Use of tangerine peel ferment for fat reduction

Abstract

The present application provides a use of a citrus peel ferment for preparing a fat-reducing composition. The citrus peel ferment is prepared by adding a peel of a citrus fruit to a yeast culture and fermenting at 25°C to 35°C for 3 to 10 days; wherein the yeast culture is obtained by adding 0.01% to 0.5% w / v of Saccharomyces cerevisiae to a yeast culture solution and culturing at 25°C to 35°C for 3 to 5 days; wherein the yeast culture solution is an aqueous solution comprising 1% to 5% w / v of yeast protein hydrolysate and 10% to 15% (w / v) of glucose. Furthermore, the prepared citrus peel ferment can assist obese people in losing weight through various mechanisms, such as reducing the fat content of fat cells, promoting the decomposition of triglycerides by fat cells, and increasing the expression of genes related to fat metabolism.

Description

[0001] This application is a divisional application filed by the applicant based on the parent application (application number: 201811621115.7, invention title: Orange peel fermentation product and its preparation method and application). Technical Field

[0002] This invention relates to the field of plant ferments, and more particularly to an orange peel ferment and its uses. Background Technology

[0003] A diet high in fat and sugar, coupled with insufficient physical activity, has led to obesity in many modern people. Obesity increases the risk of metabolic diseases such as diabetes, hyperlipidemia, hypertension, cardiovascular disease, and fatty liver, ultimately threatening health. Studies have also shown that obesity is a significant carcinogen. Furthermore, obese individuals are more prone to psychological problems and social difficulties. Therefore, many medical studies in recent years have focused on finding ways to curb obesity, attempting to promote physical and mental health in this way.

[0004] Methods to combat obesity include diet control, exercise, lifestyle changes, medication, and surgery. Except for severely obese patients who require surgery, most people rely on diet control and exercise to lose fat because modern people are often too busy to change their lifestyles and prefer natural therapies to unnecessary medications. However, strict dietary control requires a balanced diet and precise calorie intake, which is difficult to implement; improper exercise can cause injury. Furthermore, these two methods have limited effectiveness in fat loss because they do not directly target fat cells, especially visceral fat tissue.

[0005] In view of this, it is necessary to develop a composition that is easy for the general public to use and can effectively reduce fat accumulation in order to prevent obesity and reduce the likelihood of the aforementioned metabolic diseases and cancers. Summary of the Invention

[0006] Therefore, one object of the present invention is to provide a method for preparing orange peel ferment, comprising the following steps: (a) preparing a yeast culture containing brewer's yeast and a carbon source, and (b) adding orange peel to the yeast culture and fermenting it to obtain orange peel ferment.

[0007] In one embodiment of the present invention, the weight ratio of orange peel to yeast culture is 1:15 to 3:10.

[0008] In one embodiment of the present invention, the amount of brewing yeast is 0.01% to 0.5% of the weight of the yeast culture.

[0009] In one embodiment of the present invention, the carbon source is yeast protein hydrolysate and glucose.

[0010] In one embodiment of the present invention, the fermentation time is 3 to 10 days.

[0011] Another object of the present invention is to provide a fermented orange peel product prepared by the aforementioned preparation method.

[0012] Another object of the present invention is to provide the use of the aforementioned orange peel ferment for preparing a composition that reduces fat accumulation.

[0013] In one embodiment of the present invention, orange peel fermentation promotes the breakdown of triglycerides by adipocytes.

[0014] In one embodiment of the invention, orange peel fermentation promotes gene expression of low-density lipoprotein (LDL) receptor (LDLR), ATP-binding cassette transporter A1 (ABCA1), or combinations thereof in adipocytes.

[0015] In some embodiments, the use of an orange peel ferment for preparing a weight-loss composition is characterized in that the orange peel ferment is obtained by adding the peel of orange (Citrus reticulata) to a yeast culture and fermenting it at 25°C to 35°C for 3 to 10 days; wherein the yeast culture is obtained by adding 0.01% to 0.5% w / v Saccharomyces cerevisiae to a yeast culture broth and culturing it at 25°C to 35°C for 3 to 5 days; wherein the yeast culture broth is an aqueous solution containing 1% to 5% w / v yeast protein hydrolysate and 10% to 15% (w / v) glucose.

[0016] In some embodiments, the aforementioned weight-loss composition is prepared in unit dose form, wherein each unit dose contains 2 grams of orange peel ferment, for continuous application for at least 2 weeks to promote receptor weight loss.

[0017] This invention discloses that the aforementioned specific fermentation process can produce an orange peel ferment, which contains fat-reducing active ingredients not present in orange peel water extracts. This orange peel ferment can assist obese individuals in losing weight through multiple mechanisms, including reducing fat content in fat cells, promoting the breakdown of triglycerides in fat cells, and enhancing the expression of genes related to fat metabolism, resulting in a decrease in weight, waist circumference, body mass index (BMI), body fat percentage, and visceral fat index. Given that waist circumference, visceral fat content, and BMI are associated with the incidence of cardiovascular disease, the orange peel ferment has the potential to reduce the incidence of cardiovascular disease. Furthermore, consuming an orange peel fermented beverage containing this ferment can normalize circadian rhythm control to improve sleep disorders and inhibit the expression of cancer risk genes to reduce the risk of cancer. Therefore, this orange peel ferment can be used to prepare compositions that reduce fat accumulation, improve sleep quality, and reduce the risk of cancer. These compositions can be in the form of powder, granules, solution, colloid, or paste, and can be formulated as foods, beverages, pharmaceuticals, reagents, or nutritional supplements, administered orally to an individual.

[0018] The following will further illustrate the embodiments of the present invention with reference to the accompanying drawings. The examples listed below are intended to illustrate the inventive features and applications of the present invention, and are not intended to limit the scope of the present invention. Anyone skilled in the art can make some modifications and refinements without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention shall be determined by the appended claims. Attached Figure Description

[0019] Figure 1 This shows the relative fat content of adipocytes treated with orange peel ferment or orange peel water extract relative to the control group cells;

[0020] Figure 2 This shows the relative glycerol release from adipocytes treated with orange peel fermentation compared to control cells;

[0021] Figure 3 This shows the relative expression level of the LDLR gene in adipocytes treated with orange peel ferment or orange peel water extract compared to control cells;

[0022] Figure 4 This shows the relative expression level of the ABCA1 gene in adipocytes treated with orange peel ferment or orange peel water extract relative to the control group cells.

[0023] Figure 5 The study showed the weight changes of the subjects after they consumed the fermented orange peel beverage;

[0024] Figure 6 The study showed changes in waist circumference after the subjects consumed a fermented orange peel beverage.

[0025] Figure 7 The study showed changes in body mass index (BMI) after subjects consumed a fermented orange peel beverage.

[0026] Figure 8 The study showed changes in body fat percentage after subjects consumed a fermented orange peel beverage.

[0027] Figure 9 The study showed changes in visceral fat index in subjects after consuming a fermented orange peel beverage.

[0028] Figure 10 The study showed the relative expression level of the SIRT1 gene in peripheral blood mononuclear cells of subjects after 4 weeks of consuming the orange peel fermented beverage, compared to before consumption.

[0029] Figure 11 The study showed the sleep disorder scores of the subjects after consuming the orange peel fermented beverage for 4 weeks.

[0030] Figure 12A The study showed the relative expression levels of LAPTM4A and GPER genes in peripheral blood mononuclear cells of subjects after 4 weeks of consuming fermented orange peel beverage, compared to before consumption.

[0031] Figure 12B The study showed the relative expression levels of APP and SLPI genes in peripheral blood mononuclear cells of subjects after 4 weeks of consuming the orange peel fermented beverage, compared to before consumption.

[0032] Figure 12C The study showed the relative expression levels of HBB and HBA genes in peripheral blood mononuclear cells of subjects after 4 weeks of consuming fermented orange peel beverage, compared to before consumption.

[0033] Figure 12D The study showed the relative expression levels of BAX and SOCS3 genes in peripheral blood mononuclear cells of subjects after 4 weeks of consuming fermented orange peel beverage, compared to before consumption.

[0034] Figure 13 This shows the cancer risk multiplier after subjects consumed a fermented orange peel beverage for 4 weeks. Detailed Implementation

[0035] This invention provides a method for preparing an orange peel ferment, comprising the following steps: (a) preparing a yeast culture containing a brewer's yeast and a carbon source, and (b) adding an orange peel to the yeast culture and fermenting it to obtain an orange peel ferment. This invention also provides an orange peel ferment obtained by this method. The following examples reveal that the orange peel ferment can reduce the fat content of adipocytes, promote the breakdown of triglycerides in adipocytes, and enhance the expression of genes related to fat metabolism. Furthermore, consuming an orange peel fermented beverage containing this ferment not only helps obese individuals lose weight, but also normalizes circadian rhythm control to improve sleep disorders, and inhibits the expression of cancer risk genes to reduce the risk of cancer.

[0036] definition

[0037] The values ​​used in this article are approximate. All experimental data are expressed within 20%, the better range is within 10%, and the optimal range is within 5%.

[0038] Materials and Methods

[0039] Material

[0040] MEM-α medium (Minimum Essential Medium α medium), fetal bovine serum (FBS), penicillin / streptomycin, and phosphate-buffered saline (PBS) were purchased from Thermo Fisher Scientific. Oil Red O was purchased from Sigma-Aldrich. Formaldehyde and isopropanol were purchased from Echo Chemicals.

[0041] microorganism

[0042] Purchased Saccharomyces cerevisiae (BCRC 20271) from the Bioresource Collection and Research Center (BCRC) of the Food Industry Development Institute.

[0043] Cell culture

[0044] The following examples use adipocytes differentiated from the mouse mesenchymal cell line OP9 (ATCC CRL-2749), purchased from the American Type Culture Collection (ATCC). OP9 cells were differentiated at a ratio of 8 × 10⁻⁶. 4 Cells / well were seeded in 24-well culture dishes containing 500 μL of preadipocyte expansion medium (90% MEM-α, 20% FBS, 1% penicillin / streptomycin) and cultured at 37°C for 7 days. During cell culture, the medium was changed to adipocyte differentiation medium (90% MEM-α, 20% FBS, 1% penicillin / streptomycin) every 3 days. After 7 days, intracellular oil droplet formation was observed under a microscope (ZEISS; 400x magnification) to confirm complete differentiation into adipocytes.

[0045] In addition, human peripheral blood mononuclear cells (PBMCs) are isolated from the blood of the subjects.

[0046] Preparation of Oil Red O staining solution

[0047] To prepare a 3 mg / mL Oil Red O stock solution, completely dissolve the dye in 100% isopropanol. To obtain a usable Oil Red O dyeing solution, dilute the stock solution with deionized water to a concentration of 1.8 mg / mL immediately before use and filter through a 0.22 μm filter membrane.

[0048] Oil Red O staining

[0049] The lipid content in cells was determined by Oil Red O staining. Before staining, cells were washed with PBS and then fixed with 10% formaldehyde for 30 minutes. After washing once with PBS, the fixed cells were rinsed with 60% isopropanol for 1 minute. Subsequently, cells were stained with Oil Red O for 1 hour, the stain was removed, and then destained with 60% isopropanol for 5 seconds. After staining, cells were washed with PBS and observed under a microscope (ZEISS Axio Vert.A1), or the intracellular stain was dissolved in 100% isopropanol for 10 minutes for quantification. For quantification, 100 μL of the aforementioned stain-isopropanol solution was transferred into a 96-well plate, and the absorbance at 510 nm was measured using an ELISA (enzyme-linked immunosorbent assay) reader (BioTek). The quantitative results were statistically analyzed using the Student's t-test in Excel software.

[0050] Gene expression analysis

[0051] The expression levels of lipid metabolism-related genes in cells were determined using quantitative polymerase chain reaction (qPCR). The procedure is briefly described below. Following the manufacturer's instructions, RNA was isolated from cells using an RNA extraction kit (Geneaid). 2000 ng of RNA was reverse transcribed into cDNA at 37°C using SuperScript® III Reverse Transcriptase (Invitrogen). Subsequently, using the primer pair of the target gene and the β-actin gene ACTB (as an internal control) (Table 1), qPCR was performed on the aforementioned cDNA using a qPCR kit (KAPA CYBR FAST qPCR Kit (2X); KAPA Biosystems) in a StepOne Plus Real-Time PCR system (Applied Biosystems) to obtain melting curves.

[0052] Table 1

[0053]

[0054] Finally, using 2 -ΔΔCT The method measures the relative expression level of the target gene. This method uses the circulating threshold (C) of the ACTB gene. T The cycle threshold of the reference gene used as an internal control is used to calculate the relative fold change according to the following formula:

[0055] ΔC T = C of the target gene in the experimental or control group T C (internal control) T

[0056] ΔΔC T = ΔC of the experimental group T ΔC of the control group T

[0057] Multiple change = 2 ‐ ΔΔCt平均值

[0058] The expression levels of cancer risk genes in cells were determined using the methods described above. Table 2 shows the primer pairs of cancer risk target genes and the acidic ribosomal protein P0 (60S acidic ribosomal protein P0) gene RPLP0, which served as an internal control.

[0059] Table 2

[0060]

[0061]

[0062] The expression levels of sleep-related genes in cells were determined using the methods described above. Table 3 shows the primer pairs of sleep target genes and the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene, which served as an internal control.

[0063] Table 3

[0064]

[0065] Statistical analysis was performed using the STDEV function in Excel to calculate the standard deviation of the relative expression of each gene, and the statistical difference was calculated using the one-tailed Student's T-test (TTEST).

[0066] Example 1

[0067] Preparation of orange peel fermentation products

[0068] First, wash, dry, and chop the orange (Citrus reticulata) peel (approximately 0.5 to 1 cm on each side). Separately, prepare a yeast culture medium containing 1% to 5% (w / v) yeast peptone and 10% to 15% (w / v) glucose. This yeast culture medium can be heated at 50°C to 100°C for 0.5 to 2 hours, then cooled to room temperature. Add 0.01% to 0.5% (w / v) Saccharomyces cerevisiae (BCRC 20271) to this yeast culture medium and incubate statically at 25°C to 35°C for 3 to 5 days to obtain a yeast culture. Subsequently, the aforementioned orange peel and the yeast culture are mixed in a weight ratio of 1:15 to 3:10, and then allowed to ferment at 25°C to 35°C for 3 to 10 days to obtain an orange peel ferment.

[0069] The orange peel ferment can be concentrated under reduced pressure at 45°C to 70°C to obtain a concentrated orange peel ferment. This concentrated orange peel ferment can be further filtered through a 200-400 mesh filter to remove residual solids. The filtered concentrated orange peel ferment can then be selectively supplemented with 1-3% (w / w) citric acid and 40-70% (w / w) isomaltooligosaccharide, and sterilized to produce a ready-to-drink orange peel fermented beverage.

[0070] Example 2

[0071] The fat-reducing effects of fermented orange peel

[0072] To investigate the fat-reducing activity of the orange peel ferment of the present invention, an orange peel ferment was prepared according to the method described in Example 1 for a fat-reducing efficacy test. As a comparison, an unfermented orange peel aqueous extract was prepared by mixing orange peel with the yeast culture medium described in Example 1 at a weight ratio of 1:5 to 1:15 and extracting at 50°C to 100°C for 0.5 to 2 hours. During the experiment, 0.5% (w / w) of the orange peel ferment (experimental group 1) or 0.5% (w / w) of the orange peel aqueous extract (experimental group 2) was added to 24-well plates containing adipocytes differentiated from OP9 cells and cultured at 37°C for 7 to 10 days. The adipocyte differentiation medium containing 0.5% (w / w) of the orange peel ferment or 0.5% (w / w) of the orange peel aqueous extract was replaced every 3 days. Subsequently, the medium was removed, and the cells were washed with PBS solution for Oil Red O staining and determination of the relative fat content of each group of cells. The relative fat content is the ratio (expressed as a percentage) of the fat content of the experimental group cells to that of the control group cells. The control group cells were treated in a similar manner with adipocyte differentiation medium that did not contain the test sample.

[0073] Figure 1 The figure shows the relative fat content of adipocytes treated with orange peel ferment or orange peel aqueous extract compared to the control group. According to this figure, treatment with orange peel ferment significantly reduced the fat content of adipocytes compared to the control group, but orange peel aqueous extract did not have this effect. This result indicates that the orange peel fermentation process of the present invention may produce fat-reducing active ingredients not present in orange peel aqueous extract.

[0074] Example 3

[0075] Fat-decomposing effects of orange peel fermentation

[0076] To test the lipolysis effect of the orange peel ferment of this invention, adipocytes were treated with the orange peel ferment and the lipolysis effect was evaluated using a lipolysis assay. The lipolysis assay was performed using a glycerol cell-based assay kit (Caymen), which can quantify the glycerol released extracellularly due to the breakdown of intracellular triglycerides. In short, 0.25% (w / w) orange peel ferment (experimental group) was added to 24-well plates containing adipocytes differentiated from OP9 cells and cultured at 37°C for 7 to 10 days, during which the adipocyte differentiation medium containing 0.25% (w / w) orange peel ferment was replaced every 3 days. Subsequently, 25 μL / well of cell culture supernatant was transferred from the 24-well plate to a 96-well plate, and then the cell culture supernatant was mixed with 100 μL / well of free glycerol assay kit. After reacting the resulting mixture at room temperature for 15 minutes, the absorbance at 540 nm was measured using an ELISA reader (BioTek) to determine the relative glycerol release. This relative glycerol release is the ratio of glycerol release from the experimental group cells to that from the control group cells (expressed as a percentage). The control group cells were treated in a similar manner with adipocyte differentiation medium without orange peel fermentation.

[0077] Figure 2 The figure shows the relative glycerol release from adipocytes treated with orange peel fermentation compared to the control group. According to this figure, treatment with orange peel fermentation significantly increases glycerol release compared to the control group, indicating that the orange peel fermentation of this invention can promote the breakdown of triglycerides within adipocytes.

[0078] Example 4

[0079] Orange peel fermentation promotes the expression of genes related to fat metabolism.

[0080] To investigate the effects of orange peel fermentation on lipid metabolism, qPCR was used to determine the changes in the expression of lipid metabolism-related genes, including adenosine triphosphate (ATP)-binding cassette transporter A1 (ABCA1), scavenger receptor class B member 1 (SRB1, encoded by the SCARB1 gene), low-density lipoprotein receptor (LDLR), cholesterol ester transfer protein (CETP), and apolipoprotein A1 (APOA1), in adipocytes differentiated from OP9 cells after treatment with orange peel fermentation. In short, the effects of orange peel fermentation on lipid metabolism were investigated by qPCR. 5Cells were seeded per well in a 6-well dish containing 2 mL of adipocyte differentiation medium. After incubation at 37°C for 24 hours, the medium was removed, and the cells were washed with PBS. Subsequently, cells were treated with 1 mL of adipocyte differentiation medium containing 1 mg / mL orange peel ferment (Experimental Group 1) or 1 mg / mL orange peel aqueous extract (Experimental Group 2), or treated with adipocyte differentiation medium alone as a control group. All three groups of cells were cultured at 37°C for 48 hours before being used for qPCR analysis.

[0081] Figure 3 This shows the relative expression level of the LDLR gene in adipocytes treated with orange peel ferment or orange peel water extract compared to control cells; Figure 4 The figures show the relative expression levels of the ABCA1 gene in adipocytes treated with orange peel fermentation product or orange peel water extract compared to the control group cells. *** in each figure indicates p < 0.001. Based on... Figure 3-4 Compared to the control group, treatment with either orange peel ferment or orange peel water extract significantly increased the relative expression level of the LDLR gene, with orange peel ferment treatment increasing LDLR gene expression by approximately 2.2 times. However, only orange peel ferment treatment significantly increased ABCA1 gene expression by approximately 5.7 times. This result indicates that the orange peel ferment of the present invention not only promotes the metabolism of low-density lipoprotein cholesterol by hepatocytes but also promotes the synthesis of high-density lipoprotein (HDL), thus helping to reduce cholesterol accumulation on blood vessel walls and maintain cardiovascular health.

[0082] Example 5

[0083] Fermented orange peel promotes weight loss

[0084] To test whether the orange peel ferment of the present invention has a weight loss effect, eight obese subjects (body fat percentage greater than 27% or BMI greater than 24) drank the orange peel fermented beverage (containing 2 g of orange peel ferment) prepared according to the method described in Example 1 daily for 4 weeks. The weight loss indicators of these subjects were measured by body composition analyzer (TANITA BC-601FS) before drinking (week 0), after drinking for 2 weeks, and after drinking for 4 weeks, including weight, waist circumference, body mass index (BMI), body fat percentage, and visceral fat index.

[0085] Figure 5 , 6 Tables 7, 8, and 9 show the average changes in weight, waist circumference, BMI, body fat percentage, and visceral fat index of the aforementioned subjects after consuming the fermented orange peel beverage; * and ** in the figures indicate p < 0.05 and p < 0.01 compared to week 0, respectively. Based on... Figure 5-9Compared to before consumption, drinking the fermented orange peel beverage for four consecutive weeks resulted in a weight loss of approximately 1.1 kg, a waist circumference reduction of approximately 2.6 cm, a BMI reduction of approximately 0.4, a body fat percentage reduction of approximately 0.6%, and a visceral fat index reduction of approximately 0.5. These results confirm that long-term use of the fermented orange peel product of this invention can improve the weight loss index in obese individuals.

[0086] Example 6

[0087] Orange peel fermentation improves sleep disorders

[0088] To test whether the fermented orange peel of this invention has additional effects in improving sleep disturbances, six subjects with sleep disorders drank a fermented orange peel beverage (containing 2 g of fermented orange peel) prepared according to the method described in Example 1 daily for four weeks. Peripheral blood mononuclear cells were collected from these subjects before and after four weeks of drinking to analyze changes in the expression levels of sleep-related genes (such as the SIRT1 gene) in these cells using qPCR. In addition, these subjects also underwent a sleep questionnaire survey to assess changes in the degree of sleep disturbance before and after drinking the beverage in a self-controlled manner. The degree of sleep disturbance was judged based on a sleep disturbance score, which was the sum of the scores given by the subjects according to the assessment items listed in Table 4 below.

[0089] Table 4

[0090]

[0091] Figure 10 This figure shows the relative expression level of the SIRT1 gene in peripheral blood mononuclear cells of the aforementioned subjects after consuming the fermented orange peel beverage, compared to before consumption. According to this figure, consuming the fermented orange peel beverage for four consecutive weeks increased SIRT1 gene expression by approximately eight times compared to before consumption (week 0). Given that previous studies have shown that the SIRT1 protein can regulate the expression of diurnal regulation genes to establish circadian rhythms, and that its expression level decreases with age, these experimental results indicate that long-term use of the fermented orange peel product of this invention is beneficial for normal circadian rhythm control and may even help older adults maintain a normal biological clock.

[0092] Figure 11 The figure shows the change in the average sleep disorder score of the aforementioned subjects after consuming the fermented orange peel beverage. According to this figure, compared to before consumption (week 0), continuous consumption of the fermented orange peel beverage for 4 weeks significantly reduced the subjects' sleep disorders by approximately 26%. This result indicates that long-term use of the fermented orange peel product of this invention can improve sleep disorders.

[0093] Example 7

[0094] Orange peel fermentation reduces cancer risk

[0095] To test whether the orange peel ferment of the present invention reduces the risk of cancer, six subjects drank an orange peel fermented beverage (containing 2 g of orange peel ferment) prepared according to the method described in Example 1 daily for 4 weeks. Peripheral blood mononuclear cells of these subjects were collected before drinking and after 4 weeks of drinking in order to analyze the changes in the expression levels of cancer risk genes in the cells using qPCR. Cancer risk genes are classified into four groups, including (1) genes that promote cell growth and evade apoptosis, including genes for lysosome-associated membrane protein 4α (LAPTM4A) and G protein-coupled receptor (GPER); (2) genes that promote cell migration and invasion, including genes for amyloid precursor protein (APP) and secretory leukocyte protease inhibitor (SLPI); (3) genes that supply nutrients to the blood circulation, including genes for hemoglobin subunit beta (HBB) and hemoglobin subunit α (HBA); and (4) genes that evade the immune system, including genes for Bcl-2-associated X protein (BAX) and cytokine signaling pathway inhibitor 3 (SOCS3).

[0096] Figure 12A-12D This figure shows the relative expression levels of cancer risk genes in peripheral blood mononuclear cells of the aforementioned subjects after consuming the fermented orange peel beverage, compared to before consumption. *, **, and *** in the figure represent p < 0.05, p < 0.01, and p < 0.001, respectively. Based on... Figure 12A-12D Compared to before drinking (week 0), drinking fermented orange peel beverage for 4 weeks can generally inhibit the expression of the aforementioned four cancer risk gene groups, especially the expression of genes such as LAPTM4A, GPER, APP, SLPI, HBA, BAX, and SOCS3.

[0097] By comparing the relative expression levels of the aforementioned cancer risk genes with the TCI Gene Database (constructed from genomic data of sub-healthy individuals), the cancer risk multiplier of the subjects before and after consuming the orange peel fermented beverage was estimated. Figure 13 As shown in the figure, drinking fermented orange peel beverage for four consecutive weeks can reduce the cancer risk factor, that is, reduce the likelihood of developing cancer. The aforementioned results indicate that long-term use of the fermented orange peel product of this invention can reduce the risk of cancer.

[0098] In summary, the above experiments demonstrate that the orange peel fermentation process of the present invention can produce an orange peel ferment, which contains fat-reducing active ingredients not present in orange peel water extracts. This orange peel ferment can assist obese individuals in weight loss through multiple mechanisms, including reducing fat content in fat cells, promoting the breakdown of triglycerides in fat cells, and enhancing the expression of genes related to fat metabolism. Furthermore, consuming an orange peel fermented beverage containing this ferment can normalize circadian rhythm control to improve sleep disorders and inhibit the expression of cancer risk genes to reduce the risk of cancer. Therefore, this orange peel ferment can be used to prepare compositions that reduce fat accumulation, improve sleep quality, and reduce the risk of cancer. These compositions can be in the form of powder, granules, solution, colloid, or paste, and can be formulated as food, beverage, pharmaceutical, reagent, or nutritional supplement, administered orally to an individual.

Claims

1. Use of a citrus peel ferment for the preparation of a slimming composition, characterized in that, The orange peel ferment is prepared by adding the peel of orange (Citrus reticulata) to a yeast culture and fermenting it at 25°C to 35°C for 3 to 10 days. The yeast culture is obtained by adding 0.01% to 0.5% w / v Saccharomyces cerevisiae to a yeast culture medium and culturing it at 25°C to 35°C for 3 to 5 days. The yeast culture medium is an aqueous solution containing 1% to 5% w / v yeast protein hydrolysate and 10% to 15% (w / v) glucose.

2. Use according to claim 1, characterized in that, The weight ratio of the pericarp to the yeast culture is 1:15 to 3:

10.

3. Use according to claim 1, characterized in that, The orange peel ferment has the ability to promote the breakdown of triglycerides by adipocytes.

4. The use according to claim 1, characterized in that, The orange peel ferment has the ability to promote gene expression of low-density lipoprotein receptor, adenosine triphosphate binding cassette transporter A1, or a combination thereof in adipocytes.

5. The use according to claim 1, characterized in that, The weight loss composition is prepared in unit dose form, wherein each unit dose contains 2 grams of the orange peel ferment, for continuous application for at least 2 weeks to promote recipient weight loss.

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