Method for producing citron lemon ferment with the effect of increasing polyphenol intake and increasing adiponectin content

Abstract

The present application provides a method for producing a Citrus medica L. var. sarcodactylis fermentate with the effect of increasing polyphenol intake and adiponectin content. The method for producing a Citrus medica L. var. sarcodactylis fermentate with the effect of lowering blood lipids comprises the following steps: washing, air-drying, and breaking the cell wall, inoculating and fermenting, comprehensive filtering, sterilizing, and finally forming a Citrus medica L. var. sarcodactylis fermentate that can increase the total polyphenol content in the human body and improve the adiponectin content in the body.

Description

Method for producing fermented flat lemon that can increase polyphenol intake and adiponectin levels Technical Field

[0001] This invention relates to a method for producing a fermented flat lemon product; more specifically, it relates to a method for producing a fermented flat lemon product with lipid-lowering effects. Background Technology

[0002] In recent years, healthy eating has become a consensus pursued by the general public. Gyms and healthy restaurants have sprung up everywhere, reflecting people's pursuit of better physique and healthier diets. However, few people explore the essence of obesity, which stems from unhealthy diets and a lack of exercise. In addition to cultivating good habits in daily life, supplementing the body with the right energy sources, choosing foods high in polyphenols, and supplementing with hormones beneficial to the body, such as adiponectin, can all have a positive impact on health.

[0003] Polyphenols are protective substances produced by plants through photosynthesis after exposure to sunlight. They are widely found in the plant kingdom, second only to cellulose and lignin in content. Appropriate intake of polyphenols can bring many benefits to the human body. They can regulate physiological functions, reduce the burden caused by an unbalanced diet, and work synergistically with vitamin C to exert antioxidant effects. In the field of biotechnology, polyphenol content can also be used as an indicator of a plant's antioxidant capacity. The higher the polyphenol content, the stronger the antioxidant capacity. Moreover, consuming high levels of polyphenols can help regulate blood lipids, making it entirely beneficial to the human body.

[0004] Adiponectin is produced and secreted by adipocytes and is related to maintaining the metabolic balance of glucose and lipids in the body. Adiponectin has the functions of reducing triglyceride content in the liver and muscles, regulating blood lipid concentration, and regulating inflammatory response. Increasing the level of adiponectin in the blood can help regulate blood lipid content and lower blood lipid concentration.

[0005] Currently, there is a lack of products on the market that use flat lemons (also known as Taiwanese lemons) in combination with microbial fermentation to supplement the body with polyphenols and increase the body's adiponectin content.

[0006] In view of this, the inventor, based on his many years of research and development experience in related fields, conducted in-depth research on the aforementioned deficiencies and actively sought solutions to the aforementioned needs. After a long period of hard work and multiple tests, the inventor finally completed this invention. Summary of the Invention

[0007] The main objective of this invention is to provide a fermented flat lemon and its preparation method, wherein the fermented flat lemon can increase the daily intake of polyphenols and increase the content of adiponectin in the body, thereby achieving the effect of lowering blood lipids by combining the aforementioned two effects.

[0008] This invention discloses a method for producing a fermented product of *Lemon simonii* with lipid-lowering effects. The method is characterized by increasing the daily intake of polyphenols and simultaneously raising the level of adiponectin in the body, thereby achieving the lipid-lowering effect through the combination of these two effects. The method is prepared according to the following steps:

[0009] A. Cleaning, air drying and cell wall breaking lysis steps: Clean the surface of the flat lemon, air dry it, and then use a cell wall breaking device to break the cell wall to obtain flat lemon extract.

[0010] B. Fermentation Steps: The extract of *Citrus medica* is inoculated into the mixed bacterial solution and fermented at a temperature of 20℃~30℃ for 7~20 days to obtain *Citrus medica* fermentation broth. The ratio of the extract to the mixed bacterial solution is 5~20:80~95. The mixed bacterial solution consists of *Hanseniaspora uvarum* (BCRC 22779), *Saccharomyces bayanus* (BCRC 21816), *Lactobacillus bifermentans* (BCRC 12934), and *Lactobacillus plantarum* (BCRC 12251), with the ratio of the aforementioned mixed bacterial solution between 1~5:1~5:1~5:1~5.

[0011] C. Comprehensive filtration step: The fermented broth of flat lemons is filtered through a filtration device to remove impurities in preparation for the sterilization step.

[0012] D. Sterilization step: The fermented liquid of flat lemon is sterilized by passing it through a sterilization device to form the flat lemon ferment product with the lipid-lowering effect of the present invention.

[0013] The advantages of this invention's method for producing fermented flat lemon with lipid-lowering effects are:

[0014] 1. Increase total polyphenol intake: Fermentation using flat lemons and a combination of microorganisms increases the total daily intake of polyphenols.

[0015] 2. Increase the body's adiponectin content: By consuming the product of this invention, the body's adiponectin content will increase.

[0016] 3. By combining the two aforementioned effects, the effect of lowering blood lipids is achieved. Attached Figure Description

[0017] Figure 1 is a schematic diagram of a preferred embodiment of the preparation method of fermented flat lemon with lipid-lowering effect.

[0018] Figure 2 shows the experimental data of the preparation method of fermented flat lemon with lipid-lowering effect 1 - adiponectin content detection table in the blood of the test subjects. Detailed Implementation

[0019] In order to provide a more detailed understanding of the purpose, effects, features and structure of the present invention, preferred embodiments are described below with reference to the accompanying drawings.

[0020] Please refer to Figure 1, which is a schematic diagram of a preferred embodiment of the method for preparing the lipid-lowering fermented lemon product of the present invention.

[0021] As shown in Figure 1, the method for producing the lipid-lowering fermented lemon product of the present invention is as follows:

[0022] A. Cleaning, air drying and cell wall breaking lysis steps: Clean the surface of the flat lemon, air dry it, and then use a cell wall breaking device to break the cell wall to obtain flat lemon extract.

[0023] B. Fermentation Steps: The extract of *Citrus medica* is inoculated into the mixed bacterial solution and fermented at a temperature of 20℃~30℃ for 7~20 days to obtain *Citrus medica* fermentation broth. The ratio of the extract to the mixed bacterial solution is 5~20:80~95. The mixed bacterial solution consists of *Hanseniaspora uvarum* (BCRC 22779), *Saccharomyces bayanus* (BCRC 21816), *Lactobacillus bifermentans* (BCRC 12934), and *Lactobacillus plantarum* (BCRC 12251), with the ratio of the aforementioned mixed bacterial solution between 1~5:1~5:1~5:1~5.

[0024] C. Comprehensive filtration step: The fermented broth of flat lemons is filtered through a filtration device to remove impurities in preparation for the sterilization step.

[0025] D. Sterilization step: The fermented liquid of flat lemon is sterilized by passing it through a sterilization device to form the flat lemon ferment product with the blood lipid lowering effect of the present invention.

[0026] In the aforementioned inoculation step, the selection of strains is aimed at maximizing the polyphenol content of the finished product, allowing consumers to ingest the maximum amount of polyphenols, thereby further enhancing the benefits provided to the human body by this invention.

[0027] Continuing from the foregoing, the selection of this strain has been demonstrated by the following multiple examples to have a better mixed strain, resulting in a higher total polyphenol content in the final fermented flatbread lemon product of the present invention.

[0028] In several embodiments, the extract of *Citrus medica* was inoculated into a mixed bacterial culture of different strains and fermented at a temperature of 20°C to 30°C for 7 to 20 days to obtain *Citrus medica* fermentation broth. The ratio of the extract of *Citrus medica* to the mixed bacterial culture was 5 to 20: 80 to 95. After completion, the polyphenol content of each strain was measured, and the strain with the highest total polyphenol content was selected as the preferred embodiment.

[0029] The following table and figures present several embodiments to illustrate the technical effects achieved by the method provided by the present invention; please refer to Table (I), which is a table of the names and numbers of all bacterial species used in the following embodiments.

[0030] Strain Number | Strain Name | 1. Hanseniaspora uvarum (BCRC 22779) | 2. Saccharomyces bayanus (BCRC 21816) | 3. Lactobacillus bifermentans (BCRC 12934) | 4. Lactobacillus plantarum (BCRC 12251) | 5. Lactobacillus brevis (BCRC 10361) | 6. Bifidobacterium bifidum (BCRC 12585) | 7. Lactobacillus casei (BCRC 10697) surface

[0031] Table (1)

[0032] Example 1

[0033] The fermentation liquid of the flat lemon was prepared by fermentation according to the method for preparing the flat lemon fermentation product with the effect of lowering blood lipids. However, the mixed bacterial liquid was composed of numbers 1, 2, 3 and 4. In the bacterial fermentation step, the temperature was 20°C and the fermentation was carried out for 20 days. The ratio of the flat lemon extract to the mixed bacterial liquid was 20:80 and the ratio of the mixed bacterial liquid was 1:1:1:5.

[0034] Example 2

[0035] The fermentation liquid of the flat lemon was prepared by fermentation according to the method for preparing the flat lemon fermentation product with the effect of lowering blood lipids. However, the mixed bacterial liquid was composed of numbers 1, 2, 3 and 4. In the bacterial fermentation step, the temperature was 25°C and the fermentation time was 14 days. The ratio of the flat lemon extract to the mixed bacterial liquid was 15:85 and the ratio of the mixed bacterial liquid was 1:5:1:5.

[0036] Example 3

[0037] The fermentation liquid of the flat lemon was prepared by fermentation according to the method for preparing the flat lemon fermentation product with the effect of lowering blood lipids. However, the mixed bacterial liquid was composed of numbers 1, 2, 3 and 4. In the bacterial fermentation step, the temperature was 28°C and the fermentation was carried out for 10 days. The ratio of the flat lemon extract to the mixed bacterial liquid was 10:90 and the ratio of the mixed bacterial liquid was 5:1:5:1.

[0038] Example 4

[0039] The fermentation liquid of the flat lemon was prepared by fermentation according to the method for preparing the flat lemon fermentation product with the effect of lowering blood lipids. However, the mixed bacterial liquid was composed of numbers 1, 2, 3 and 4. In the bacterial fermentation step, the temperature was 30°C and the fermentation was carried out for 7 days. The ratio of the flat lemon extract to the mixed bacterial liquid was 5:95 and the ratio of the mixed bacterial liquid was 5:5:5:1.

[0040] Comparative Example 1

[0041] The fermentation liquid of the flat lemon was prepared by fermentation according to the method described in Example 1, except that the ratio of the mixed bacterial liquid was 1:0.5:0.5:5.

[0042] Comparative Example 2

[0043] The fermentation liquid of the flat lemon was prepared by fermentation according to the method described in Example 2, except that the ratio of the mixed bacterial liquid was 1:6:1:6.

[0044] Comparative Example 3

[0045] The fermentation liquid of the flat lemon was prepared by fermentation according to the method described in Example 3, except that the ratio of the mixed bacterial liquid was 5:0.5:5:0.5.

[0046] Comparative Example 4

[0047] The fermentation liquid of the flat lemon was prepared by fermentation according to the method described in Example 4, except that the ratio of the mixed bacterial liquid was 6:6:6:1.

[0048]

[0049] Table (II)

[0050] Please refer to Table (II), which summarizes the changes in total polyphenol content in the lipid-lowering fermented products of *Lemon strumarium* obtained in Examples 1 to 4 and Comparative Examples 1 to 4. As can be seen from the above experiments, under the mixed strain ratio conditions of Examples 1 to 4, the total polyphenol content of *Lemon strumarium* fermented products was effectively increased. On the other hand, under the conditions of excessively high or low proportions of individual strains in Comparative Examples 1 to 4, the total polyphenol content of *Lemon strumarium* fermented products could not be effectively increased, and under certain conditions, the total polyphenol content in *Lemon strumarium* fermented products was even reduced.

[0051] Example 5

[0052] The fermentation broth of the flat lemon was prepared by fermentation according to the method of Example 2, except that the mixed bacterial broth was classified into groups A, B, C, D and E respectively; please refer to Table (III), which shows the group number, the bacterial strains used and the composition ratio of each group. Group A consists of numbers 1, 2, 3 and 4, group B consists of numbers 1, 3, 4 and 6, group C consists of numbers 2, 4, 5 and 7, group D consists of numbers 1, 2, 5 and 6, and group E consists of numbers 2, 3, 4 and 7.

[0053]

[0054] Table (III)

[0055] As can be seen from the above experiments, the mixed bacteria in group A, after being fermented with the extract of flat lemon, can produce the largest amount of total polyphenols. Therefore, this group was used as the inoculum for the mixed bacterial liquid of the present invention for fermentation, so that the final flat lemon fermented product also has a better total polyphenol content. When the human body ingests the flat lemon fermented product with a higher content of total polyphenols, it can supplement the various components in the excellent total polyphenols, thereby improving the nutrients required by the human body every day.

[0056] Example 6

[0057] Continuing from the foregoing, the fermented flat lemon of this invention, which has the effect of lowering blood lipids, can increase the concentration of adiponectin in the blood and break down greasy fats in the diet. Its efficacy can be verified through the following experiments.

[0058] First, C57BL / 6HBV-Tg mice were used as test subjects in this experiment. They were fed a high-fat diet and a high-fat diet with fermented lemon sylvestris var. chinensis added to the diet of this invention. 1 to 8 grams of fermented lemon sylvestris var. chinensis added to 1 kg of the high-fat diet were obtained from Example 2. The experiment lasted for 12 weeks. At the end of the experiment, blood samples were drawn from the mice for testing. The aforementioned H NMR spectroscopy was used to analyze the mouse blood samples to detect the adiponectin content in the blood samples. The following test results were obtained to demonstrate the efficacy of the fermented lemon sylvestris var. chinensis var. chinensis in lowering blood lipids.

[0059] Please refer to Figure 2. Figure 2 shows the experimental data of the preparation method of fermented flat lemon with lipid-lowering effect - adiponectin content detection table in the blood of the test subjects.

[0060] As shown in Figure 2, after the 12-week experiment, among the subjects fed two different diets (high-fat diet and high-fat diet with fermented lemon syrup), the adiponectin content in the blood of the subjects fed the high-fat diet with fermented lemon syrup was significantly higher than that of the subjects fed the high-fat diet. It can be inferred that if the human body ingests the product with fermented lemon syrup, it can increase adiponectin levels, thereby reducing the triglyceride content in the liver and muscles, and indirectly achieving the function of regulating blood fat concentration and regulating inflammatory response.

[0061] In summary, the advantages of the fermented flat lemon product of the present invention, which has the effect of lowering blood lipids, are as follows:

[0062] 1. Increase daily polyphenol intake: Fermentation using flat lemons and paired bacteria provides additional high-quality polyphenols, increasing the body's daily polyphenol intake.

[0063] 2. Increase the body's adiponectin content: By consuming the product of this invention, the body's adiponectin content will increase.

[0064] 3. Lowering blood lipids and improving health: Ingesting high levels of polyphenols and increasing adiponectin levels can help regulate blood lipids and thus lower blood lipid levels.

[0065] Therefore, this invention has excellent progressiveness and practicality among similar products. After reviewing all technical data and literature on this type of structure at home and abroad, no identical or similar structures were found prior to this application. Therefore, this application should meet the patent grant conditions of novelty, inventiveness and practicality, and is therefore filed in accordance with the law.

[0066] However, the above description is only a preferred embodiment of the present invention. Any other equivalent structural changes made in accordance with the present invention specification and the scope of patent protection should be included within the scope of patent protection of the present invention.

Claims

1. A method for producing a fermented flat lemon product that increases polyphenol uptake and adiponectin content, characterized in that, The process includes the following steps: A. Cleaning, air drying and cell wall breaking and lysis steps: The surface of the flat lemon is cleaned and air dried, and then cell wall breaking and lysis are carried out using a cell wall breaking device to obtain flat lemon extract. B. Fermentation Steps: The extract of *Lysimachia christinae* is inoculated into the mixed bacterial solution and fermented at 20℃~30℃ for 7~20 days to obtain *Lysimachia christinae* fermentation broth. The ratio of the *Lysimachia christinae* extract to the mixed bacterial solution is 5~20:80~95. The mixed bacterial solution is derived from the following strains: *Hanseniaspora uvarum* (BCRC 22779), *Saccharomyces bayanus* (BCRC 21816), *Lactobacillus bifermentans* (BCRC 12934), and *Lactobacillus plantarum* (BCRC 2000). 12251 is composed of ingredients in ratios of 1:1:1:5, 1:5:1:5, 5:1:5:1, or 5:5:5:1; C. Comprehensive filtration step: The fermented liquid of flat lemon is filtered through a filtration device to remove impurities in preparation for sterilization; and D. Sterilization step: The fermented liquid of flat lemon is sterilized through a sterilization device to form a fermented product of flat lemon that has the effect of increasing polyphenol uptake and increasing adiponectin content.

2. The method for producing the fermented flat lemon product with the effects of increasing polyphenol uptake and increasing adiponectin content according to claim 1, characterized in that, The cleaning equipment used in the washing, drying, and cell wall breaking processes can clean the surface of the flat lemons through aeration or ultrasonic waves.

3. The method for producing the fermented flat lemon product with the effects of increasing polyphenol uptake and increasing adiponectin content according to claim 1, characterized in that, The washing, drying, and cell wall breaking processes are carried out by physical cell wall breaking equipment to break down the flat lemons.

4. The method for preparing fermented flat lemon according to any one of claims 1 to 3, characterized in that, In the bacterial fermentation step, the temperature is 25℃~28℃.

5. The method for preparing the fermented flat lemon product according to any one of claims 1 to 3, characterized in that, In the aforementioned bacterial fermentation step, fermentation is carried out for 10 to 14 days.

6. The method for preparing the fermented flat lemon product according to any one of claims 1 to 3, characterized in that, In the bacterial fermentation step, the ratio of the flat lemon extract to the mixed bacterial solution is 10~15:85~90.

7. The method for preparing fermented flat lemon according to any one of claims 1 to 3, characterized in that, In the aforementioned bacterial fermentation step, the temperature is 25℃~28℃, and the fermentation time is 10~14 days, wherein the ratio of the flat lemon extract to the mixed bacterial solution is 10~15:85~90.

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