TWI640316B - Microbial fermented product through music fermentation, manufacturing method and use thereof - Google Patents

Abstract

The invention provides a microbial ferment, which is a vegetable juice and berry juice mixed liquid obtained by fermenting Streptococcus thermophilus, yeast and acetic acid bacteria at a frequency; wherein the frequency is in classical music, pop music or Rock and roll. The microbial fermentation product of the present invention promotes fat cell metabolism and protects the liver.

Description

   Microbial fermentation product cultured under music environment, preparation method and application thereof   

The invention provides a microbial fermentation cultured in a music environment, and particularly relates to a microbial fermentation cultured in a music environment and capable of promoting fat cell metabolism and protecting the liver.

For a long time, obesity has been defined as a disease condition that is associated with most health problems and with reduced lifespan. In the modern era of people striving for good health, various nutrition and slimming supplements are gradually appearing on the market to meet people's needs; however, most of these nutritional and slimming supplements are chemically synthesized or are not found in natural foods. The combination of a single molecular structure, the raw material sources are petroleum extracts, coal pitch derivatives, sugar after chemical processing or industrialized products after processing, but these chemically extracted nutrients are difficult to be recognized by cells, Accept and use, so more nutrients are actively excreted with urine or sweat. Due to the low absorption rate, low dwell time, and low bioavailability of these nutritional supplements, chemically synthesized nutrients have not reached their declared efficacy, and the content may be hundreds or even thousands of times the recommended daily intake. High doses will have May bring the risk of toxic reactions to the human body; Although the body will actively excrete excess nutrients, it also means that unabsorbed chemical nutrients need liver and kidney metabolism, which will increase the extra burden on the human body; and long-term consumption of chemically synthesized nutrients may accumulate Toxic with side effects and even carcinogenic risk.

Therefore, how to use the nutritional ingredients made from natural foods does not require major changes in lifestyle and does not cause toxins or adversely affect health. At the same time, it can achieve the dual effects of health care and reducing obesity. It is the goal that people strive for for a long time.

In view of this, it is an object of the present invention to provide a microbial fermented product, which is a vegetable juice and berry juice mixed liquid with a Streptococcus thermophilus , a yeast and an acetic acid bacteria at a frequency Obtained by fermentation; where the frequency is less than 10,000Hz within 1 to 3 minutes of classical music, accounting for more than 99% of the entire song, and less than 10,000Hz within 1 to 3 minutes of popular music, accounting for 95% of the entire song % Or more, or more than 10,000Hz within 1 to 3 minutes of a rock track, account for more than 50% of the entire track.

Another object of the present invention is the use of the microbial fermentation product for preparing a pharmaceutical composition that promotes fat cell metabolism, wherein the microbial fermentation product is less than 10,000 Hz in the popular music track within 1 to 3 minutes and accounts for the entire song. It is obtained by fermentation at a frequency of more than 95% of the tracks.

Another object of the present invention is the use of the microbial fermented material for preparing a pharmaceutical composition for protecting the liver, wherein the microbial fermented material is less than 10,000 Hz and accounts for the entire song within 1 to 3 minutes of classical music. It is obtained by fermentation at a frequency of more than 99%.

In one embodiment of the present invention, the S. thermophilus strain is a strain of BCRC 910636, the yeast strain is a strain of Saccharomyces cerevisiae BCRC 21494, and the acetic acid strain is Acetobacter aceti ) BCRC12324 or BCRC11688 strains.

In one embodiment of the present invention, the fat cell metabolism system is to promote lipolysis or accelerate brown fat cell decomposition.

In one embodiment of the present invention, the promotion of lipolysis comprises increasing the gene expression of adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (LIPE); and Accelerating the decomposition of brown adipocytes includes increasing the gene expression of uncoupling protein 1 (UCP1) and adipose uncoupling protein 2 (UCP2).

In one embodiment of the present invention, the liver protection system is to improve a liver cell's anti-oxidation ability or a DNA repair ability, and the DNA repair ability is to improve a DNA structure damage or a DNA double strand break repair ability.

In one embodiment of the present invention, increasing the antioxidant capacity of the liver cells includes increasing the gene expression of superoxide dismutase 1 (SOD1) and superoxide dismutase 2 (SOD2), and increasing the DNA. The ability to repair structural damage includes increasing the gene expression of excision repair cross-complementing gene 1 (ERCC1) and DNA damage recognition and repair factor (XPA), and increasing the DNA The repair ability of double-strand breaks includes increasing the gene expression of X-ray repair cross complementation 1 (XRCC1) and X-ray repair cross complementation 2 (XRCC2).

In one embodiment of the present invention, the total volume of the Streptococcus thermophilus, the yeast and the acetic acid bacteria is 2 to 7% of the volume of the vegetable juice and berry juice mixed liquid.

The embodiments of the present invention will be further described below with reference to the drawings. The examples listed below are intended to clarify the present invention and are not intended to limit the scope of the present invention. Any person skilled in the art will not depart from the spirit and scope of the present invention. Within the scope, when some changes and retouching can be done, the protection scope of the present invention shall be determined by the scope of the attached patent application scope.

FIG. 1 is a superoxide dismutase-like (SOD-like) content of a microbial fermentation cultured in a music environment according to the present invention.

FIG. 2 is a data chart showing the relative expression of the superoxide dismutase 2 (SOD2) gene of gastric epithelial cells cultured by a microbial fermentation in a musical environment according to the present invention.

FIG. 3 is a data chart showing the relative expression levels of the superoxide dismutase 1 (SOD1) and superoxide dismutase 2 (SOD2) genes of oxidatively damaged liver cells cultured by microbial fermentation in a music environment according to the present invention.

FIG. 4 is the excision repair cross-complementing gene 1 (ERCC1) and DNA damage recognition and repair factor (ERCC1) of oxidative-damaged hepatocytes cultured by a microbial fermentation in a musical environment according to the present invention Repair factor (XPA) gene relative expression data.

FIG. 5 shows X-ray repair cross complementing 1 (XRCC1) and X-ray repair cross-complementing gene 2 (XRCC2) of oxidative-damaged hepatocytes cultured by microbial fermentation in a music environment according to the present invention Data of relative gene expression.

FIG. 6 is a data chart of oil-red O stain analysis of fat metabolism by microbial fermentation cultured in a music environment according to the present invention.

FIG. 7 is a data chart of relative expression amounts of adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (LIPE) genes of microbial fermentation cultured in a music environment according to the present invention .

FIG. 8 is a data chart of the relative expression of the uncoupling protein 1 (UCP1) and the fat uncoupling protein 2 (UCP2) genes of the microbial fermentation cultured in a music environment according to the present invention.

The invention uses a liquid as a medium for sound transmission, and accelerates the intercellular reaction at a speed of sound (approximately 1463 m / s) that the microorganisms can hear during fermentation. After sonic stimulation, the activity of microorganisms is significantly improved, which accelerates the metabolism, synthesis, oxidation and reduction capabilities. It is because the sonic species' alternating stress field causes a slight change in the structure of the microorganisms, and enhances the fluidity and permeability of nutrients in the cells (Ca ²⁺ , Mg ^2+, etc.), resulting in The change of ion concentration inside and outside the membrane causes the directional migration of endogenous auxin and related growth-influencing substances in cells, thereby changing the ability of cells to absorb and operate substances. Therefore, in the present invention, by playing music during the fermentation process of the mixed liquid of vegetable juice and berry juice, the fermented matter can receive energy of different frequencies and wavelengths in the music to generate a microbial fermented matter. At the same time, the present invention measures the superoxide dismutase-like (SOD-like) content of the microbial fermentation, the antioxidative power of gastric cells, the antioxidative power of liver cells, the ability of DNA repair of liver cells, and Promote fat cell metabolism.

definition

The vegetables described herein include, but are not limited to, vegetables such as leafy vegetables, bean sprouts, fruits and vegetables, solanaceous fruits, potatoes, root vegetables, cauliflowers, and fruits such as stone fruits, kernels, and melons.

The berries described herein include, but are not limited to, blueberries, cranberries, red grapes, white grapes, bayberry, mulberry, apples, carrots, sugar cane, passion fruit, pineapple, and lemon.

Statistical analysis was performed using Excel software. The data are expressed as mean ± standard deviation (SD), and the differences between them are analyzed by student's t-test.

As the values used herein are approximate, all experimental data are shown in the range of 20%, preferably in the range of 10%, and most preferably in the range of 5%.

Example 1 A method for fermenting a microorganism cultured in a musical environment according to the present invention

In a preferred embodiment of the present invention, the vegetable juice and the berry juice are mixed at a ratio of 3 to 2, and then diluted with 5 times the volume of distilled water of the vegetable juice and the berry juice as an empty culture liquid; wherein the berry juice contains: Blueberry, cranberry, red grape, white grape, bayberry, mulberry, apple, carrot, sugar cane, passion fruit, pineapple, and lemon are mixed in equal proportions; vegetable juice contains: broccoli, celery and asparagus, mixed in equal proportions. The aerial culture solution was pasteurized, and then connected to Streptococcus thermophilus TCI 633 (Institute of Food Industry Development, No. BCRC 910636, Republic of China Patent Publication No. I519644 Patent Deposit), Saccharomyces cerevisiae ( Saccharomyces cerevisiae ) BCRC 21494 and Acetobacter aceti BCRC12324 or BCRC11688 into the air culture solution, and pre-fermentation is performed under the condition of constant temperature fermentation at 30 ° C. ± 1 ° C. for 192 hours to obtain the microorganism fermentation product of the present invention, among which these strains The total volume is 2 to 7% of the volume of the air culture solution; it is divided into three groups during the fermentation process, and three types of music are played, divided into classical music (Mozart: Symphony No. 40-First Movement), pop music (Maroon 5: sugar) and rock music (Maroon 5: Maps), among which classical music is less than 10,000Hz within 1 to 3 minutes, accounting for more than 99% of the entire song, and popular music is songs 1 to 3 Less than 10,000Hz accounted for more than 95% of the entire track in minutes, and rock music was more than 10,000Hz accounted for more than 50% of the entire track within 1 to 3 minutes.

Example 2 Detection of Superoxide Dismutase (SOD-like) Content Like the Microbial Fermentate of the Present Invention

Superoxide dismutase (SOD) is an enzyme that catalyzes the dismutation of superoxide radicals into oxygen and hydrogen peroxide. These enzymes have an important anti-oxidant defense effect in cells exposed to oxygen. Therefore, the present invention uses a SOD activity kit (SOD activity kit, Enzo Life Science Company, model CAT # ADI-900-157) for cell oxidative stress analysis.

The results are shown in Figure 1. Compared with the control group (only enzymes that have not listened to music), microbial fermentates cultured in any music environment (classical music, pop music and rock music) can effectively improve superoxide-like compounds. The dismutase (SOD-like) content is more than 1 time, and it can be inferred that its antioxidant capacity will be improved than that of microbial fermentations not cultured in a musical environment. The results show that the microbial fermented material of the present invention contains a higher activity of superoxide dismutase, which can have a stronger defense against oxidative stress.

Example 3 Antioxidant Capability of Gastric Cells of the Microbial Fermentate of the Present Invention

In the present invention, the cell platform is used to test the antioxidant capacity, and quantitative real-time polymerase chain reaction (Quantitative real time PCR, qPCR) is used to detect the superoxidation of AGS human gastric epithelial cells treated with the microbial fermentation of the present invention. The gene expression of the substance dismutase 2 (SOD2) was used to analyze the antioxidative capacity of gastric cells of the microbial fermented product of the present invention.

In the present invention, human gastric epithelial cells were purchased from the American Type Culture Collection (ATCC) and numbered CRL-1739. The cells were cultured with the addition of 10% fetal bovine serum (FBS) (GIBCO, number 10438-026, USA) and 1% penicillin / streptomycin (GIBCO, number 15140-122). RPMI 1640 (Roswell Park Memorial Institute) (GIBCO, USA).

1.5 × 10 ⁵ human gastric epithelial cells were inoculated into each well of a 6-well culture plate (GeneDireX, Taiwan), and cultured in the above-mentioned medium at 37 ° C. in a 5% CO ₂ incubator for 24 hours; without disturbing the attachment Remove the medium in the case of cells on the culture plate, and then replace the new medium with 2% concentration of the microbial fermentation of the present invention under three different music environments for 6 hours, and control the cells containing only the medium as control Group, the same experimental conditions described above were performed in triplicate. Human gastric epithelial cells were then collected. RNA was extracted from the above cells using an RNA extraction reagent kit (Geneaid, Taiwan), and then reverse-transcribed into cDNA using reverse transcriptase (SuperScript® III Reverse Transcriptase) (Invitrogen, USA), followed by KAPA SYBR® FAST qPCR reagent Group (KAPA Biosystems, USA) performed quantitative real-time polymerase chain reaction (qPCR) with an instant polymerase chain reaction system (ABI StepOne Plus ^TM System (Applied Biosystems, USA)) to detect the target gene (SOD2) and the reference gene The expression level of the target gene is standardized by the expression level of the reference gene.

The results are shown in Figure 2. Compared with cells containing only culture medium as the control group, no matter which music environment (classical music, pop music, and rock music) cultured gastric epithelial cells treated with microbial fermentation, its antioxidant-related SOD2 Gene expression can be effectively increased by more than 20%.

Example 4 Antioxidant ability of hepatocytes of the microbial fermentation product of the present invention

The present invention uses hydrogen peroxide (H ₂ O ₂ ) to induce oxidative damage to liver cells. Since it is known that superoxide dismutase (SOD) can catalyze superoxide and reduce the content of free radicals, it is an important antioxidant in the human body. ; The present invention uses quantitative real-time polymerase chain reaction (qPCR) to detect the gene expression of superoxide dismutase 1 (SOD1) and superoxide dismutase 2 (SOD2) of oxidatively damaged liver cells treated with the microbial fermentation product of the present invention. The anti-oxidative capacity of the hepatocytes of the microbial fermentation product of the present invention was analyzed.

In the present invention, HepG2 liver cells (liver hepatocellular cells) were purchased from the American Standard Biological Collection Center (ATCC) and numbered HB-8065. Hepatocyte culture conditions were the same as described in Example 1, and 4mL H ₂ O _{2 was used to} induce oxidative damage to the cells. Then, 2% concentrations of three different microbial fermentations of the present invention in a different music environment were added for 6 hours. Quantitative real-time polymerase chain reaction (qPCR) was used to detect the expression levels of SOD1 and SOD2 genes in oxidatively damaged liver cells treated with the microbial fermentation product of the present invention. The detection method was as described in Example 3. The same experimental conditions described above were performed in triplicate.

The results are shown in Fig. 3. Compared to cells containing only culture medium as the control group and cells induced only with H ₂ O ₂ induced oxidative damage, SOD1 of cultured hepatocytes treated with microbial fermentate was cultured in classical music and pop music environments. The expression levels of both SOD2 and SOD2 genes were significantly increased. At the same time, cells treated with H ₂ O ₂ induced oxidative damage and treated with the empty culture solution of Example 1 as a control group could also increase the expression of SOD1 and SOD2 genes in oxidatively damaged liver cells. However, the expression of SOD1 and SOD2 genes in hepatocytes treated with microbial fermentation in the classical music environment was the best. It is shown that the microbial fermented material of the present invention can improve the oxidizing ability of hepatocytes, and also has quite good effects on protecting hepatocytes.

Example 5 Hepatocyte DNA repair ability of the microbial fermentation product of the present invention

Because oxidatively damaged cells are known to produce free radicals, free radicals can cause DNA damage and gene mutations. In the present invention, oxidative damage to HepG2 liver cells is induced by hydrogen peroxide (H ₂ O ₂ ), and the microorganism fermentation product of the present invention under three different music environments is added at a concentration of 2%, respectively. After 6 hours of treatment, quantitative instant polymerase chain reaction is performed. (qPCR) Detects the DNA structure damage of oxidatively-damaged hepatocytes treated with the microbial fermentation of the present invention and the expression of repair genes for double-stranded DNA breaks; wherein the DNA structure damage repair genes include the excision repair cross gene 1 (excision repair cross -complementing gene 1 (ERCC1) and DNA damage recognition and repair factor (XPA); repair genes for double-stranded DNA breaks include X-ray repair cross complementation 1 (XRCC1) ) And X-ray repair cross-complementary gene 2 (XRCC2), the detection method is as described in Example 3, and the same experimental conditions described above are performed in triplicate.

The results are shown in Fig. 4 and Fig. 5. Compared with cells containing only culture medium as the control group and cells induced with oxidative damage only by H ₂ O ₂ , no matter what the music environment (classical music, pop music, and rock music) The expression of ERCC1, XPA, XRCC1 and XRCC2 genes in cultured oxidatively damaged liver cells treated with fermented microorganisms were significantly increased. At the same time, cells treated with H ₂ O ₂ induced oxidative damage and treated with the unlistened music enzyme product control group could improve Expression of ERCC1 and XPA genes in oxidatively damaged hepatocytes (Figure 4); cells induced with H ₂ O ₂ and treated with the empty culture solution of Example 1 can also enhance the XRCC1 and XRCC2 genes of oxidatively damaged liver cells The expression level (Figure 5), but the average expression level of ERCC1, XPA, XRCC1, and XRCC2 genes of hepatocytes treated with microbial fermentation in a classical music environment is the best. It has been shown that the microbial fermentation product of the present invention can improve the repair ability of DNA damage of liver cells (ERCC1 and XPA) and the gene expression of double-stranded DNA (XRCC1 and XRCC5) also has the ability to repair, and it is also quite good for protecting liver cells. Effect.

Example 6 Fatty Cell Metabolism Promoting Capability of the Microbial Fermentate of the Present Invention

In the present invention, dexamethasone (DEXA) and isobutylmethylxanthine (IBMX) are used to induce mouse 3T3-L1 preadipocytes to differentiate into mature adipocytes, and then add three different kinds of music separately After the microorganism fermentation product of the present invention is treated in the environment, oil-red O staining is used to confirm the effect of promoting fat cell decomposition by the microorganism fermentation product of the present invention, and quantitative real-time polymerase chain reaction (qPCR) detection The expression amount of the fat metabolism gene treated by the microbial fermentation product of the present invention.

6.1 3T3-L1 cell experimental analysis method

First, place 3T3-L1 pre-adipocytes in each well of a 6-well cell culture plate and inoculate 1.5 × 10 ⁵ cells. After the cells are cultured to be dense, it is regarded as the zero day. At this time, 10% fetal bovine serum and 10 μg are added. / ml insulin, 1 μM DEXA, and 0.5 mM IBMX dulbecco's modified eagle medium (DMEM), cultured for 48 hours; on the third day, the medium was replaced (using the aforementioned medium), and cultured for 48 hours; on the fifth day, the medium was added and replaced DMEM medium with 10% fetal bovine serum and 10 μg / mL insulin. Change the medium every two days and continue to differentiate to day 6. When the cells begin to differentiate into adipocytes, add three different music environments at 1% or 2% concentration, respectively. The microorganism fermentation product of the present invention was put into each sample, and the observation was continued for 8 days; Oil red-O stain was performed, and the oil red stain was a specific stain for neutral fat cells; the medium was aspirated , Washed twice with PBS and fixed the cells with 10% Formaldehyde for 30 minutes; washed the cells twice with PBS and then stained with oil red O dye (dissolved in isopropanol) for one hour, and finally de-stained with PBS , You can Microscopically; stained with 100% isopropanol dissolve grease, and in a wavelength measured absorbance 510nm (OD).

In the present invention, oil-red O staining is used to confirm the fat cell decomposition promoting effect of the microbial fermentation product of the present invention. The results are shown in FIG. 6. In the fat metabolism experiment, compared with the fat cells containing only the culture medium as the control group, the fat cells treated with the 2% air culture solution of Example 1 and the 1% or 2% pandanase Treated fat cells, whether cultured in a 1% or 2% music environment (classical music, pop music, and rock music), cultured microbial fermented fat cells have the ability to significantly promote lipolysis, and the higher the concentration of microbial fermented matter The better the effect.

6.2 Analysis of gene expression in adipocytes

Put 3T3L1 and 3T3-L1 preadipocytes in each well of a 6-well cell culture plate to inoculate 2 × 10 ⁵ cells. When the cells are cultured to be dense, it is regarded as the zero day. At this time, 10% fetal bovine serum, 10 μg are added. / ml of insulin, 1 μM dexamethasone (DEXA), and Dow's modified Eagle's medium (DMEM) with 0.5 mM isobutylmethylxanthine (IBMX), cultured for 48 hours; on the third day, the medium was replaced (using the aforementioned medium), and cultured for 48 hours On the fifth day, the medium was replaced with DMEM medium supplemented with 10% fetal bovine serum and 10 μg / mL insulin. The medium was changed every two days and continued to differentiate to day 6. When the cells began to differentiate into adipocytes, they were added to three different music environments. The microorganism fermentation product of the present invention was processed into each sample for 24 hours; cells were collected. RNA was extracted from the above cells using an RNA extraction reagent kit (Geneaid, Taiwan), and then reverse-transcribed into cDNA using reverse transcriptase (SuperScript® III Reverse Transcriptase) (Invitrogen, USA), followed by KAPA SYBR® FAST qPCR reagent The group (KAPA Biosystems, USA) performed quantitative real-time polymerase chain reaction (qPCR) with an instant polymerase chain reaction system (ABI StepOne Plus ^TM System (Applied Biosystems, USA)) to detect the treatment of the microbial fermentation of the present invention. Expression of fat metabolism genes; wherein the fat metabolism genes include adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (LIPE) genes that promote lipolysis; and accelerate brown fat cells Decomposition includes promotion of uncoupling protein 1 (UCP1) and adipose uncoupling protein 2 (UCP2) genes. The detection method is as described in Example 3. The same experimental conditions described above are performed in triplicate.

The results are shown in Figs. 7 and 8. Compared with the adipocytes containing only culture medium as the control group and the adipocytes treated with the 2% air culture solution of Example 1, no matter what the music environment (classical music, pop (Music and rock music) the expression of ATGL and LIPE genes in cultured microbial fermented fat cells has increased significantly, indicating that the microbial fermented material of the present invention does have the ability to promote lipolysis; and pop music cultured microbial fermented fat cells The UCP1 and UCP2 genes have the best expression, indicating that the microbial fermented product of the present invention does have the effect of promoting the decomposition of brown fat.

In summary, the invention uses different types of music during the fermentation process of the mixed liquid of vegetable juice and berry juice, so that the fermented matter can receive the energy of different frequencies and wavelengths in the music to produce the microbial fermented matter. In the fermentation process, no matter what kind of music is given (classical music, pop music, or rock music), the microbial fermentation product of the present invention can effectively increase the content of superoxide dismutase-like (SOD-like) components by more than 1 time, and it can be inferred that In terms of anti-oxidation ability, it will be more improved than the microbial fermentation which is not cultured in a music environment. The present invention further uses the cell platform to test the anti-oxidation ability. The gastric epithelial cells are treated with the microbial ferment of the present invention under three different music environments at a concentration of 2%, and then SOD2 is detected by quantitative real-time polymerase chain reaction (qPCR) Gene expression, and the results show that the microbial fermentation product of the present invention has the effect of increasing the expression of the antioxidant SOD2 gene.

In addition, the microbial ferment of the present invention also has excellent effects on protecting the liver. The expression of SOD1 and SOD2 genes of hepatocytes treated with microbial ferment in a classical music environment is increased, and further study of the microbial ferment can also improve DNA repair ability, to improve the performance of DNA structural damage repair genes (ERCC1 and XPA) and double-strand DNA break repair genes (XRCC1 and XRCC5).

Furthermore, in the experiments to promote the metabolism of fat cells, the fat cells treated with the microbial fermentate of the present invention at a concentration of 1% and 2% have the ability to promote lipolysis, and the microbial fermentate-treated fat is cultured in a pop music environment. The increase in the expression levels of fat metabolism genes (ATGL and LIPE) and accelerated brown fat cell decomposition genes (UCP1 and UCP2) in cells indicates that the microbial fermentation product of the present invention has the effect of promoting the decomposition of brown fat.

Claims (12)

A microbial fermentation product is obtained by fermenting a mixture of vegetable juice and berry juice with a Streptococcus thermophilus , a yeast, and an acetic acid bacteria at a frequency; wherein the frequency is a classical Tracks below 10,000Hz account for more than 99% of the entire track within 1 to 3 minutes, Tracks below 10,000Hz account for more than 95% of the entire track within 1 to 3 minutes, or Rock Tracks for 1 to 3 minutes The higher than 10,000 Hz accounted for more than 50% of the entire track; the Streptococcus thermophilus strain was a strain of BCRC 910636. The microbial fermentation product according to item 1 of the scope of the patent application, wherein the yeast strain is a strain of Saccharomyces cerevisiae BCRC 21494. The microbial fermentation product according to item 1 of the scope of the patent application, wherein the acetic acid strain is an Acetobacter aceti BCRC12324 or a BCRC11688 strain. The use of the microbial fermentate as described in item 1 of the scope of the patent application for the preparation of a pharmaceutical composition for promoting lipolysis, wherein the microbial ferment is less than 10,000 Hz in the popular music track within 1 to 3 minutes and accounts for the entire track Obtained by fermentation at a frequency of more than 95%. The use as described in item 4 of the scope of patent application, wherein the promotion of lipolysis comprises increasing the gene expression of adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (LIPE) . The use as described in item 4 of the scope of patent application, wherein the lipolysis promoting system is to accelerate the decomposition of brown adipocytes, and the accelerated decomposition of brown adipocytes comprises increasing uncoupling protein 1 (UCP1) and fat uncoupling Gene expression of protein 2 (UCP2). The use of the microbial fermentate as described in item 1 of the scope of the patent application for the preparation of a pharmaceutical composition for protecting the liver, wherein the microbial fermentate is less than 10,000 Hz in the classical music composition for 1 to 3 minutes and accounts for the entire composition It is obtained by fermentation at a frequency of more than 99%; wherein the protection of the liver is to improve the antioxidant capacity of a hepatocyte or a DNA repair ability. The use as described in item 7 of the scope of patent application, wherein the DNA repairing ability is to improve the repairing ability of a DNA structural damage or a DNA double-strand break. The use as described in item 7 of the scope of patent application, wherein improving the antioxidant capacity of the hepatocyte includes increasing the gene expression of superoxide dismutase 1 (SOD1) and superoxide dismutase 2 (SOD2). The use as described in item 8 of the scope of patent application, wherein improving the repair ability of the DNA structural damage includes increasing the excision repair cross-complementing gene 1 (ERCC1) and DNA damage recognition and repair factor (DNA damage recognition and repair factor (XPA). The use as described in item 8 of the patent application scope, wherein improving the repair capacity of the DNA double-strand break includes adding X-ray repair cross complementation 1 (XRCC1) and X-ray repair cross complementation 2 (XRCC2) gene expression. The microbial fermentation product as described in item 1 of the scope of the patent application, wherein the total volume of the Streptococcus thermophilus, the yeast and the acetic acid bacteria is 2 to 7% of the volume of the vegetable juice and berry juice mixture.