CN112626001A - Probiotic composition for promoting growth of butyric acid bacteria and application thereof - Google Patents
Probiotic composition for promoting growth of butyric acid bacteria and application thereof Download PDFInfo
- Publication number
- CN112626001A CN112626001A CN202011072726.8A CN202011072726A CN112626001A CN 112626001 A CN112626001 A CN 112626001A CN 202011072726 A CN202011072726 A CN 202011072726A CN 112626001 A CN112626001 A CN 112626001A
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- okra
- butyric acid
- acid bacteria
- composition
- probiotic composition
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Abstract
The invention relates to the field of microbial fermentation, in particular to application of a composition containing okra leavening and a probiotic composition. The invention provides application of a composition containing an okra ferment in preparing a probiotic composition for promoting growth of butyric acid bacteria, and the probiotic composition containing the okra ferment. The probiotic composition promotes the growth of butyric acid bacteria in the intestinal tract, and can reduce the waist circumference of a user and improve defecation.
Description
Technical Field
The present invention relates to the field of microbial fermentation, in particular to a probiotic composition for promoting the growth of butyric acid bacteria.
Background
Numerous medical studies have shown that the intestinal environment is closely related to the maintenance of the normal functions of the body and the prevention of diseases. The intestinal tract is not only a digestive organ but also holds a large number of immune cells against pathogens to invade the body therefrom and densely distributes nerve branches to collect physiological information to feed back to the central nerve, and thus, the health of the intestinal tract has a great influence on the human health. In recent decades, intestinal bacteria are known to reside in the mucosa of the intestine, where thousands of intestinal bacteria are found to communicate with organs of the body and give appropriate responses, such as supplying nutrients to the body, regulating the development of intestinal cells, and inducing the development of the immune system, so that intestinal bacteria are known as an essential organ of the body.
The constituent bacteria of the intestinal bacteria have specific functions, and when the composition is out of balance due to bad living habits, diseases, antibiotic use and the like, the body may lose health. For example, if an individual ingests a diet with a high fat content on a daily basis, gram-negative bacteria that are fat-loving in the large intestine proliferate in large quantities and produce Lipopolysaccharides (LPS), which cause chronic inflammation in various parts of the body through blood circulation, and may eventually cause chronic diseases such as type ii diabetes, cardiovascular diseases, and even neurodegenerative diseases and cancer. Studies have also shown that mice with disrupted gut flora have memory loss under stress. Therefore, maintaining the composition of the intestinal bacteria constant helps to maintain physical and mental health.
Intestinal bacteria which are known to be beneficial to human health are potential probiotics, such as Bifidobacterium (Bifidobacterium) and Lactobacillus (Lactobacillus) bacteria. The bacteria account for 1 to 4 percent of adult intestinal bacteria, are anaerobic gram-positive bacteria growing in large intestine, and can produce butyric acid which can help defecation and promote the growth of lactic acid bacteria. In a mouse animal experiment, when the mouse takes the butyric acid bacteria under high-fat diet, the amplitude of the weight increase of the mouse is reduced.
In view of the importance of butyric acid bacteria to maintain intestinal health, there is a need to develop a probiotic composition that promotes the growth of butyric acid bacteria in the intestine.
Disclosure of Invention
Accordingly, it is an object of the present invention to provide the use of a composition comprising okra fermentate for the preparation of a probiotic composition promoting the growth of butyric acid bacteria.
In an embodiment of the present invention, the okra fermentation product is obtained by fermenting an okra pulp and water in an air culture medium with yeast (Saccharomyces cerevisiae) and Lactobacillus (Lactobacillus plantarum) simultaneously.
In an embodiment of the present invention, the air-culture solution is prepared by mixing okra pulp and water in a ratio of 1: 1 to 1: 20, the inoculation amounts of the yeast and the lactobacillus are respectively 0.1 to 0.4 percent (w/v) and 0.2 to 0.5 percent (w/v).
In a preferred embodiment of the present invention, the composition comprising okra fermentation product further comprises an oligosaccharide selected from xylooligosaccharide, fructooligosaccharide, and a combination thereof.
In some embodiments, the composition comprising okra fermentate further comprises a sugar alcohol.
In some embodiments, the oligosaccharide is a xylooligosaccharide and the sugar alcohol is a lactitol.
In some embodiments, the weight ratio of the okra ferment, the xylooligosaccharide, and the lactitol is in the range of 3-5: 2-4: 2 to 4. In a preferred embodiment, the weight ratio of the okra ferment, the xylooligosaccharide and the lactitol is in the range of 4: 3: 3.
it is still another object of the present invention to provide a probiotic composition for promoting the growth of butyric acid bacteria, comprising an okra ferment, an oligosaccharide, and a sugar alcohol, wherein the oligosaccharide is selected from xylooligosaccharide, fructooligosaccharide, and a combination thereof.
It is still another object of the present invention to provide a use of the aforementioned prebiotic composition for the preparation of a pharmaceutical composition for reducing waist circumference.
In one embodiment of the invention, the prebiotic composition increases the number of butyric acid bacteria in the gut of an obese subject and decreases the weight, body fat mass and waist circumference of an obese subject.
Disclosed herein are various prebiotics and combinations thereof that are beneficial to the growth of butyric acid bacteria. The probiotic combination may be used to prepare a probiotic composition effective in promoting the growth of butyric acid bacteria, for example as a culture medium supplement for in vitro bacterial culture, or as a probiotic product promoting the proliferation of butyric acid bacteria in the intestinal tract of a living body, such as a human. The prebiotic composition may be in the form of a powder, granule, solution, or capsule, and may be formulated as a food, beverage, pharmaceutical, or nutritional supplement for oral administration to an individual.
The following embodiments are provided to illustrate the features and applications of the present invention, rather than to limit the scope of the invention, and the invention is not limited thereto.
The invention is described in detail below with reference to the drawings and specific examples, but the invention is not limited thereto.
Drawings
FIG. 1 shows the relative counts of butyric acid bacteria grown anaerobically for 24 hours in medium with or without prebiotics.
Fig. 2A shows the statistics of the average number of bowel movements before the subjects took the probiotic composition according to one embodiment of the present invention.
Fig. 2B shows the statistics of the average number of days to defecate after a subject took one embodiment of the probiotic composition for four weeks.
Fig. 3A shows the statistical result of how often a subject experiences unclean defecation before taking a prebiotic composition according to one embodiment of the invention.
Fig. 3B shows statistics of how often a subject experiences unclean bowel movements after taking a probiotic composition according to one embodiment of the invention for four weeks.
Fig. 4A shows statistical results of a stool condition of a subject prior to administration of a prebiotic composition according to one embodiment of the invention.
Fig. 4B shows statistical results of a stool condition after a subject takes a prebiotic composition according to one embodiment of the invention for four weeks.
Fig. 5A shows the statistical results of the voluntary defecation discharge before the subjects took the probiotic composition according to one embodiment of the invention.
Fig. 5B shows the statistical results of the voluntary defecation discharge after the subjects took one week with the prebiotic composition of one embodiment of the present invention.
Fig. 6 shows the mean waist circumference variation of a subject after four weeks of administration of a prebiotic composition according to one embodiment of the invention.
Fig. 7 shows the mean body weight change of a subject after four weeks of administration of a prebiotic composition according to one embodiment of the invention.
Fig. 8 shows the change in body fat rate of a subject after four weeks of administration of a prebiotic composition according to one embodiment of the invention.
Detailed Description
The invention provides a use of a composition comprising okra fermentate for the preparation of a probiotic composition for promoting the growth of butyric acid bacteria. In addition to the okra ferment, the probiotic composition may further comprise an oligosaccharide selected from the group consisting of xylooligosaccharides, fructooligosaccharides, and combinations thereof, and a sugar alcohol, such as sorbitol, mannitol, erythritol, maltitol, lactitol, and xylitol. The prebiotic composition preferably comprises okra fermentation product, xylooligosaccharide and lactitol, and the weight ratio of the okra fermentation product to the xylooligosaccharide is preferably 3-5: 2-4: 2 to 4. The following examples show the growth promoting effect of each component and combinations thereof in the prebiotic composition on butyric acid bacteria, and also show that the prebiotic composition can reduce body weight, body fat percentage, and waist circumference of obese subjects.
Definition of
As used herein, the numerical values are approximations and all numerical data are reported to be within the 20 percent range, preferably within the 10 percent range, and most preferably within the 5 percent range.
The term "okra" as used herein refers to a one-or perennial herb of the genus okra (Abelmoschus) of the family Malvaceae (Malvaceae), also known as Abelmoschus manihot, carob bean, solanum lyratum, anantia malva, cabochloa, okra, vietnamese sesame, and cavel.
As used herein, "xylo-oligosaccharide" refers to an oligosaccharide comprising 3 to 10 xylose (xylose) units, which may be derived from plants, or may be obtained by microbial fermentation, enzymatic conversion, or chemical synthesis.
According to the present invention, the food product may be used as a food additive (food additive) to be added during the preparation of the raw material or during the preparation of the food by conventional methods, and formulated with any edible material into a food product for ingestion by humans and non-human animals.
According to the present invention, the types of food products include, but are not limited to: beverages (leafages), fermented foods (fermented foods), bakery products (bakery products), health foods (health foods) and dietary supplements (dietary supplements).
As used herein, the terms "yeast (Saccharomyces cerevisiae)" and "lactic acid bacteria (Lactobacillus plantarum)" are intended to encompass brewer's yeast and Lactobacillus plantarum, respectively, which are readily available to those skilled in the art, e.g., from domestic or foreign depositories, or brewer's yeast and Lactobacillus plantarum strains that have been isolated and purified from natural sources using microbial isolation procedures customary in the art.
Materials and methods
Preparation of okra fermentation product
Okra (Abelmoschus esculentus) is a perennial herb of the genus Abelmoschus (Abelmoschus) of the family Malvaceae (Malvaceae), and is also called Abelmoschus manihot, carob bean, solanum lyratum, anapula, podocarpus shikoshikohlii, okra, vietnamese sesame, and cavel bean. Originally produced in Abies, Egypt and Caribbean in northeast Africa or tropical Asia, and introduced into Taiwan in 1901, the producing areas are distributed in Zhanghua, Yunling and Jiayi counties, and the full producing period is four to September every year. The okra is erect, about one meter higher, the whole plant is villous, the stem is lignified, the flower is complete and is faint yellow, tender fruits can be harvested three to four days after blossom withering, the okra fruits are capsules, about ten centimeters long, slender and similar to goat horn or hot pepper, the peel is thin and leather, the tip is sharp, and the color of the fruits is from light green to dark green and also has purplish red. The okra fruits contain special sticky and slippery substances and have special fragrance or flavor. The invention ferments okra to obtain a leavening with more abundant composition molecules.
The okra fermentations used in examples 1-3 herein were prepared as follows. Firstly, okra pulp and water are mixed according to the weight ratio of 1: 1 to 1: 20, preferably 1: 1 to 1: 5 mixing to obtain an air culture solution. And (3) after the air culture solution is treated by a pasteurization method, cooling to room temperature for subsequent fermentation. Then, 0.01-0.5% (w/w) of yeast (Saccharomyces cerevisiae, purchased from center for biological resources preservation and research, Taiwan, No. BCRC20271) and 0.01-0.25% (w/w) of lactic acid bacteria (Lactobacillus plantarum, purchased from center for biological resources preservation and research, Taiwan, No. BCRC910760) were simultaneously implanted into the air culture solution, and fermentation was performed at 25-35 ℃ for 1-7 days, with actual time varying depending on the fermentation state. Finally, under the condition that the two bacteria are not removed, the set specifications of the sugar degree range of 2-4 degrees, the pH value of <4, the alcohol content of 5 percent and the like are used, if the test conforms to the specifications, the completion of the fermentation is judged, and the obtained fermentation product is also an experimental sample of the okra fermentation product used in the examples 1-3. Then, the fermentation liquid is decompressed and concentrated at 45-70 ℃, filtered by a sieve with 200-400mesh, added with 40-70% isomalto-oligosaccharide to adjust the specification, and heated at 95-105 ℃ for 70-90 minutes for sterilization. The okra fermentation liquid can be prepared into a powdery product by a drying method such as spray drying.
Oligosaccharide, polysaccharide and sugar alcohol
Oligosaccharides used in the examples herein include xylooligosaccharides (available from Shandong Longli Biotech, Inc.), fructooligosaccharides (available from MEIJI CO., Ltd.). The polysaccharide used included inulin (available from Cosucra group Horconing SA). The sugar alcohols used include lactitol (available from Hongwei Biotech, Inc.), sorbitol, mannitol, erythritol, maltitol, and xylitol.
Butyric acid bacteria source and sample preparation
The following examples use butyric acid bacteria (BCRC 81048) purchased from the biological resources conservation and Research Center (BCRC) of the institute of food industry development.
An activation procedure is performed to activate the butyric acid bacteria. First, a butyric acid bacterium was cultured in a 10 vol% inoculum size in a liquid lactobacillus MRS medium (BD Difco)TMLactobacillus MRS Broth, product No. DF0881-17-5, from Thermo Fischer Scientific)) and cultured for 16 hours at 37 ℃ to activate butyric acid bacteria. After thawing and activation, the bacteria can be used for probiotic testing.
Example 1
Probiotic testing of butyric acid bacteria
To identify nutrients beneficial to the growth of butyric acid bacteria, pre-cultured butyric acid bacteria were inoculated with 3% inoculum size into MRS medium containing 1% (w/w) candidate prebiotics, cultured at 37 ℃ under anaerobic conditions for 24 hours, and the resulting bacterial fluid was spread on MRS medium and cultured for 72 hours to estimate the number of bacteria. The candidate prebiotics are xylo-oligosaccharides, lactitol, inulin, fructo-oligosaccharides, or okra fermentate. As a control, butyric acid bacteria were additionally cultured in MRS medium without candidate prebiotics (blank).
FIG. 1 shows the effect of various candidate probiotics on the growth of butyric acid bacteria, and the turbidity of MRS culture medium is measured 72 hours later by using a spectrophotometer, wherein the wavelength is selected to be 600nm to measure the viable bacteria concentration of the butyric acid bacteria. According to FIG. 1, the blank set had an OD600 of 1.1. Compared with the blank group, the candidate probiotics with obviously improved bacterial count comprise okra oligosaccharide, inulin, fructooligosaccharide and okra fermentation products, and the OD600 is respectively improved to about 2.2, 1.8, 1.7 and 2.9. This result demonstrates that any of okra fermentate, xylo-oligosaccharide, fructo-oligosaccharide, and lactitol can be used as the butyrate prebiotic.
Example 2
Probiotic composition for promoting growth of butyric acid bacteria
To evaluate the growth promoting effect of different combinations of the probiotics of the butyric acid bacteria on the growth of the butyric acid bacteria, the pre-cultured butyric acid bacteria are added into an MRS culture medium containing 1% (w/w) of the probiotic composition according to the inoculation amount of 3%, the mixture is cultured for 24 hours under the anaerobic condition at 37 ℃, and then the obtained bacterial liquid is coated on the MRS agar culture medium and cultured for 72 hours to estimate the bacterial count. The probiotic composition in MRS medium had the following composition: 0.4% okra ferment, 0.3% xylo-oligosaccharide, and 0.3% lactitol. As a control, butyric acid bacteria were additionally cultured in MRS medium without prebiotics (blank).
Figure 1 shows the effect of the aforementioned probiotic composition on the growth of butyric acid bacteria, the number of bacteria OD600 of the blank group being 1.1. According to FIG. 1, the addition of the probiotic composition significantly increased the relative count of butyric acid bacteria to about 3.4, respectively, compared to the blank control group. Therefore, it is known that the weight ratio of the combination of okra fermentation product, xylooligosaccharide, and lactitol is 3 to 5: 2-4: 2 to 4, the growth of butyric acid bacteria can be most effectively promoted, even though the growth promoting effect of lactitol is the lowest as shown in FIG. 1.
Example 3
Physiological effects of probiotic compositions
This example discusses the physiological effects of continuous administration of a prebiotic composition capsule containing 200mg okra fermentate, 150mg xylooligosaccharide, and 150mg lactitol on subjects who were healthy males or females with a BMI greater than 24 or a body fat greater than 25. Nine subjects of 20 to 55 years old were orally administered the probiotic composition for four consecutive weeks after daily lunch at a dose of 400 mg/person/day and evaluated for difficulty in defecation including average number of days to defecate, frequency of clean defecation, state of stools and smoothness of defecation before and after administration for four weeks, and waist circumference, weight and body fat were measured on a body composition meter (TANITA BC-545F).
To confirm that the okra ferment or the composition thereof of the present invention has the effect of improving the degree of difficulty in defecation, the subjects filled out a gastrointestinal tract questionnaire (table one) and a defecation difficulty degree questionnaire (table two) before and after taking the probiotic composition for four weeks. Fig. 2A to 5B show the experimental results counted by the subjects who completed table one and table two.
TABLE-gastrointestinal tract questionnaire
TABLE II _ difficulty level questionnaire (please check in the frame) (Single selection)
Please refer to fig. 2A and fig. 2B, which are statistics of results before and after the subject took the capsule for four weeks, respectively, according to the first question in table i. Referring to fig. 2A, 22% of subjects had a bowel movement number of three days before taking the capsule. Referring to fig. 2B, after the subjects took the capsule for four weeks, the number of the subjects who had stood once every three days was reduced to 0%, the number of the subjects who had stood once every day was increased to 45%, and the number of the subjects who had stood twice every day was increased to 22%, which revealed that the probiotic composition capsule of the present invention had a significant improvement in the number of days to stools.
Please refer to fig. 3A and fig. 3B, which are statistics of results before and after the subject took the capsule for four weeks, respectively, according to the fourth item in table one. Referring to fig. 3A, 11% of subjects had trouble with always unclean defecation and 78% of subjects had trouble with unclean defecation before they took the capsules. Referring to fig. 3B, after the subject took the capsule for four weeks, the subject who always experienced unclean defecation and often experienced unclean defecation decreased to 0%, and all subjects experienced little trouble of unclean defecation. Therefore, it was found that the prebiotic composition capsules of the present invention were effective in improving the frequency of unclean defecation.
Referring to fig. 4A and 4B, statistics of the results of the subjects before and after taking the capsule for four weeks are shown according to the sixth item in table i. Referring to fig. 4A, prior to the administration of the capsule, 56% of the subjects had moderate stool conditions and 33% of the subjects had mild stool conditions. Referring to fig. 4B, after the subjects took the capsule for four weeks, subjects with moderate stool conditions decreased to 11%, and subjects with mild and no stool conditions increased to 56% and 33%, respectively. Therefore, it can be seen that the prebiotic composition capsules of the present invention can improve the condition of stubborn stool.
Referring to fig. 5A and 5B, the results of the subjects before and after taking the capsule for four weeks are counted according to the rectal symptoms of table two, respectively. Referring to fig. 5A, 56% of subjects experienced moderate difficulty when they had a bowel movement, and 11% experienced mild difficulty when they had a bowel movement, before taking the capsule. Referring to fig. 5B, after the subjects took the capsule for four weeks, only 11% of the subjects felt moderate difficulty when they had a bowel movement, and the remaining 89% of the subjects felt no difficulty when they had a bowel movement. Therefore, it can be seen that the prebiotic composition capsule of the present invention is effective in improving the degree of defecation smoothness.
Referring to fig. 6-8, administration of the probiotic composition for four weeks reduced the average waist circumference by about 1 cm, reduced the average body weight by about 0.3 kg, and reduced the average body fat percentage by about 0.2% compared to administration prior to administration. This result demonstrates that long term use of the probiotic composition of the invention can improve the body condition of obese persons.
In summary, the present invention discloses various prebiotics and combinations thereof that are beneficial to the growth of butyric acid bacteria. The probiotic combination may be used to prepare a probiotic composition for promoting the growth of butyric acid bacteria, such as a medium supplement for in vitro bacterial culture, or a probiotic product for promoting the proliferation of butyric acid bacteria in the human intestinal tract. The prebiotic composition may be in the form of a powder, granule, solution, or capsule, and may be formulated as a food, beverage, pharmaceutical, or nutritional supplement for oral administration to an individual.
The present invention is capable of other embodiments, and various changes and modifications may be made by one skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (10)
1. Use of a composition comprising an okra ferment for the manufacture of a probiotic composition for promoting the growth of butyric acid bacteria.
2. The use as claimed in claim 1, wherein the okra fermented product is obtained by fermenting okra pulp and water in an air culture medium with a yeast and a lactobacillus simultaneously.
3. Use according to claim 2, wherein the air culture medium is a mixture of pulp of Musa spp. 1 to 1: 20 by weight ratio.
4. The use according to claim 2, wherein the yeast and the lactobacillus are inoculated in amounts of 0.1% to 0.4% (w/v) and 0.2% to 0.5% (w/v), respectively.
5. The use of claim 1, wherein the composition comprising okra fermentate further comprises an oligosaccharide selected from the group consisting of xylo-oligosaccharides, fructo-oligosaccharides, and combinations thereof.
6. The use of claim 5, wherein the composition comprising okra ferment further comprises a sugar alcohol.
7. The use according to claim 6, characterized in that the oligosaccharide is a xylooligosaccharide and the sugar alcohol is a lactitol.
8. The use according to claim 7, wherein the weight ratio of the okra ferment, the xylooligosaccharide and the lactitol is in the range of 3-5: 2-4: 2 to 4.
9. A probiotic composition for promoting the growth of butyric acid bacteria, comprising an okra ferment, an oligosaccharide, and a sugar alcohol, wherein the oligosaccharide is selected from the group consisting of xylooligosaccharide, fructooligosaccharide, and combinations thereof.
10. Use of a probiotic composition according to claim 9 for the preparation of a pharmaceutical composition for reducing waist circumference.
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| CN102827720A (en) * | 2011-06-14 | 2012-12-19 | 如意情集团股份有限公司 | Preparation technology of okra wine |
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| TW202114542A (en) | 2021-04-16 |
| CN112618427B (en) | 2023-01-24 |
| TWI744028B (en) | 2021-10-21 |
| TWI757911B (en) | 2022-03-11 |
| CN112618427A (en) | 2021-04-09 |
| TW202115241A (en) | 2021-04-16 |
| CN112694985A (en) | 2021-04-23 |
| CN112694985B (en) | 2022-12-30 |
| TWI784326B (en) | 2022-11-21 |
| CN112618610A (en) | 2021-04-09 |
| TW202115244A (en) | 2021-04-16 |
| CN112626001B (en) | 2022-12-27 |
| TW202114631A (en) | 2021-04-16 |
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