CN114600986A - Hypoglycemic effect of kombucha prepared by direct-throwing starter and kombucha - Google Patents

Abstract

The invention discloses a process for preparing kombucha with a direct-throwing starter and the hypoglycemic effect of the kombucha. In the present invention, the kombucha product is fermented by a starter prepared from the dominant strains Jojuformella C82, Bayer combined yeast Y4 and Lactobacillus plantarum L10 isolated from traditional kombucha, the strain is stable and the difference in fermentation flavor is not large, which is beneficial to Industrialization of kombucha beverages. The invention provides the application of a kombucha product in medicine or food for improving type II diabetes, which can significantly improve type II diabetes and improve the hypoglycemic effect.

Description

Hypoglycemic effect of kombucha prepared by direct-throwing starter and kombucha

technical field

The invention belongs to the field of industrial production methods of tea beverages, and particularly relates to a direct-throwing kombucha starter and the application of the hypoglycemic effect of kombucha.

Background technique

Kombucha is a kind of drink that is beneficial to the human body produced by the mixed fermentation of sugar, tea and water by lactic acid bacteria, yeast and acetic acid bacteria. Because the kombucha bacterial membrane is similar to the jellyfish skin, the kombucha is called "Haibao", and because its bacterial liquid has the effect of promoting digestion, it is also called "weibao". Due to the lack of knowledge of microbiology and technology, the fermented kombucha is susceptible to contamination by miscellaneous bacteria, so it cannot be widely used in industrial production. Kombucha has a variety of nutrients, including organic acids, phenolic substances, amino acids, etc., such as glucuronic acid. It has a detoxification effect and helps protect human health and prevent chronic non-communicable diseases.

Diabetes Mellitus (DM) is a metabolic disease associated with impaired insulin secretion, insulin resistance and pancreatic β-cell dysfunction. The body has chronic inflammation, glucose and lipid metabolism disorders, and oxidative damage. And "three more and one less" as the main symbol. At present, with the rapid development of modern society, people's lifestyles, especially eating habits, have undergone tremendous changes. Excessive intake of sugar and fat in the diet can lead to an increased incidence of chronic metabolic diseases such as obesity and diabetes. The main place for the absorption of tea polyphenols, which are fermented metabolites of kombucha, is in the intestine, and tea polyphenols are closely related to the intestinal flora. Literature reports indicate that the intestinal flora can extensively metabolize polyphenols and affect their biological activities. Similarly, polyphenols such as catechins, flavonoids and phenolic acids can also regulate the intestinal flora of type II diabetes. disorder phenomenon.

In recent years, domestic scholars have conducted more and more researches on kombucha, including kombucha fermented beverages, fermented snack foods, application of bacteria in kombucha, fermentation process and fermentation equipment, etc. For example, the patent publication with the application number CN201811389231.0 A kind of Huangpi fruit and kombucha compound beverage and a preparation method thereof are disclosed; for example, the patent with the application number of CN201910546380.1 discloses a kombucha tofu and a production method thereof. There are also relevant reports on the research and development of direct-injection starter. For example, the patent with the application number of CN201811311727.6 discloses a preparation method of a direct-injection kombucha starter.

Through searching, no patent publications related to the patent application of the present invention have been found.

SUMMARY OF THE INVENTION

The purpose of the present invention is to overcome the problems of complex microbial phase caused by the mixed bacterial strain fermentation of the existing fermented tea beverage, the fermentation process is susceptible to contamination by miscellaneous bacteria, and the product flavor is not uniform, and provides a pure-bred fermented kombucha beverage process. The present invention adopts the strain as the dominant strain isolated from the traditional kombucha, which ensures that the present invention and the traditional kombucha remain similar in flavor and nutrients.

Another object of the present invention is to use Kunming mice as model animals for kombucha products with hypoglycemic effect and T2DM medicines and/or food, kombucha products can significantly improve T2DM symptoms and improve hypoglycemic effect.

The technical scheme that the present invention realizes the object is:

Hypoglycemic effect of kombucha prepared by direct-throwing starter and kombucha.

And, described Komagataeibacter xylinus (Komataeibacter xylinus) C82, Bayer combined yeast (Zygosaccharomyces bailii) Y4 are separated and obtained with traditional kombucha, have been preserved in the General Microorganism Center of the China Microorganism Culture Collection Management Committee, and the preservation number is respectively CGMCC No.16184, CGMCC No.16183. Lactobacillus plantarum BC299 is preserved in the General Microbiology Center of China Microorganism Culture Collection Management Committee, and the preservation number is CGMCC No.6326.

Moreover, the medicine or food is made into a dosage form acceptable in medicine and/or food by using Trichoderma C82, Bayer combined yeast Y4 and Lactobacillus plantarum BC 299 viable bacteria as active ingredients.

And, the direct-throwing starter prepares kombucha, comprising the following steps:

1) Selection of tea leaves: select dry green tea, black tea or other tea leaves with strong tea fragrance, and the mass percentage of tea leaves is 0.8%.

2) Extraction and filtration: put the tea leaves into boiling water, boil for 5 minutes, and filter with edible filter cloth to obtain the extract.

3) Sterilization: After adding 4-10% white granulated sugar to the filtered extract, sterilize at 115°C for 15 minutes.

4) Fermentation: The starter of Trichomoniasis C82, Bayer combined yeast Y4 and Lactobacillus plantarum BC299 was put into the sterilized tea soup at ^106-8 CFU/100mL, and fermented at 30°C for 4-7d.

5) Centrifugation: use 4°C, 8000r, 20min centrifugation to remove bacterial cells.

6) Filling: mix the kombucha stock solution with 0-5 times water after centrifugation, and fill it to obtain a kombucha beverage.

And, the described application, the purpose that its tea leaves carry out leaching is that the purpose that tea leaves is carried out leaching is that the tea polyphenols in the tea leaves and other polyphenolic components such as catechins, flavonoids and phenolic acids and other components Extract into tea soup.

Moreover, the application is characterized in that: compared with traditional kombucha, the content of short-chain fatty acids such as formic acid, acetic acid and butyric acid in the kombucha beverage is increased, the fasting blood glucose concentration of the kombucha beverage is reduced, and the blood sugar is lowered. effect increased.

Moreover, in the application, the product function is characterized in that: the important pathological features of the type II diabetes include relieving the symptoms of "three more and one less", lowering blood sugar concentration, improving glucose tolerance, and improving diabetes in mice. Weight loss phenomenon. That is, kombucha products can improve type II diabetes and improve hypoglycemic effect by increasing the body weight of mice, reducing diet, water intake, urine output, reducing fasting blood sugar and improving glucose tolerance.

Moreover, the application is characterized in that: the "three more and one less" includes increased water intake, increased dietary intake, increased urine output, and decreased body weight.

A medicine or food with hypoglycemic effect is characterized in that: the medicine or food comprises an effective amount of kombucha and auxiliary materials acceptable in medicine and food.

Moreover, the dosage forms of the medicine or food include powder, tablet, granule, capsule and drink.

The advantages and positive effects of the present invention are:

1. The present invention adopts pure-bred fermentation to improve the problems of complex microbial phase caused by fermentation of natural strains, susceptibility to contamination by miscellaneous bacteria during fermentation, and uneven product flavor.

2. The present invention utilizes the determination of "three more and one less" related indicators to measure the indicators of body weight, food intake and water intake of diabetic mice fed with kombucha beverages. The body weight of the mice in each group was tested, and the results showed that the kombucha fermented liquid could improve the weight loss of diabetic mice, and the kombucha fermented liquid was better than the tea soup. The diet and diet of mice in each group were tested, and the results showed that kombucha fermented liquid could significantly reduce the diet and water intake of diabetic mice.

3. The present invention also detects the changes of fasting blood glucose and the content of glycated serum protein in diabetic mice fed with kombucha beverages. Compared with the T2DM mice, the fasting blood sugar of the mice fed by fermentation with kombucha is significantly reduced;

Description of drawings

Figure 1 shows the experimental results of the effect of test samples on the content of glycated serum protein in diabetic mice: data are mean ± S.D; NC is normal group, DC is model group, MET is metformin group, KT is kombucha group, TS is It is the tea soup group; ##P<0.01 means there is a very significant difference between the sample group and the DC group, **P<0.01 means there is a very significant difference between the DC group and the NC group.

Detailed ways

The embodiments of the present invention will be described in detail below. It should be noted that the embodiments are descriptive, not restrictive, and cannot limit the protection scope of the present invention.

The raw materials used in the present invention, unless otherwise specified, are conventional commercial products, the methods used in the present invention, unless otherwise specified, are conventional methods in the field, and the quality of each substance used in the present invention is conventionally used quality.

Example 1

A kind of kombucha beverage process of the present invention

1 raw material

Tea, Trichoderma C82 starter, Bayer combined yeast Y4 starter, Lactobacillus plantarum BC299 starter, sugar, water.

2 Preparation methods

1) Selection of tea leaves: select dry green tea, black tea or other tea leaves with strong tea fragrance, and the mass percentage of tea leaves is 0.8%.

2) Extraction and filtration: put the tea leaves into boiling water, boil for 5 minutes, and filter with edible filter cloth to obtain the extract.

3) Sterilization: After adding 4-10% white granulated sugar to the filtered extract, sterilize at 115°C for 15 minutes.

4) Fermentation: The starter of Trichomoniasis C82, Bayer combined yeast Y4 and Lactobacillus plantarum BC299 was put into the sterilized tea soup at ^106-8 CFU/100mL, and fermented at 30°C for 4-7d.

5) Centrifugation: use 4°C, 8000r, 20min centrifugation to remove bacterial cells.

6) Filling: mix the kombucha stock solution with 0-5 times water after centrifugation, and fill it to obtain a kombucha beverage.

3 Product indicators

Table 1 Relevant indicators of kombucha beverages

This product uses pure-bred fermented kombucha in its total acid and total phenolic content, which is significantly higher than traditional kombucha, and its ability to metabolize sugar is better than traditional kombucha. Compared with traditional kombucha, the present invention has obvious improvement in short-chain fatty acid, especially acetic acid and butyric acid. Acetic acid is a sour agent, which can not only improve the taste of products, but also prevent the growth of miscellaneous bacteria. The direct intake of short-chain fatty acids can effectively improve the hypoglycemic effect of the body. Therefore, the present invention can improve the hypoglycemic effect, and the following experiments are carried out to verify.

Example 2

Configuration of tea soup and kombucha fermentation broth:

1) Tea soup (TS): the extract in Example 1 was used.

2) Preparation of kombucha fermentation broth (KT): The kombucha fermentation broth in Example 1 was used.

Example 3

Kunming mice were used as experimental materials, and the experiment was started after 1 week of adaptive feeding. T2DM model mice were induced by high-sugar and high-fat diet combined with streptozotocin (STZ). After being fed with high-sugar and high-fat diet for 4 weeks, the mice were fasted for 12 hours, and the mice were injected with STZ in small doses at 70 mg/kg body weight for 5 consecutive times (48 hours between each injection). /L mice were incorporated into the diabetes model. The mice were divided into 5 groups (10 in each group): diabetes model group, normal control group, metformin positive control group (0.13g/kg), new type of kombucha, tea soup; 5 groups were all at 11.1ml/kg, 1 The mice in the normal control group and the diabetic model group were given normal saline by gavage.

Table 2 Grouping and administration of experimental animals

Table 3 Composition of high-sugar and high-fat feed

Example 4

The body weight of mice in each group was observed before and after modeling and after administration. The results are shown in Table 4. Table 4 shows: before modeling (injection of STZ), the body weight of mice in DC group increased to 1.3 times that of NC group, and the difference was statistically significant (p<0.01). There was no significant difference in body weight between other groups and DC group. After modeling (injection of STZ), the body weight of the mice in the DC group was significantly lower than that of the mice in the NC group. With the passage of the administration time, to the 4th week of administration, the body weight of the mice in the MET group and the KT group increased by 1.24 times and 1.23 times, respectively, compared with the DC group, and the difference was statistically significant (p < 0.01), indicating that the fermentation of kombucha The kombucha fermented liquid can improve the weight loss of diabetic mice, and the effect of kombucha fermented liquid is better than that of tea soup.

Table 4 Effects of tested samples on body weight changes of T2DM mice

Note: NC is the normal group, DC is the model group, MET is the metformin group, KT is the kombucha group, and TS is the tea decoction group; ^## P＜0.0 means there is a very significant difference between the sample group and the DC group, **P＜ 0.01 means that there is a very significant difference between the DC group and the NC group.

Example 5

The changes of diet and water intake of mice in each group were observed before and after model establishment and after administration. The results are shown in Table 5 and Table 6. Table 5 and Table 6 show that: after modeling, the average daily food intake and water intake of the mice in the DC group increased significantly; with the passage of the administration time, in the fourth week of gavage, the dietary intake and water intake of the mice in each group were administered All were extremely significant or significantly lower than that of DC group mice (p<0.01 or p<0.05).

Table 5 Effects of tested samples on average daily dietary intake of T2DM mice

Note: NC is the normal group, DC is the model group, MET is the metformin group, KT is the kombucha group, and TS is the tea soup group; ^## P＜0.01 means there is a very significant difference between the sample group and the DC group, **P＜ 0.01 means that there is a very significant difference between the DC group and the NC group.

Table 6 Effects of tested samples on the average daily water intake of T2DM mice

Note: NC is the normal group, DC is the model group, MET is the metformin group, KT is the kombucha group, and TS is the tea decoction group; ^## P＜0.0 means there is a very significant difference between the sample group and the DC group, **P＜ 0.01 means that there is a very significant difference between the DC group and the NC group.

In Examples 4 and 5, the body weight, diet and water consumption were detected, and it was shown that kombucha significantly improved the symptoms of three excess and one less in diabetic mice, and the effect was similar to that of metformin, and the difference was not significant.

Example 6

Fasting blood glucose was used to observe the changes of blood glucose concentration of mice in each group before and after modeling and after administration. The results are shown in Table 7. Table 7 shows that the FBG of each group after STZ injection was 4 times higher than that of the NC group (P<0.01), and the blood glucose level of the DC group was stable between 20-22 mmol/L. Compared with the DC group, the dietary intake of the mice in the MT and TS groups was reduced by 33.2%, 29.1%, and 14.7%, respectively. It can be seen that kombucha can effectively improve the hyperglycemia characteristics of diabetic mice. Significantly.

Table 7 Comparison of fasting blood glucose of mice in each group

Note: NC is the normal group, DC is the model group, MET is the metformin group, KT is the kombucha group, and TS is the tea soup group; ^## P＜0.01 means there is a very significant difference between the sample group and the DC group, **P＜ 0.01 means that there is a very significant difference between the DC group and the NC group.

Example 7

The glycated serum protein detection was used to observe the changes of the glycated serum protein content of the mice in each group, and the GSP content of the T2DM mice was detected by the glycated serum protein kit. The results are shown in Figure 1. Figure 1 shows: Compared with the NC group, the GSP content of the DC group was increased by 2.16 times (p<0.01), which was 2.68 mmol/L; 51.2%, 45.7%, 21.4% (P<0.01), and their contents were 1.31 mmol/L, 1.69 mmol/L and 2.11 mmol/L, respectively. It can be seen that the blood glucose of T2DM mice is maintained at a relatively high level, resulting in the formation of a large amount of fructosamine. The new type of kombucha can effectively improve the glucose tolerance of diabetic mice, and the effect is the best.

Conclusion: The kombucha C82 and Bayer combined with yeast Y4 were isolated from traditional kombucha to prepare starter, and Lactobacillus plantarum BC299 was used to ferment kombucha products, and kombucha products could increase the weight of mice, reduce the amount of diet and water consumption. It can improve type II diabetes mellitus and improve hypoglycemic effect by reducing fasting blood glucose and improving glucose tolerance.

Note: The present invention uses SPSS to analyze the differences between the experimental groups. All data presented herein are presented as mean ± standard deviation (SD). ^## means there is a very significant difference between the sample group and the DC group, P<0.01, the difference is extremely significant; ^** means there is a very significant difference between the DC group and the NC group, P<0.01, the difference is extremely significant.

Although the embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that various substitutions, changes and modifications are possible without departing from the spirit and scope of the invention and the appended claims, therefore , the scope of the present invention is not limited to the contents disclosed in the embodiments.

Claims (10)

1. The direct-vat-set starter is used for preparing the black tea fungus and the blood sugar reducing function of the black tea fungus.

2. Use according to claim 1, characterized in that: the colt shaped bacteria (Komagataeibacter xylinus) C82 and the Bayer combined yeast (Zygosaccharomyces bailii) Y4 are separated from the traditional black tea fungus and are preserved in the common microorganism center of China Committee for culture Collection of microorganisms with the preservation numbers of CGMCC No.16184 and CGMCC No.16183 respectively. Lactobacillus plantarum BC299 is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No. 6326.

3. Use according to claim 1 or 2, characterized in that: the medicine or food takes the bacterium colt C82, the Bayer combining yeast Y4 and the lactobacillus plantarum BC299 viable bacteria as effective components to prepare the dosage form which can be accepted in the medicine and/or food.

4. The use according to claim 1, characterized in that the black tea fungus is prepared by the following process:

1) selecting tea leaves: selecting dry green tea, black tea or other tea with strong tea flavor, wherein the tea mass percentage is 0.8%.

2) Leaching and filtering: adding folium Camelliae sinensis into boiling water, boiling for 5min, and filtering with edible filter cloth to obtain leaching solution.

3) And (3) sterilization: adding 4-10% white sugar into the filtered leaching solution, and sterilizing at 115 deg.C for 15 min.

4) Fermentation: mixing wood colt shaped bacteria C82, Bayer combining yeast Y4, Lactobacillus plantarum BC299 starter with water, and adding 10^6～8Inoculating CFU/100mL into sterilized tea soup, and fermenting at 30 deg.C for 4-7 days.

5) Centrifuging: the cells were removed by centrifugation at 8000r for 20min at 4 ℃.

6) Filling: and (3) mixing the centrifuged stock solution of the black tea fungus with 0-5 times of water, and filling to obtain the black tea fungus beverage.

5. Use according to claim 4, characterized in that: the purpose of extracting the tea in the step 2 is to extract tea polyphenol and other polyphenol components such as catechins, flavonoids, phenolic acids and the like in the tea into the tea soup.

6. Use according to claim 4, characterized in that: compared with the traditional black tea fungus, the black tea fungus beverage has the advantages that the content of short-chain fatty acids such as formic acid, acetic acid, butyric acid and the like is increased, the fasting blood glucose concentration is reduced, and the blood glucose reducing effect is increased.

7. The use according to claim 1, wherein the product functions are characterized in that: the important pathological characteristics of the type II diabetes mellitus comprise the relief of more than three and one less than three symptoms, the reduction of blood sugar concentration, the improvement of glucose tolerance and the improvement of the emaciation phenomenon of diabetic mice. Namely, the black tea fungus product can improve type II diabetes and improve the hypoglycemic effect by improving the weight of a mouse, reducing the food intake, the water intake and the urine output, reducing the fasting blood sugar and improving the glucose tolerance.

8. Use according to claim 7, characterized in that: the three-more-one-less comprises the increase of water drinking amount, the increase of food drinking amount, the increase of urine output and the decrease of body weight.

9. A medicine or food with hypoglycemic effect, which is characterized in that: the medicine or food comprises effective amount of black tea fungus and auxiliary materials acceptable in medicine and food.

10. The pharmaceutical or food product of claim 9, wherein: the dosage forms of the medicine or food comprise powder, tablets, granules, capsules and drinks.

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