total score were very significantly reduced (p <0.01), the contents of acetic acid, propionic acid, butyric acid and total acid in feces were very significantly increased (p <0.01), the relative abundance of beneficial bacteria in feces was increased, and the relative abundance of harmful bacteria was decreased.
In conclusion, the lactic acid bacteria drink containing haidliao-1 is effective in relieving functional dyspepsia, which may be associated with improvement of intestinal flora and increase of short-chain fatty acids.
Claims (5)
1. The lactobacillus beverage containing lactobacillus casei Haidilao-1 is characterized in that the lactobacillus beverage has the effects of promoting digestion and regulating intestinal flora, the lactobacillus casei Haidilao-1 is preserved in China general microbiological culture collection center in 2018, 5, 7 and with the preservation number of CGMCC NO. 15735; and the bacteria content of the lactobacillus casei Haidilao-1 is 1-10 hundred million CFU/ml.
2. The lactobacillus casei Haidilao-1-containing lactic acid bacteria beverage according to claim 1, wherein the lactobacillus casei Haidilao-1 is live bacteria, inactivated bacteria or derivatives thereof.
3. A preparation method of lactobacillus casei Haidilao-1-containing lactobacillus beverage is characterized by comprising the following steps:
(1) material melting: adding the skim milk powder and the sugar into water, stirring and dissolving to obtain a mixed material, and keeping the temperature at 40-60 ℃;
(2) browning: heating the mixed material to 90-100 ℃, and keeping the temperature for 3-6 h;
(3) cooling and inoculating: cooling to 30-40 ℃, inoculating the lactobacillus casei Haidilao-1 at 200-;
(4) fermentation: fermenting in a fermentation tank for 60-80 h;
(5) cooling and homogenizing: cooling to 20-30 deg.C, homogenizing under 10-30 Mpa;
(6) aseptically filling into a refrigeration house.
4. The method of claim 3, wherein the sugar comprises white granulated sugar and glucose.
5. The preparation method of claim 4, wherein the weight parts of the raw materials are as follows:
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