TWM590969U - Particle structure of rutaceae plant fermentation broth with biomimetic stroma system - Google Patents

Abstract

This creation provides a microstructure of the fermentation broth of a Rutaceae plant with a biomimetic interstitial system, including a protective outer layer, which is transformed and divided into small molecular polysaccharides by polysaccharide molecules contained in a special fermentation of a mushroom and an algae Molecule, which becomes a coating material, and an inner layer containing functional ingredients, which is a substantially circular shape containing functional substances in the fermentation broth of rutaceae plants (such as citrus). The diameter of the outer layer is between 10 ^-4 ~10 ^-3 mm The particle size of the inner layer is between 10 ^-5 ~10 ^-4 mm, and the inner layer is coated in the outer layer. This double-layer structure can retain the fat-loss and weight-loss, anti-aging and anti-wrinkle effects of the fermentation broth of rutaceae plants (such as citrus), and supplement the glycosides lost by the functional substances during the fermentation process, improve the utilization rate of the human body, and effectively improve the efficiency Even if the output has not been improved, the function of the active ingredient can also achieve the same effect, even better than the efficiency of the natural structure.

Description

Microstructure of fermentation broth of Rutaceae plant with biomimetic interstitial system

This creation provides a microstructure of the fermentation broth of the Rutaceae plant that mimics the biological interstitial system. It contains a protective outer layer, which is transformed and divided into small molecular polysaccharide molecules by the polysaccharide molecules contained in a special fermentation solution of a mushroom and an algae. It becomes a coating material, and an inner layer contains functional substances in the fermentation broth of rutaceae plants (such as citrus). The diameter of the outer layer is between 10 ^-4 ~10 ^-3 mm. Between 10 ^-5 ~10 ^-4 mm, the protective outer layer covers the inner layer. This double-layer structure can retain the fat-loss and weight-loss, anti-aging and anti-wrinkle effects of the fermentation broth of rutaceae plants (such as citrus), and supplement the glycosides lost by the functional substances during the fermentation process, improve the utilization rate of the human body, and effectively improve the efficiency Even if the output has not been improved, the function of the active ingredient can also achieve the same effect, even better than the efficiency of the natural structure.

Citrus is a general term for citrus fruits in the Rutaceae family. The main types are tangerines, mandarins, sweet oranges, limes, pomelo, grapefruit, lemon, lime, citrus, bergamot, and kumquat. The citrus fruits are The world's largest fruit production, the current annual output has exceeded 100 million tons; citrus is rich in potassium, B vitamins, vitamin C and antioxidant ingredients, anti-cancer ingredients, anti-allergic ingredients, but also rich in flavonoids, Polyphenols, carotenoids and other compound groups have high nutritional value and dietary health care. In addition, citrus has the functions of fat loss, weight loss, antioxidant and anti-aging, but the current technology is still water extraction or organic solvents. Extraction is the main method of extraction. Because the effective ingredients are in the form of natural compounds, they are easily affected by the season and weather of the raw materials.

Current research indicates that the natural compounds (such as flavonoids) in citrus are not highly utilized in the human body, and the glycosides are easily removed during the extraction process, which reduces the absorption of the human body, and the types of citrus extracted are also limited. Obtaining higher levels of functional substances in accordance with different citrus types can increase the original efficiency in order to increase the bioavailability of citrus, and the utilization energy is not limited to different citrus types, which is an urgent problem to be solved.

According to the literature Zhang, Y. et al, (2010) Comparative Analysis of Contents of Flavonoid Compounds in Various Species of Citrus Peel and Their Dynamic Changes in Quzhou Ponkan Peel during Postharvest Storage, the study adopted naringin in different varieties of citrus peel , Hesperidin, Neohesperidin, Chuancetin, Hesperetin, Orange Flavone were analyzed and determined to study the distribution and differences of the main flavonoid compounds in different varieties of citrus peel, as well as the content of flavonoid compounds in the citrus peel after harvest The results showed that the composition and content of flavonoids in different varieties of citrus fruits were significantly different. Among the five citrus varieties, the total content of six flavonoids in the peel of Changshan Huyou The highest, mainly naringin and neohesperidin, accounting for 98.4% of the total content of the 6 types of flavonoids; Mengchuan Wenzhou mandarin oranges and Hamlin sweet orange peels are mainly hesperidin, accounting for 6 species The total content of flavonoids was 96.9% and 94.2%, while the peels of Quzhou Ponkan and Banana were mainly hesperidin and ligustrazine, accounting for 86.5% and 91.2% of the six flavonoid compounds, respectively. The content distribution of 6 flavonoids in Ponkan and Banana was consistent, all of which were hesperidin> sialonin> hesperetin> hesperidin> naringin> neohesperidin. Therefore, the content of flavonoids in different citrus species is different, and the flavonoids are mainly distributed in the peel.

Therefore, the development of a rutinaceae plant fermentation broth composition suitable for general public use, improving human utilization and improving the effectiveness of efficacy ingredients, and having the functions of reducing fat and weight, and anti-aging and anti-wrinkle are the most important problems to be solved at this stage.

Unless otherwise defined, all technical terms used herein have the same meaning as those of ordinary skill in the art to which this creation belongs. Unless the context clearly indicates otherwise, the singular forms "a", "an", etc. used in this specification and the appended claims include plural references. Unless otherwise stated, any reference to "or" in this document is intended to cover "and/or".

The following will provide some explanations of this creation. These explanations present the selected concepts of this creation in a simplified form as an introduction to the more detailed description presented below.

The present invention provides a microstructure of fermented broth of Rutaceae plant, which comprises, in order from the center to the outside: an inner layer, which contains a fermented broth of Rutaceae plant, and a protective outer layer, which contains a mushroom and a The polysaccharide molecules contained in the algae after special fermentation are transformed and divided into small molecular polysaccharide molecules. Among them, the microstructure of the fermentation broth of the Rutaceae plant has the effect of reducing fat and weight or anti-aging and anti-wrinkle.

In one embodiment of the present invention, the diameter of the protective outer layer with fat-reducing, weight-reducing, anti-aging and anti-wrinkle effect in the microstructure of the Rutaceae plant fermentation broth is between 10 ^-4 ~10 ^-3 mm; the inner layer The particle size is between 10 ^-5 ~10 ^-4 mm.

In one embodiment of the present invention, the rutaceae plant contained in the inner layer of the fermented broth of the rutaceae plant with the effect of reducing fat, weight loss, anti-aging and wrinkle is selected from orange, lemon, lime, kumquat and their Groups of mixtures.

In an embodiment of the present invention, the protective outer layer wherein the mushrooms are selected from the group consisting of shiitake mushrooms, black fungus, white fungus and mixtures thereof.

In an embodiment of the present invention, the protective outer layer wherein the algae is selected from the group consisting of cyanidia, sea fungus, kelp and mixtures thereof.

In one embodiment of the present invention, the protective outer layer structure can retain the fat-loss and weight-loss, anti-aging and anti-wrinkle effects of the fermentation broth of Rutaceae plants, and supplement the glycosides lost by the functional substances during the fermentation process to improve the utilization rate of the human body , And effectively improve the efficiency, even if the output has not been improved, the function of the active ingredient can also achieve the same effect, even better than the natural composition The effectiveness of the structure.

In one embodiment of the present invention, the microstructure of the fermentation broth of the new rutaceae plant can be manufactured in the form of powder packets, drinks, capsules or lozenges for users to choose according to their needs.

In one embodiment of the present invention, the method for preparing the microstructure of the fermentation broth of the Rutaceae plant includes:

(1) Fermentation step of lactic acid bacteria: the independent fermenter is used to make the whole rutaceae plant mass at least 50-80%, add one lactic acid bacteria and one sugar 0.2-20%, carbon dioxide is controlled below 1000ppm, temperature setting It is 25-28℃, the pH value is adjusted to between 5-7, fermented for 30-60 days, obtain a lactic acid fermented fermented liquid of a Rutaceae plant.

(2) Yeast fermentation step: add the yeast lactic acid fermentation solution of Rutaceae plant to a yeast and a sugar 0.2-20%, and the carbon dioxide is controlled below 1000ppm, the temperature is set to 25-28 ℃, the pH value is adjusted Between 5-7, 30-60 days of fermentation, obtain a yeast fermentation solution of Rutaceae plants.

(3) Mixed fermentation step: add a mushroom or algae fermentation broth, the fermentation broth process is the same as the first two steps of the fermentation broth of Rutaceae plants, additional sugars (such as: fructose, glucose, lactose, xylose) ) 0.2-20%, carbon dioxide is controlled below 1000ppm, the temperature is set at 25-28°C, and the fermentation days are 30-60 days. The rutinaceae plant fermentation broth composition is used for the purpose of preparing fat loss, weight loss, anti-aging and anti-wrinkle.

In the preparation method described in the novel microbial structure of the Rutaceae plant fermentation broth, the mushrooms are selected from the group consisting of mushrooms, black fungus, white fungus, or a mixture thereof.

In the preparation method described in the novel microbial structure of the Rutaceae plant fermentation broth, the algae are selected from the group consisting of cyanobacteria, sea fungus, kelp or mixtures thereof.

In the preparation method described in the novel microbial structure of Rutaceae plant fermentation broth, the carbohydrate is selected from the group consisting of fructose, glucose, lactose or xylose.

In the preparation method described in the novel microbial structure of the Rutaceae plant fermentation broth, the lactic acid bacteria are selected from the group consisting of Lactobacillus plantarum, Lactobacillus delbrueckii, Lactococcus lactis, Lactococcus acidophillus or Bifidobacterium bifidum.

In the preparation method described in the novel microbial structure of Rutaceae plant fermentation broth, the mixing ratio of the lactic acid bacteria is Lactobacillus plantarum concentration 0.5%~10%, Lactobacillus delbrueckii concentration 0.5%-10%, and Lactococcus lactis concentration 0.5%-10 %, Lactococcus acidophillus concentration is 0.5%~10% and Bifidobacterium bifidum concentration is 0.5%~10%.

In the preparation method described in the novel microbial structure of Rutaceae plant fermentation broth, the yeast is selected from the group consisting of Saccharomycopsis fibufigera, Pichia membrame faciens, Schizosaccharomyes pombe or Saccharomyces cerevisiae.

In the preparation method described in the novel microbial structure of Rutaceae plant fermentation broth, the mixing ratio of the yeast is Saccharomycopsis fibufigera concentration is 0.5%~10%, Pichia membrame faciens concentration is 0.5%~10%, Schizosaccharomyes pombe concentration is 0.5%~10% and the concentration of Saccharomyces cerevisiae is 0.5%~10%. In the preferred preparation method described in the microstructure of the fermentation broth of the new rutaceae plant, the lactic acid bacteria Lactococcus acidophillus is added to a rutaceae plant for 30 days, then the yeast Saccharomycopsisfibufigera is added for 30 days, and then a mushroom or an algae The fermentation broth is mixed and fermented for 30 days to form a broth plant fermentation broth composition, also known as a rutaceae composite crystal.

10‧‧‧ Rutaceae

11‧‧‧ Rutaceae plant lactic acid bacteria fermentation step

12‧‧‧ Rutaceae plant yeast fermentation steps

20‧‧‧Mushrooms and algae

21‧‧‧ Steps for fermentation of mushrooms and algae lactic acid bacteria

22‧‧‧ Fermentation steps for mushrooms and algae yeast

30‧‧‧ Mixed fermentation

40‧‧‧Particulate structure of Rutaceae plant fermentation broth (Rutaceae composite crystal)

401‧‧‧Protect the outer layer

402‧‧‧Inner layer

50‧‧‧Broken wall extraction

60‧‧‧Structural modification

70‧‧‧ Crystal construction

The first figure is a schematic diagram of the method for preparing the microstructure of the fermentation broth of the Rutaceae plant.

The second figure shows an anatomical view of one embodiment of the microstructure of the fermentation broth of the Rutaceae plant (that is, a composite crystal of Rutaceae).

The third figure shows the UCP-1 brown adipose tissue protein conversion test of the microstructure of the Rutaceae plant fermentation broth (that is, the Rutaceae composite crystal). The experiments are divided into six groups: (1) 0.25% Rutaceae water extract, (2) 0.125 % Rutaceae water extract, (3) 0.25% Rutaceae fermentation broth, (4) 0.125% Rutaceae fermentation broth, (5) 0.25% Rutaceae composite crystal and (6) 0.125% Rutaceae composite crystal, the value is expressed as Mean ± standard deviation (n=3), 3T3-L1 preadipocytes were co-cultured with 6 groups of samples and transformed into brown adipocytes (the UCP-1 content increased) can increase the fat burning effect: the results show that Rutaceae compound The crystals show a good conversion effect. Compared with the rut extract and rut fermentation broth, the conversion rate can reach 3 times within 8 hours with 0.25% rutaceae composite crystals. And the rutinaceae composite crystals can be continuously transformed for up to 8 hours, while the rut extract and rut fermentation liquid can only hold It lasts for 6 hours, so after the fermentation process, the efficacy of rutaceae can be improved, and the browning ability of fat cells can be improved.

The fourth figure shows the pre-adipocyte inhibitory capacity test of the microstructure of the Rutaceae plant fermentation broth (that is, the Rutaceae composite crystal). The toxicity test after NIH-3T3 fibroblasts were treated with 13 groups of samples for 24 hours, the results showed 24 After incubation for hours, none of the extracts of rut perfume, rut fermentation broth, and rutaceae composite crystals showed any toxicity. The survival rate of fibroblasts exceeds 85%, with no adverse effects. Values are expressed as mean ± standard deviation, and cell viability exceeding 80% indicates no cytotoxicity.

The fifth figure shows the pre-adipocyte inhibitory capacity test of the microstructure of the Rutaceae plant fermentation broth (that is, the Rutaceae composite crystal). The toxicity test after NIH-3T3 fibroblasts were treated with 13 groups of samples for 48 hours, the results showed 48 After incubation for hours, none of the extracts of rut perfume, rut fermentation broth, and rutaceae composite crystals showed any toxicity. The survival rate of fibroblasts exceeds 85%, with no adverse effects. Values are expressed as mean ± standard deviation, and cell viability exceeding 80% indicates no cytotoxicity.

The sixth figure shows the preadipocyte inhibitory ability test of the microstructure of the Rutaceae plant fermentation broth (that is, the Rutaceae composite crystal), and the results show that the 3T3-L1 preadipocytes and the 13 groups of samples were inhibited by the adipocytes after 24 hours, This inhibitory effect: Rutaceae composite crystals> rut perfume extraction> rut fermentation liquid, the general fermentation method can not improve the efficacy components in citrus; Rutaceae composite crystals show significant ability to inhibit pre-adipocytes, 24-hour survival rate 65%, effectively reduce the formation of fat cells. Values are expressed as mean ± standard deviation. In the figure, ap<0.01 indicates a significant difference compared with the control group; bp<0.01 indicates a significant difference compared with the Rutaceae water extract group; cp<0.01 indicates Compared with Rutaceae fermentation broth, there are significant differences.

The seventh figure shows the preadipocyte inhibitory ability test of the microstructure of the Rutaceae plant fermentation broth (that is, the Rutaceae composite crystal). The results show that the 3T3-L1 preadipocytes and the 13 groups of samples were inhibited by the adipocytes after 48 hours, This inhibitory effect: Rutaceae composite crystals> rut perfume extraction> rut fermentation liquid, which cannot improve the efficiency of citrus components in general fermentation; Rutaceae composite crystals show significant ability to inhibit pre-adipocytes, with a 48-hour survival rate of 50 %, effectively reduce the generation of fat cells. Values are expressed as mean ± standard deviation. In the figure, ap<0.01 indicates a significant difference compared with the control group; bp<0.01 indicates a significant difference compared with the Rutaceae water extract group; cp<0.01 indicates Compared with Rutaceae fermentation broth, there are significant differences.

The eighth figure shows the antioxidant capacity test of the microstructure of the Rutaceae plant fermentation broth (that is, the Rutaceae composite crystal). The experiment is divided into six groups of samples, namely: (1) 0.125% Rutaceae water extract, (2) 0.25% Rutaceae water extract, (3) 0.125% rutaceae fermentation broth, (4) 0.25% rutaceae fermentation broth, (5) 0.125% rutaceae composite crystal and (6) 0.25% rutaceae composite crystal, with common free radicals : DPPH is used as an indicator. In vitro tests were carried out. Six groups of samples with different concentrations were added to test whether the samples have the ability to scavenge free radicals. The results show that the ability to scavenge DPPH free radicals: rut Branch compound crystal> Rut perfume extract> Rut fermentation liquid, wherein the rut branch compound crystal achieves 63% removal effect. The values are expressed as mean ± standard deviation. In the figure, a.p<0.05 indicates a significant difference compared with the Rutaceae water extract group; b.p<0.05 indicates a significant difference compared to the Rutaceae fermentation broth.

The ninth figure shows the antioxidant capacity test of the microstructure of Rutaceae plant fermentation broth (that is, Rutaceae composite crystal). The experiment is divided into six groups of samples, respectively: (1) 0.125% Rutaceae water extract, (2) 0.25% Rutaceae water extract, (3) 0.125% rutaceae fermentation broth, (4) 0.25% rutaceae fermentation broth, (5) 0.125% rutaceae composite crystal and (6) 0.25% rutaceae composite crystal, with common free radicals : ABTS+ is used as an indicator to conduct an in vitro test. Six groups of samples with different concentrations are added to test whether the samples have the ability to scavenge free radicals. The results show that the ability to scavenge ABTS+ free radicals: Rutaceae composite crystals> rut perfume extract> rut fermentation broth, where The Rutaceae composite crystal achieves a 57% removal effect. The values are expressed as mean ± standard deviation. In the figure, a.p<0.05 indicates a significant difference compared with the Rutaceae water extract group; b.p<0.05 indicates a significant difference compared to the Rutaceae fermentation broth.

The tenth figure shows the antioxidant capacity test of the microstructure of Rutaceae plant fermentation broth (that is, Rutaceae composite crystal). The experiment is divided into six groups of samples, namely: (1) 0.125% Rutaceae water extract, (2) 0.25% Rutaceae water extract, (3) 0.125% rutaceae fermentation broth, (4) 0.25% rutaceae fermentation broth, (5) 0.125% rutaceae composite crystal and (6) 0.25% rutaceae composite crystal, with common free radicals : NBT is used as an indicator to conduct in vitro tests, adding 6 groups of samples with different concentrations respectively The test samples have the ability to scavenge free radicals, and the results show the ability to scavenge NBT free radicals: rutaceae composite crystals> rut perfume extract> rut fermentation liquid, in which the rut family compound crystals achieve 90% scavenging effect. The values are expressed as mean ± standard deviation. In the figure, a.p<0.05 indicates a significant difference compared with the Rutaceae water extract group; b.p<0.05 indicates a significant difference compared to the Rutaceae fermentation broth.

The eleventh figure shows the tyrosinase inhibitory activity test of the microstructure of Rutaceae plant fermentation broth (that is, the compound crystal of Rutaceae). The experiment is divided into eight groups of samples, and tyrosine is catalyzed by tyrosinase to produce dopa. The melanin precursor, the results show that the samples of each group are concentration-dependent and inhibit the tyrosinase activity test results: kojic acid> rutaceae composite crystal> rutaceae water extract> rutaceae fermentation broth; 76% inhibition rate, reduce the production of melanin precursors. Values are expressed as mean ± standard deviation. In the figure, a.p<0.01 indicates a significant difference compared with the Rutaceae water extract group; b.p<0.01 indicates a significant difference compared to the Rutaceae fermentation broth.

The twelfth figure shows the MMP-2 protein performance test of the microstructure of the fermentation broth of the Rutaceae plant (that is, the compound crystal of the Rutaceae). The experiment is divided into nine groups of samples. Pro MMPs are unactivated MMPs. If the concentration of Pro MMPs increases, it means Reduced the activation of MMPs and achieved the maintenance of collagen. The results of MMP-2 protein expression showed that the samples of each group were concentration-dependent; Pro MMP-2 protein concentration: Rutaceae composite crystal> Vitamin C> Rutaceae water extract> Rutaceae fermentation broth, in which the rutaceae composite crystals increase the concentration of Pro MMP-1 to avoid the activation of MMP-2 Collagen lysis. Values are expressed as mean ± standard deviation. In the figure, a.p<0.01 indicates a significant difference compared with the Rutaceae water extract group; b.p<0.01 indicates a significant difference compared to the Rutaceae fermentation broth.

The thirteenth figure shows the experiment of obese mice with microstructure of Rutaceae plant fermentation broth (ie, Rutaceae composite crystal 40). The experiment is divided into 2 groups of samples, namely: (1) Control group (without giving Rutaceae composite crystal ) And (2) Rutaceae compound crystals, feeding mice with high fat feed to induce obesity, and at the same time giving rutaceae compound crystals to monitor the weight change rate of mice, the test results of obese mice showed little feeding with high fat feed In mice, the weight growth rate increased with time, and the weight gain rate reached 39% on the 19th day; mice with rutinaceous compound crystals can reduce the rate of change in body weight and reduce the weight gain rate by 8%.

In order to provide a more complete and clear disclosure of the technical content used in this creation, the purpose of the creation and the effect achieved, please refer to the detailed description below, and please refer to the drawings and figures.

Example 1: Method for preparing microstructure of fermented broth of Rutaceae plant

Through microbial fermentation technology, the biological characteristics are used to consume the structural components of the active ingredients (such as flavonoids), and then the small molecule mushroom or algae polysaccharide peptide is added to solve the problem of reducing the utilization rate due to extraction. In the fermentation process: first extraction, later modification, and then construction, three-stage fermentation technology, The fermentation broth of the rutaceae plant with a crystal structure is produced, which has the functions of reducing fat and weight, and anti-aging and anti-wrinkle. The fermentation process of the microstructure 40 of the rutaceae plant fermentation broth particulate structure is shown in the first figure, including:

1. Lactic acid bacteria fermentation step 11:

Using microbial characteristics to consume the active components of rutaceae plants (such as flavonoids) structural glycosides, breaking the wall to extract 50 rutaceae plants.

At the time of preparation, the quality of the complete rutaceae plants 10 (such as orange, lemon, lime, kumquat) accounted for at least 50~80%, and the lactic acid bacteria 1~5 strains (Lactobacillus plantarum, Lactobacillus delbrueckii, Lactococcuslactis, Lactococcusacidophillus, Bifidobacterium bifidum), sugars (such as fructose, glucose, lactose, xylose) 0.2~20%, carbon dioxide is controlled below 1000ppm, the temperature is set at 25~28℃, and the pH value is adjusted to between 5~7 , Fermentation days 30~60 days.

2. Yeast fermentation step 12:

The rudimentary plants 10 (such as orange, lemon, lime, kumquat) that are initially decomposed are small molecules, using digestive enzymes or hydrolytic enzymes, and the structural modification of the rutaceous plant structure is carried out at the same time, and the yeast is fermented During the process, the excess monosaccharide molecules on the structure of nutrients are decomposed and used. After the structure modification, the nutrients can have higher adhesion characteristics, which is conducive to subsequent advanced modification (such as glycosylation)60.

At the time of preparation, the quality of the complete rutaceae plants (such as orange, lemon, lime, kumquat) should be at least 50~80%, and 1~4 yeasts should be added. (Saccharomycopsisfibufigera, Pichia membramefaciens, Schizosaccharomyespombe, Saccharomyces cerevisiae), sugars (such as fructose, glucose, lactose, xylose, brown sugar) 0.2~20%, carbon dioxide is controlled below 1000ppm, temperature is set at 25~28℃, pH value is adjusted Between 5 and 7, the fermentation days are 30 to 60 days.

3. Step 30 of the mixed fermentation stage:

Add mushroom 20 (such as shiitake mushrooms, black fungus, white fungus) or algae 20 (such as blue algae, sea fungus, kelp) fermentation broth, lactic acid bacteria fermentation step 21, lactic acid bacteria fermentation step 22 and the Rutaceae plant in the fermentation broth process The first two stages of the fermentation broth are the same as 11, 12. Add extra sugars (such as fructose, glucose, lactose, xylose) 0.2-20%, carbon dioxide is controlled below 1000ppm, the temperature is set at 25-28 ℃, the fermentation days 30- 60 days. That is, in the fermentation process: first extraction, then modification, and then construction, three-stage fermentation technology, to produce a microstructure 40 of a rutile plant fermentation broth with a crystal structure.

Example 2: Microstructure of Rutaceae plant fermentation broth 40

The novel rutaceae plant fermentation broth particle structure 40 includes, in order from the center to the outside, an inner layer 402 that contains a rutaceae plant fermentation broth; and a protective outer layer 401, where the protective outer layer includes a mushroom The polysaccharide molecules contained in special fermented yeasts and algae are transformed and divided into small molecular polysaccharide molecules. Among them, the microstructure of the fermentation broth of Rutaceae plant has the effect of reducing fat and weight or anti-aging and anti-wrinkle.

The novel microstructure 40 of the Rutaceae plant fermentation broth. The diameter of the protective outer layer 401 in the microstructure of the Rutaceae plant fermentation broth with the effect of reducing fat, weight, and anti-aging and wrinkle is between 10 ^-4 ~10 ^-3 mm ; The particle size of the inner layer 402 is between 10 ^-5 ~10 ^-4 mm.

The novel microstructure 40 of the Rutaceae plant fermentation broth, the rutaceae plant contained in the inner layer 402 of the Rutaceae plant fermentation broth with the functions of reducing fat and weight and having anti-aging and anti-wrinkle effects is selected from the group consisting of orange, lemon, lime and gold Orange and its mixture.

The novel microstructure 40 of the Rutaceae plant fermentation broth has a fat-reducing, weight-reducing, anti-aging and wrinkle-resistant Rutaceae plant fermentation broth protective outer layer 401. The mushrooms are selected from the group consisting of shiitake mushrooms, black fungus, white fungus and mixtures thereof Group.

In the novel rutaceae plant fermentation broth particle structure 40, the protective outer layer 401 wherein the algae is selected from the group consisting of Penicillium sp., Auricularia auricula, kelp and mixtures thereof.

The novel microstructure 40 of the Rutaceae plant fermentation broth, the protective outer layer 401 structure can retain the fat-reducing, weight-loss, anti-aging and anti-wrinkle effect substances of the Rutaceae plant fermentation broth, and supplement the glycosides lost during the fermentation process. Improve the utilization rate of the human body and effectively improve the efficiency. Even if the output has not been improved, the function of the active ingredients can also achieve the same effect, even better than the efficiency of the natural structure.

The novel rutaceae plant fermentation broth particle structure 40 can be manufactured in the form of powder packs, drinks, capsules or lozenges for users to choose according to their needs.

Example 3: Experimental grouping of Rutaceae composite crystal 40 (ie, microstructure 40 of Rutaceae plant fermentation broth)

The following experiments are divided into 3-4 groups:

(1) Rutaceae water extract-Rutaceae (orange, lemon, lime, kumquat) was subjected to hot water extraction for 60 minutes.

(2) Rutaceae fermentation broth-Naturally fermented for 60 days by adding lactic acid bacteria Lactococcus acidophillus to rutaceae (orange, lemon, lime, kumquat).

(3) Rutaceae composite crystal 40 (that is, the microstructure 40 of Rutaceae plant fermentation broth)-with Rutaceae plant 10 (orange, lemon, lime, kumquat) added to lactic acid bacteria Lactococcus acidophillus for 30 days, and then yeast Saccharomycopsisfibufigera is fermented for 30 days, and finally mixed and fermented with mushroom 20 or algae 20 (shiitake, black fungus, filamentous algae, sea fungus) for 30 days.

(4) Necessary standard or control group

Composition analysis (flavonoids and amino acids) are shown in Table 1 below

After the fermentation process, the content of flavonoids in rutinaceae plants' active compounds did not change much, and the amino acids tended to increase. There was no significant difference in the content of ingredients in the process, mainly due to structural changes.

Example 4: UCP-1 brown adipose tissue protein test of Rutaceae composite crystal 40 (that is, microstructure 40 of Rutaceae plant fermentation broth)

Divide the following experiments into 6 groups of samples:

(1) 0.25% Rutaceae water extract

(2) 0.125% Rutaceae water extract

(3) 0.25% Rutaceae fermentation broth

(4) 0.125% Rutaceae fermentation broth

(5) 0.25% rutaceae composite crystal

(6) 0.125% Rutaceae composite crystal

Using 3T3-L1 preadipocytes as a model, after co-cultivation with 6 groups of samples, the content of UCP-1, a specific protein in brown adipose tissue, was tested. Conversion into brown adipocytes can increase the fat burning effect and increase the basal metabolic rate.

3T3-L1 preadipocytes were co-cultured with 6 groups of samples and transformed into brown adipocytes (the UCP-1 content increased) can increase the fat burning effect: the results (as shown in Figure 3) showed that Rutaceae composite crystal 40 showed good performance Transformation effect, with rut extract, rut fermentation liquid In comparison, the conversion rate of 0.25% rutaceae composite crystal 40 can reach 3 times in 8 hours. Moreover, the Rutinaceae composite crystals can be continuously transformed for up to 8 hours, and the Rutaceae extract and Rutin fermentation broth can only last 6 hours. Therefore, after the fermentation process, the rutaceae efficacy can be improved and the fat cell browning ability can be improved.

Example 5: Rutaceae composite crystal 40 (that is, microstructure 40 of Rutaceae plant fermentation broth) inhibits the ability of preadipocytes

Divide the following experiments into 13 groups of samples:

(1) Control group

(2) 0.125% Rutaceae water extract

(3) 0.25% Rutaceae water extract

(4) 0.5% Rutaceae water extract

(5) 1% Rutaceae water extract

(6) 0.125% Rutaceae fermentation broth

(7) 0.25% Rutaceae fermentation broth

(8) 0.5% Rutaceae fermentation broth

(9) 1% Rutaceae fermentation broth

(10) 0.125% Rutaceae composite crystal 40

(11) 0.25% Rutaceae composite crystal 40

(12) 0.5% Rutaceae composite crystal 40 (that is, the microstructure 40 of Rutaceae plant fermentation broth)

(13) 1% rutaceae composite crystal 40 (that is, microstructure 40 of rutaceae plant fermentation broth)

NIH-3T3 fibroblasts were used as the toxicity test cell model to confirm that the samples were not toxic to the cells, and then 3T3-L1 preadipocytes were used as the model, co-cultured with the samples for 24 or 48 hours, and the cell survival rate was tested by MTT assays. Inhibit the differentiation of preadipocytes and reduce the generation of adipocytes.

Toxicity test cell results: The results (shown in Figures 4 and 5) show that after 24 hours and 48 hours of culture, the Rutaceae water extract, Rutaceae fermentation broth, and Rutaceae composite crystal 40 (that is, the microstructure of the Rutaceae plant fermentation broth) 40) There is no poisonous killing. The survival rate of fibroblasts exceeds 85%, with no adverse effects.

The result of inhibiting the ability of preadipocytes: The results (shown in Figures 6 and 7) show that after 24 hours and 48 hours of culture, the inhibitory effect of preadipocytes is: Rutaceae composite crystal 40 (that is, microstructure 40 of Rutaceae plant fermentation broth) >Rutaceae water extract>Rutaceae fermentation broth. It is known from Figure 6 that the general fermentation method cannot improve the functional ingredients in citrus. The Rutaceae composite crystal 40 (that is, the microstructure 40 of Rutaceae plant fermentation broth) exhibited significant ability to inhibit preadipocytes. The 24-hour and 48-hour survival rates were 65% and 50%, effectively reducing the formation of adipocytes.

Example 6: Antioxidant ability test of Rutaceae composite crystal 40 (that is, microstructure 40 of Rutaceae plant fermentation broth)

Divide the following experiments into 6 groups of samples:

(1) 0.25% Rutaceae water extract

(2) 0.125% Rutaceae water extract

(3) 0.25% Rutaceae fermentation broth

(4) 0.125% Rutaceae fermentation broth

(5) 0.25% Rutaceae composite crystal 40

(6) 0.125% Rutaceae composite crystal 40

Using common free radicals: DPPH, ABTS+ and NBT as indicators, in-vitro tests were conducted, and 6 groups of samples with different concentrations were added to test whether the samples have the ability to scavenge free radicals.

Antioxidant ability results: The results (shown in Figure 8) show the ability to scavenge DPPH free radicals: Rutaceae composite crystal 40> Rutaceae water extract> Rutaceae fermentation broth, where the Rutaceae composite crystal achieves 63% scavenging effect; results (As shown in Figure 9) Ability to remove ABTS+ free radicals: Rutaceae composite crystal 40> Rutaceae water extract> Rutaceae fermentation broth, in which the Rutaceae composite crystal achieves 57% scavenging effect; results (shown in Figure 10) Ability to scavenge NBT free radicals: Rutaceae composite crystal 40> Rutaceae water extract> Rutaceae fermentation broth, wherein the Rutaceae composite crystal 40 achieves 90% scavenging effect.

Example 7: Inhibition of tyrosinase activity of Rutaceae composite crystal 40 (ie, microstructure 40 of Rutaceae plant fermentation broth)

Divide the following experiments into 8 groups of samples:

(1) 0.125% Kojic acid

(2) 0.25% Kojic acid

(3) 0.125% Rutaceae water extract

(4) 0.25% Rutaceae water extract

(5) 0.125% Rutaceae fermentation broth

(6) 0.25% Rutaceae fermentation broth

(7) 0.125% Rutaceae composite crystal

(8) 0.25% Rutaceae composite crystal

Tyrosine is catalyzed by tyrosinase to produce dopa, which is a melanin precursor. Test whether the 8 groups of samples can inhibit the activity of tyrosinase and reduce the production of melanin precursors.

Inhibition of tyrosinase activity results: the results (as shown in Figure 11) show that each sample is concentration-dependent; inhibition of tyrosinase activity test: kojic acid> rutaceae composite crystal 40> rutaceae water extract> rut Branch fermentation broth; wherein the Rutaceae composite crystal 40 reaches 76% inhibition rate, reducing the production of melanin precursors.

Example 8: MMP-2 protein expression test of Rutaceae composite crystal 40 (that is, microstructure 40 of Rutaceae plant fermentation broth)

Divide the following experiments into 9 groups of samples:

(1) Control group

(2) 0.125% Vitamin (Vitamin)

(3) 0.25% Vitamin (Vitamin)

(4) 0.125% Rutaceae water extract

(5) 0.25% Rutaceae water extract

(6) 0.125% Rutaceae fermentation broth

(7) 0.25% Rutaceae fermentation broth

(8) 0.125% Rutaceae composite crystal 40

(9) 0.25% Rutaceae composite crystal 40

MMPs are metalloproteinases, which are present in the dermal cell matrix and mainly act to microlyse collagen. Pro MMPs are in the unactivated state of MMPs. If the concentration of Pro MMPs increases, it means that the activation of MMPs is reduced and collagen maintenance is achieved.

MMP-2 protein performance results: the results (as shown in Figure 12) show that each sample is concentration-dependent; Pro MMP-2 protein concentration: Rutaceae composite crystal 40> Vitamin C> Rutaceae water extract> Rutaceae Fermentation broth, in which the Rutaceae composite crystal 40 increases the concentration of Pro MMP-1, to avoid collagen cleavage caused by the activation of MMP-2.

Example 9: Obese mouse experiment of Rutaceae composite crystal 40 (that is, microstructure of fermented broth of Rutaceae plant)

Divide the following experiment into 2 groups of samples:

(1) Control group (without giving Rutaceae composite crystals)

(2) Rutaceae composite crystal 40

The mice were fed with high fat feed to induce obesity, and the rutinaceous compound crystal was also given to monitor the weight change rate of the mice.

Obese mice test results: The results (shown in Figure 13) show The weight growth rate of mice fed with feed increased with time, and the weight gain rate reached 39% on the 19th day; mice with rutinaceous compound crystals can reduce the rate of weight change and reduce the weight gain rate to 8.

40‧‧‧Particulate structure of Rutaceae plant fermentation broth

401‧‧‧Protect the outer layer

402‧‧‧Inner layer

Claims (7)

A microstructure of fermented broth of Rutaceae plant includes, in order from the center to the outside: an inner layer; and a protective outer layer, wherein the protective outer layer covers the inner layer; wherein the inner layer contains a fermented broth of Rutaceae plant; The protective outer layer contains a small molecule polysaccharide molecule contained in a mushroom and an algae after special fermentation. The microstructure of the fermentation broth of Rutaceae as described in item 1 of the patent application, wherein the diameter of the inner layer is between 10 ^-5 ~10 ^-4 mm. The microstructure of the fermentation broth of Rutaceae plant as described in item 1 of the patent application scope, wherein the particle size of the protective outer layer is between 10 ^-4 ~10 ^-3 mm. The microstructure of the Rutaceae plant fermentation broth as described in item 1 of the patent application scope, wherein the Rutaceae plant contained in the Rutaceae plant fermentation broth is selected from the group consisting of orange, lemon, lime, kumquat and mixtures thereof Group. The microstructure of the fermentation broth of Rutaceae plant as described in item 1 of the patent application scope, wherein the mushrooms in the protective outer layer are selected from the group consisting of mushrooms, black fungus, white fungus and mixtures thereof. The microstructure of the fermentation broth of Rutaceae plant as described in item 1 of the scope of patent application, wherein the algae in the protective outer layer is selected from the group consisting of Penicillium sp., Auricularia auricula, kelp and mixtures thereof. The microstructure of the fermentation broth of Rutaceae plant as described in item 1 of the patent scope, wherein the rut The microbial structure of the branch plant fermentation broth contains powder packs, drinks, capsules or lozenges for users to choose according to their needs.

Images