WO2007030975A1

WO2007030975A1 - New usage of tremella fuciformis heteropolysaccharide or extract thereof

Info

Publication number: WO2007030975A1
Application number: PCT/CN2005/001574
Authority: WO
Inventors: Guiyun Liu; Feng Luo; Bin Luo
Original assignee: Shanghai Wenda Biotech Inc.
Priority date: 2005-09-16
Filing date: 2005-09-26
Publication date: 2007-03-22
Also published as: JP5498698B2; JP2009507863A; CN1931879A; CN100484964C

Abstract

The usage of Tremella fuciformis heteropolysaccharide or T. fuciformis heteropolysaccharide extract in preparation of medicaments, food or functional food for promoting the growth and/or propagation of beneficial bacterium, in preparation of medicaments, food or functional food for ameliorating constipation, in preparation of medicaments, food or functional food of cosmetologic activity, and in preparation of accelerants for promoting the growth and/or propagation of beneficial bacterium in vitro.

Description

New use of Tremella fuciformis or its extracts

The present invention relates to the use of a heteropolysaccharide; in particular, to the use of a Tremella heteropolysaccharide or an extract thereof for promoting the growth and/or reproduction of probiotics, and the use thereof for improving constipation function and detoxifying beauty function. technical background

Bifidobacteria are physiological bacteria in the human body. They are Gram-positive, non-spore-free, non-exercise, and strictly anaerobic. They are dominant in the human intestine. The ultimate survival lies in the human body. It is closely related to human health. It is a kind of human body. A beneficial bacteria with many important physiological functions. With the rapid development of microbiology, the clinical health effects of bifidobacteria have gradually been recognized and accepted. Human physiology, immunity, nutrition, digestion, anti-tumor, anti-aging, drug efficacy, etc. are all inseparable from the microbial population carried by the human body, and the bifidobacteria in the human intestinal tract has disease treatment, disease-free anti-aging and health care. The role. For example, bifidobacteria can form a physiological barrier, prevent the colonization and invasion of pathogenic bacteria, protect the host from pathogenic bacteria, improve protein metabolism, provide a variety of B vitamins, promote calcium and iron absorption, improve immunity, and enhance Anti-tumor ability and other effects. If the deficiency of bifidobacteria in the human body means that the human body has a health crisis, it needs to be replenished in time.

Lactobacillus is also an important physiological bacterium in the human body. It is Gram-positive, non-spore-free, non-exercise, and strictly anaerobic. It has various physiological functions. Lactobacillus maintains the balance of the intestinal and genital flora and plays a key role in removing certain carcinogens or preventing carcinogenesis. The reduction of lactobacilli may lead to dysregulation of the intestinal flora, and the pathogens multiply in large numbers, and some bacterial enzymes in the metabolites can catalyze the conversion of carcinogenic precursor substances to carcinogens. Lactobacillus can restore the balance of intestinal flora, resist the growth of pathogenic bacteria in the intestinal tract, inhibit the production of bacterial enzymes and reduce its activity. It can also be combined with carcinogens to excrete with feces, reducing its chances of formation, activation and retention.

Prebiotics are a class of non-digestible substances, but can be used as a substrate by the normal intestinal flora, which can selectively stimulate the growth and reproduction of one or several beneficial bacteria in the intestine, inhibit the growth of harmful bacteria, and host Health has beneficial effects, such as stachymile, oligofructose (including isomalto-oligosaccharides, galactose oligosaccharides, fructooligosaccharides, etc.. Because these substances can promote the growth and / or reproduction of bifidobacteria, it is also called As a "bifidus factor", oligofructose is an internationally recognized bifidus factor, but they are all oligosaccharides with a relatively small molecular weight. Promote body by taking prebiotics. The growth and/or reproduction of probiotics can further improve the function of the human digestion and absorption system, help to improve constipation and promote human metabolism, and the toxins in the body are discharged in time, and the effect of beauty and beauty can be achieved.

Tremella faciformis Berk, also known as white fungus, mulberry goose, and Qing dynasty. It belongs to Tremellales, Tremellacease, Tremella. It has the effects of nourishing yin and moistening the lungs, benefiting Qi and blood. Tremella fuciformis is an acidic heteropolysaccharide extracted from the fungus fruit body (or its microbial fermentation broth). Since ancient times, China has called white fungus high-grade supplements and medicines. The best for medicine and food is the court's finest tonic food.

With the continuous development of science and the continuous improvement of technical means, people have re-recognition of Tremella. At home and abroad, a lot of researches have been carried out on the main component of Tremella fuciformis, Tremella fuciformis, which indicates that Tremella fuciformis has improved immunity and resistance. Tumor, anti-aging, blood sugar lowering, blood lipids, protein nucleic acid synthesis, antithrombotic and other pharmacological effects. Japan has applied it extensively to health products, cosmetics, and pharmaceuticals. The development and application of Tremella has aroused great interest in the scientific community, and has received extensive attention in the international pharmaceutical and cosmetic fields, and has become a hot topic in current research and development.

However, the composition and structure of the Tremella polysaccharides and the corresponding functions have rarely been reported so far. The applicant uses a special separation and purification process to refine the active polysaccharide in Tremella fuciformis, which is identified by modern analytical techniques. The structure of the Tremella heteropolysaccharide is α - ( 1-3 ) mannose as the main chain, mainly by weight percentage 70- 93% neutral total sugar, 6-28% glucuronic acid, 0.1-3% binding protein; the average molecular weight of the Tremella heteropolysaccharide is 85-1.6 million Daltons, of which the molecular weight is greater than 6000 Daltons. 90% (by weight) or more, as described in the patent application No. 200510023427.4 filed on Jan. 18, 2005, and the name of the product is WSK; the patent application also discloses the white fungus heteropolysaccharide and The extract has the functions of moisturizing and smoothing the skin and mucous membrane, enhancing the sense of lubrication, anti-oxidation aging, anti-allergy, thickening and stability. However, to date, there has been no report on the function and use of Tremella heteropolysaccharide to promote the proliferation and improvement of constipation in the intestinal and digestive systems, as well as the function and use of detoxification. Summary of the invention

The object of the present invention is to provide a silver fungus heteropolysaccharide or a tremella polysaccharide extract of the trade name WSK in the patent application No. 200510023427.4 for the preparation of a pharmaceutical, food or drink or health supplement for promoting the growth and/or reproduction of probiotics. Use in.

Another object of the present invention is to provide a Tremella heteropolysaccharide or a Tremella polysaccharide extract of the trade name WSK in the patent application No. 200510023427.4 for the preparation of a medicine, a food or drink or a health care product having an improved constipation function. Use in the product.

A further object of the present invention is to provide a silver fungus heteropolysaccharide or a silver fungus heteropolysaccharide extract having the trade name WSK in the application No. 200510023427.4, in the preparation of a medicine, a food or drink or a health care product having a detoxifying beauty function. use.

The structure of the Tremella heteropolysaccharide according to the present invention is mainly composed of α-(1-3) mannose, mainly composed of 70-93% by weight of neutral total sugar, 6-28% of glucuronic acid, 0.1- 3% of the binding protein composition; the average molecular weight of the Tremella heteropolysaccharide is 85-1.6 million Daltons, wherein more than 90% by weight of the molecular weight is greater than 6000 Daltons. Wherein, the neutral total sugar mainly comprises the following content of glycosyl: 35-60% mannose, 10-25% fucose, 8-18% xylose and 5-15% glucose, the above content is white fungus Heteropolysaccharide weight meter.

The side chain of the tremella heteropolysaccharide has a plurality of different combinations of changes, one of which has been determined by X-ray diffraction as a structural unit represented by the following structural formula I -

→3) - a -Manp- (1 -3) - α - Manp- (1→3) -α-Manp- (1→3) -a - Manp- (1—3) - (continued down)

2 2 2

t † t

1 1 1

β -Xylp β -GlcAp β -GlcAp

(up to the top)

α - (1→3)

(I)

The structural unit has a molecular weight of 2578.5 or 2724.7. The sugar structure is composed of α-(1-3) mannose (Manp) as the main chain skeleton, and two β-(1-2) xylose (Xylp) and three β-glucose are attached to the two carbon atoms. Aldehydic acid (GlcAp), 1 or 2 fucose (Fuc), 1 mannose is a side chain.

The Tremella heteropolysaccharide extract of the present invention mainly comprises 80-98% by weight of the above-mentioned Tremella heteropolysaccharide, 0.1-3.3% of free protein and 0.5-8% of ash; the average molecular weight of the Tremella heteropolysaccharide extract is 75-1.4 million Daltons, of which 75-98% by weight of molecular weight greater than 6000 Daltons.

The Tremella heteropolysaccharide extract of the present invention can be obtained by the following preparation method - 1) The raw material of the tremella fruit body is stirred with water at 80-100 ° C to obtain a crude extract ^,

2) The crude extract obtained in the step 1) is filtered, and the filtrate finally passes through a filter having a pore diameter of 0.1-1 μm;

3) adding ethanol to the filtrate obtained in the step 2) to make the final volume concentration of the ethanol 50-85%, and collecting the precipitate;

4) The precipitate obtained in the step 3) is dried.

Wherein, the raw material of the white fungus fruit body is a dry raw material of the tremella fruit body, and the dried tremella fruit material is soaked, expanded and cleaned with water (including a yellow-brown base), 20-100 times each time, and multiple times of extraction. Not more than 200 times by weight of the dried fermented fruit body material amount of water is stirred and extracted at 80-100 ° C to obtain a crude extract.

The raw material of the white fungus fruit body may also be a raw material of fresh white fungus fruit body, fresh frozen fungus fruit raw material or frozen fresh white fungus fruit material, and the raw material of the white fungus fruit body is removed to a yellow-brown base and cleaned, and each time 2- 15 times, multiple extractions, no more than 30 times (by weight) of the amount of water of the raw material of the fungus fruit body is stirred and extracted at 80-100 ° C to obtain a crude extract.

Another preparation method of the Tremella heteropolysaccharide extract of the present invention comprises the following steps -

1) According to the conventional microbial fermentation method (Reference 1 research on the deep-fermentation conditions of Tremella fuciformis, Wu Dakang et al; Food Science; 2002, Vol.23, No. 64; 2 Study on the separation method of Tremella fuciformis, Zhang Bingchi; Chinese edible fungus; Vol .13, No.3; 22) The obtained white fungus fermentation broth is heated to 80-100 ° C, incubated for 0.5-2 hours, filtered, and the supernatant is finally passed through a filter having a pore diameter of 0.1-1 μ m to obtain a filtrate;

2) adding ethanol to the filtrate obtained in the step 1) to make the final volume concentration of the ethanol 50-85%, collecting the precipitate;

3) The precipitate obtained in the step 2) is dried.

The method for preparing the Tremella heteropolysaccharide of the present invention comprises the step of further purifying the extract obtained by the above preparation method.

Among them, the purification was carried out by the Sevag method and the dialysis method. Of course, as is known to those skilled in the art, other polysaccharide purification methods, such as chromatographic separation methods, can also produce the Tremella heteropolysaccharide of the present invention.

The probiotic may be a variety of probiotics, mainly bifidobacteria and / or lactobacilli; preferably bifidobacteria and / or lactobacilli in the digestive system (such as the intestine), or the reproductive system (such as Lactobacillus in the reproductive tract).

The Bifidobaterinm is an obligate anaerobes of irregular shape, and there are 32 species, preferably Bifidobacterium bifidum, Bifidobacterium adolescentis or Bifidobacterium infantis, more preferably Bifidobacterium bifidum; Bifidobacterium bifidum The best is Bifidobacterium ATCC 2952. The Lactobacillus is a Gram-positive Bacillus-free bacterium, and there are 56 species, preferably Lactobacillus delbrueckii, Lactobacillus casei, and Lactobacillus brevis, more preferably Lactobacillus delbrueckii; Preferably, it is a Lactobacillus delbrueckii subsp. bulgaricus, more preferably Lactobacillus bulgaricus ATCC 11480.

The medicine may be various dosage forms in the prior art, such as powders, tablets, capsules (including soft and hard capsules), liquids, suspensions, emulsions, oral liquids, jelly or granules; The food and drink includes drinks and foods, and is a drink or food containing a concentrate and/or a dried product of Tremella heteropolysaccharide or Tremella heteropolysaccharide extract, and the application of the food or drink is not limited, and may be various dosage forms in the prior art. If it can be used in powders, tablets, capsules (including soft and hard capsules), granules, granules, jelly, beverages, dairy products, yoghurt or snacks, etc.; the health care products can be prior art Various dosage forms, such as powders, tablets, capsules (including soft and hard capsules), liquids, suspensions, emulsions, oral liquids, jelly or granules, and the like.

According to the present invention, the pharmaceutical, food or health supplement may further contain other prebiotics which promote the growth and/or reproduction of the probiotic.

The food or drink or health supplement may also contain various other edible nutrients for better nutrition and health care.

The Tremella heteropolysaccharide according to the present invention may be crude crude Tremella heteropolysaccharide containing impurities or pure Tremella polysaccharide.

A further object of the present invention is to provide a use of the Tremella heteropolysaccharide or Tremella polysaccharide extract of the trade name WSK in the patent application No. 200510023427.4 for promoting the growth and/or propagation of probiotics in vitro.

In the present invention, the action or function of the promoter is similar to the bifidus factor, and the difference is that: the bifidus factor is a substance that promotes the growth and/or reproduction of bifidobacteria, and the promoter promotes probiotics. (including bifidobacteria) substances that grow and/or reproduce. The accelerator according to the present invention is added to medicines, foods, and health care products, and can promote the growth and reproduction of probiotics in the body after administration; and the promotion of the present invention is added to medicines, foods, and health care products containing probiotics. Agents can enhance the efficacy of such products. The accelerator of the present invention can be used, for example, in a culture medium to culture probiotics in vitro, so that more beneficial bacteria can be cultured in a faster manner, thereby reducing costs.

The probiotics may be various probiotics, mainly bifidobacteria and/or lactobacilli.

The Bifidobacterium is preferably Bifidobacterium bifidum B. bifidum, more preferably Bifidobacterium ATCC 2952; The Lactobacillus is preferably Lactobacillus delbrueckii, more preferably the Lactobacillus delbrueckii subsp. bulgaricus, most preferably Lactobacillus bulgaricus ATCC 11480.

The positive progress of the present invention is that: Tremella fuciformis extract and Tremella fuciformis (WSK) promote the growth and/or reproduction of probiotics, such as direct regulation of human intestinal bacteria, proliferation of bifidobacteria and/or lactic acid in vivo. Bacillus, will have a broader development prospects. Moreover, the proliferative effect of Tremella polysaccharides on probiotics such as bifidobacteria and lactobacilli will be further developed as a promoter (similar to the function or action of bifidus factors) and promote the development of microecology. The medicine, food or health care product containing the extract of Tremella heteropolysaccharide or Tremella heteropolysaccharide of the present invention has the function of improving constipation and the function of detoxifying beauty and beauty after oral administration, which excretes toxins in the body and absorbs nutrients in the silver fungus heteropolysaccharide Can play a role in beauty and beauty. DRAWINGS

Figure 1 is a comparison of the effects of sugar-free, oligosaccharide and tremella heteropolysaccharide on the growth and reproduction of Bifidobacterium ATCC 2952; Figure 2A is a photograph of the growth and reproduction of Bifidobacterium ATCC 2952 in control group 1;

2B is a photograph showing the growth and reproduction of Bifidobacterium ATCC 2952 in the experimental group (Tremella heteropolysaccharide group); FIG. 3 is a comparison chart showing the effects of sugar-free, oligosaccharide and Tremella heteropolysaccharide on the growth and reproduction of Lactobacillus bulgaricus ATCC 11480;

Figure 4A is a photograph showing the growth and reproduction of Lactobacillus bulgaricus ATCC 11480 of Control Group 1;

Fig. 4B is a photograph showing the growth and reproduction of Lactobacillus bulgaricus ATCC 11480 in the experimental group (the Tremella heteropolysaccharide group); Fig. 5 is a schematic view showing the improvement effect of the different concentrations of Tremella fuciformis on constipation;

Figure 6 is a schematic diagram of the lubricating effect of different concentrations of Tremella heteropolysaccharide solution on constipation patients and normal human stools. Figure 7 is a schematic diagram showing the effect of different concentrations of Tremella fuciformis extract (WSK-1) on constipation; Figure 8 is the lubrication effect of different concentrations of Tremella fuciformis extract (WSK-1) solution on constipation patients and normal stools Schematic diagram. detailed description

The desired Tremella heteropolysaccharide extract and Tremella heteropolysaccharide (WSK) are prepared according to the preparation method in the applicant's patent application No. 200510023427.4, which is described in detail as follows: Example 1

Soak the dried white fungus fruit body with water, swell it, and remove it to the yellow-brown base and clean it; put the washed raw material into the extraction tank, add 100 times the dry water, stir and extract for 4 hours at 100 ° water temperature, then Filtration, the supernatant finally passed through a filter with a pore size of Ol-ΐμιη; under uniform agitation, the filtrate and ethanol were thoroughly mixed, and the final concentration of ethanol in the mixed liquid was 85%. The precipitate was collected, dehydrated with pure ethanol, and the precipitate was collected. ; -40 ° C freeze-dried, pulverized Tremella heteropolysaccharide extract (WSK-1). The product yield is 20% (based on the weight of dry white fungus), and the purity of the Tremella polysaccharide is 80%.

Example 2

Store the fresh-keeping frozen white fungus on the yellow-brown base and clean it; put the washed raw material into the extraction tank, add 15 times of fresh water, stir and extract at 10CTC water temperature for 4h, then filter, the supernatant finally passes a filter with a pore diameter of 0.1-1 μm; the filtrate is heated and concentrated to obtain a concentrated solution; under uniform stirring, the filtrate and ethanol are thoroughly mixed, and the final concentration of ethanol in the mixed liquid is 85%, and the precipitate is collected, using pure ethanol. Dehydration and decolorization, collection of precipitate; 80 ° C hot air drying, pulverization of Tremella heteropolysaccharide extract. Product yield 2.1% (based on the weight of fresh white fungus).

Example 3

The Tremella fermentation broth obtained by microbial fermentation is heated at 100 ° C for 0.5 h, filtered conventionally, and the filter residue is discarded. The supernatant is finally passed through a filter having a pore diameter of 0.1-1 μm; the filtrate and the ethanol are thoroughly mixed under uniform stirring. The final concentration of ethanol in the mixed liquid was 85%, the precipitate was collected, dehydrated by dehydration with 100% ethanol, and the precipitate was collected; -40 Ό freeze-dried, and pulverized to obtain a white fungus heteropolysaccharide extract. Product yield 0.55% (based on the weight of the fermentation broth).

Example 4

The trehalose polysaccharide extracts of Examples 1 to 3 were dissolved in 600 times by weight of deionized water, and after dissolution, 200 mL of a mixed solution of chloroform: n-butanol = 4:1 (volume ratio) was added, and after thorough mixing, The centrifuge was centrifuged (4000 r/min, 30 minutes) and the supernatant was collected. The supernatant after centrifugation was mixed with the above-mentioned chloroform-n-butanol mixed solution at a ratio of 3:1 (volume ratio), and centrifuged (4000 r/min, 30 minutes) with a centrifuge, and the supernatant was collected. Repeat the above operation and repeat the treatment until the OD value of the aqueous phase at 280 nm (using the 752 UV grating spectrophotometer, Shanghai Precision Scientific Instrument Factory) is no longer falling. The aqueous phase was concentrated under reduced pressure to one-third, and the concentrate was thoroughly mixed with 3 times (volume ratio) of 95% ethanol in the aqueous phase, and centrifuged at 4000 r/min for 30 minutes to obtain a precipitate. The precipitate was dissolved in 100 times by weight of deionized water, centrifuged, and insoluble, and the supernatant was placed in a 30 cm long dialysis bag (dialysis bag can pass 5000 molecular weight or less) Qualitatively perform dialysis (both ends are placed in a bottle filled with deionized water and rinsed with deionized water for 12 hours) to collect 5000 molecular weight or higher substances. The dialyzed supernatant was thoroughly mixed with 3 times (volume) pure B, and centrifuged again at 4000 r/min for 30 minutes to obtain a precipitate. The precipitate was dehydrated with absolute ethanol and dried under vacuum at 60 ° C to obtain pure silver ear polysaccharide. (WSK).

Effect Example 1 Tremella heteropolysaccharide (WSK) promotes the growth and/or reproduction of bifidobacteria

Sample Name: Tremella heteropolysaccharide (WSK) obtained in Example 4, oligofructose (standard product provided by Wuxi University of Light Industry Food Institute)

Test strain: Bifidobacteria ATCC 2952, purchased from the American Type Culture Collection

Medium: MRS Broth Medium

Sterilization method: Sterilize at 90 ° C for 30 minutes

The experimental treatment was divided into the following three items - control group 1: medium 6.3 mL + bacterial solution 0.3 mL

Control group 2: medium 6.0 mL + oligofructose 0.3 mL + bacterial solution 0.3 mL (the mass ratio of oligofructose: cultivating medium in the medium is 1: 80)

Experimental group: Medium 6.0 mL + Tremella heteropolysaccharide sample solution 0.3 mL + bacterial solution 0.3 mL (The white fungus heterozygous in the medium: sugar: medium mass ratio is 1: 80)

Experimental method: liquid culture method

Please give the procedure for cultivating bifidobacteria: Add vitamin 2.0 to 0.1 wt% with anaphylatobacterium basal medium (Shanghai Institute of Biological Products), prepare a liquid medium, and culture in a conventional incubator at 37 °C for 48 hours.

Detection method: Liquid quantitative limiting dilution method.

Note: Quantitative counting of bacteria in liquid culture method 560nm OD value: 1.0 OD value = 6X 108 bacteria / mL, 2.0 OD value = 18 X 108 bacteria / mL, 3.0 OD value = 24X 108 bacteria / mL.

The experimental results show that the addition of Tremella heteropolysaccharide (1:80 dilution) to the liquid medium can promote the growth and reproduction of Bifidobacterium, and the effect is remarkable. As can be seen from Fig. 1, compared with the sugar-free medium in the control group 1, the Tremella fuciformis polysaccharide can significantly increase the amount of Bifidobacterium in the fermentation broth, and the promotion of the growth and reproduction of Bifidobacterium by the Tremella polysaccharides Even more than the oligofructose currently used as a prebiotic in the market. Moreover, Bifidobacterium ATCC 2952 The culture time in the MRS broth medium was advanced from 48 hours to 24 hours; and by smear staining, compared with the control group 1, the bacteria in the experimental group were enlarged and thick, and the density of the bacteria in the same field of view was markedly Increase 3⁄4, see Figure 2A and Figure

2Bo ”

Effect Example 2 Tremella heteropolysaccharide (WSK) promotes the growth and/or reproduction of lactobacilli

Test strain: Lactobacillus ATCC 11480, purchased from the Institute of Microbiology, Chinese Academy of Sciences. Medium: MRS Broth Medium

Medium sterilization method: Sterilize at 9CTC for 30 minutes

The experimental processing is divided into the following three items:

Control group 1 : medium 6.3 mL + bacterial solution 0.3 mL

Control group 2: Medium 6.0 mL + oligofructose 0.3 mL + bacterial solution 0.3 mL (The ratio of oligofructose: medium in the medium was 1: 80)

Experimental group: medium 6.0 mL + WSK sample solution 0.3 mL + bacterial solution 0.3 mL (the mass ratio of the silver fungus heteropolysaccharide: Pei 'nutrients in the medium is 1: 80)

Experimental method: liquid culture method

Cultivation of Lactobacillus: ₀₂ added with anaerobic bacteria basal medium (Shanghai Institute of Biological Products) vitamin K to a concentration of 0.1wt%, is made of the liquid medium, in a conventional culture incubator 37 ° C, stationary for 48 hours.

Detection method: Liquid quantitative limiting dilution method.

Note: When counting the 560nm OD value of the bacteria in the liquid culture method: 1.0 OD value = 6X 10 ⁸ bacteria / mL, 2.0 OD value = 18 X 10 ⁸ bacteria / mL, 3.0 OD value = 24 X 10 ⁸ bacteria / mL .

The experimental results show that the addition of Tremella heteropolysaccharide (1:80 dilution) to the liquid medium can promote the growth and reproduction of Lactobacillus, and the effect is remarkable. As can be seen from Fig. 3, compared with the sugar-free medium in the control group 1, the Tremella polysaccharide can significantly increase the amount of Lactobacillus in the fermentation broth, and the promotion effect of the Tremella polysaccharide on the growth and reproduction of Lactobacillus even More than the oligofructose currently used as a prebiotic in the market. Moreover, the culture time of Lactobacillus bulgaricus ATCC 11480 in MRS broth medium was advanced from 48 hours to 24 hours; and by smear staining, compared with the control group 1, the cells of the experimental group were enlarged and thick, in the same field of view. The density of the lower bacteria is significantly increased, See Figures 4A and 4B.

Effect Example 3 The effect of Tremella fuciformis (WSK) on constipation improvement in constipation patients (whether it is improved) and the oral effects on normal people and the change in skin gloss on the face after taking (testing whether it has the function of beauty and beauty) was tested.

Subjects: 16 patients with constipation, the symptoms were 3 times/week of bowel movements or dry stools, gender age is not limited, divided into 3 groups, Al, A2, A3 group, taking 0.1wt%, 0.2wt%, 0.5wt% respectively The Tremella heteropolysaccharide solution was used to observe changes in stool condition and facial skin luster. Sixteen patients with normal bowel movements were divided into three groups, Bl, B2, and B3, taking 0.1 wt%, 0.2 wt%, and 0.5 wt ^Q / respectively. The Tremella heteropolysaccharide solution was used to observe changes in stool condition and facial skin gloss.

Preparation of Tremella Heteropolysaccharide Solution: Sterile treatment to ensure the health and safety of the drinker, and to eliminate the influence of bacteria on the test results;

1) preparing a sufficient amount of 0.1 wt%, 0.2 wt%, 0.5 wt% Tremella heteropolysaccharide solution for testing, the Tremella heteropolysaccharide. The Tremella heteropolysaccharide (WSK) obtained in Example 4;

2) Preparation of bacterial test results <20cfu / g sterile Tremella heteropolysaccharide powder, preservatives selected potassium sorbate. (Additional amount 0.1~0.2wt%, refer to the Chinese food additive manual, potassium sorbate instructions and microbiological test results );

3) Preparation method: Take a small amount of distilled water and heat to boil for 10 minutes. After cooling to 70~80 °C, add potassium sorbate, dissolve and reserve. The remaining distilled water is added to the tremella heteropolysaccharide powder homogenizer and stirred to dissolve fully. Later, potassium sorbate aqueous solution is added. Adding, mixing and hooking, the whole operation process is strictly sterile;

4) Bottle (PE bottle) cleaning method: 75% alcohol soaking, clean room 24 hours to dry;

5) Requirements: Sterile, dry, sealed, used in tank assembly of the Tremella heteropolysaccharide solution.

Experimental methods: Patients with constipation and normal bowel movements were treated with Tremella polysaccharide solution twice a day for 2 hours after dinner and 2 days after dinner;

Groups A1 and B1: Each dose is 100mL, 2 times a day;

Group A2 and B2: Each dose is lOOmL, 2 times a day;

Groups A3 and B3: Each dose is 100mL, 2 times a day;

After 7 days of continuous administration, the number of bowel movements was counted by the user, and the presence or absence of a body abnormality and the change in the skin gloss of the face observed 7 days ago were recorded. The effect of Tremella fuciformis on improving constipation and the improvement of skin gloss in constipation patients are shown in Table 5. The improvement effect of different concentrations of Tremella polysaccharides on constipation is shown in Fig. 5. The effect of Tremella fuciformis on the bowel movement of normal stools and the skin of the users The improvement of gloss is shown in Table 2. The lubrication effect of different concentrations of Tremella heteropolysaccharide solution on constipation patients and normal stools is shown in Fig. 6.

Table 1 Effect of Tremella Heteropolysaccharide on Constipation

Note: Invalid: The number of stools does not change

Improvement: Increased number of bowel movements 1~2 times

Significant improvement: The number of bowel movements increased by 2 to 4 times, and the stool was obviously lubricated

Table 2 Effect of Tremella polysaccharides on laxative in normal people Group B1 B2 B3 Tremella heteropolysaccharide 0.1% 0.2% " 0.5%

Concentration

Number of people 5 people 6 people 5 people

Weekly number of bowel movements unchanged: 4 people unchanged: 5 people unchanged: 4 people

Increase: 1 person Increase: 1 person Increase: 1 person Stool Lubrication 60% 50% 100%

Cause constipation None No No

Other discomfort

Facial skin gloss No change: 2 people No change: 2 people No change: 1 person

Observed Gloss slightly increased: 3 people slightly increased gloss: 2 slightly increased gloss: 1 person significantly increased gloss: 0 people significantly increased luster: 3 people significantly increased gloss: 2

People

The experimental results show that: 1) Tremella fuciformis has the function of lubricating stool and improving constipation;

2) The improvement effect increases significantly with the increase of the concentration of the silver fungus heteropolysaccharide;

3) The effect of gender and age on the improvement effect is not obvious;

4) There is no side effect on the body of patients with constipation, and it will not cause bloating, abdominal pain and other feelings;

5) Tremella heteropolysaccharide has no side effects on normal people, no bloating, and the stool is relatively lubricated;

6) Tremella fuciformis has a beauty-enhancing effect of improving skin luster, and the improvement effect is markedly increased with the increase of the concentration of the tremella heteropolysaccharide.

Effect Example 4 The effect of the extract of Tremella fuciformis on constipation in constipation patients (whether it is improved) and the oral effect on normal people and the change in skin gloss on the face after taking (whether or not it has a beauty and beauty function) were tested.

Subjects: 18 patients with constipation, the symptoms were bowel movements 3 times / week or dry stool, gender age is not limited, divided into 3 groups, Cl, C2, C3 group, taking 0.1wt%, 0.2wt%, 0.5wt% Tremella heteropolysaccharide extract solution, observe changes in stool condition and facial skin luster; 18 normal bowel movements, divided into 3 groups, Dl, D2, D3 group, respectively, taking 0.1wt%, 0.2wt%, 0.5wt% Tremella heteropolysaccharide extract solution, observe stool status and Changes in the luster of the skin on the face.

Preparation of Tremella Heteropolysaccharide Extract Solution: Sterile treatment, ensuring the health and safety of the user, and eliminating the influence of bacteria on the test results;

1) preparing a sufficient amount of 0.1 wt%, 0.2 wt%, 0.5 wt% of the Tremella polysaccharide extract solution for the test, the Tremella heteropolysaccharide extract is the Tremella fuciformis extract (WSK-1) obtained in Example 1, The purity of Tremella fuciformis polysaccharide is 80%;

2) Preparation of bacterial test results <20db / g sterile Tremella heteropolysaccharide extract powder, preservatives selected potassium sorbate (addition 0.1 ~ 0.2wt%, refer to the Chinese food additive manual, potassium sorbate instructions and microbial test Result);

3) Preparation method: Take a small amount of distilled water and boil for 10 minutes. After cooling to 70~80 °C, add potassium sorbate, dissolve and reserve. The remaining distilled water is added to the white fungus heteropolysaccharide extract powder homogenizer and fully dissolved. The potassium sorbate aqueous solution is added, mixed evenly, and the whole operation process is strictly sterile;

5) Requirements: Sterile, dry, sealed, and used in the preparation of cans of Tremella heteropolysaccharide extract solution.

Experimental method: Patients with constipation and normal bowel movements took the silver ear polysaccharide extract solution twice a day for 2 hours after the morning and after dinner;

Group C1 and D1: Each dose is 100 mL, 2 times a day;

Group C2 and D2: 100 mL per dose, 2 times a day;

Group C3 and D3: Each dose is lOOmL, 2 times a day;

After 7 days of continuous administration, the number of bowel movements was counted by the user, and the presence or absence of a body sensation was observed and the change in facial skin gloss was observed 7 days earlier.

The effect of Tremella fuciformis extract on improving constipation and the improvement of skin gloss in constipation patients are shown in Table 3. The improvement effect of different concentrations of Tremella fuciformis polysaccharide extract on constipation is shown in Fig. 7; Tremella fuciformis extract is effective for stools with normal bowel movements. The laxative effect and the improvement of the skin gloss of the user are shown in Table 4. The lubricating effect of the different concentrations of the Tremella polysaccharide extract solution on constipation patients and normal stools is shown in Fig. 8. Tremella fuciformis extract improves constipation Group C1 C2 C3 Tremella polysaccharides 0.1% 0.2% 0.5% concentration of extract

Number of people 6 people 6 people 6 people constipation improvement situation invalid: 4 invalid: 1 invalid: 1 person (weekly improvement: 2 improvement: 3 improvement: 2 significant improvement: 0 significant improvement: 2 significant improvement : 3 people

Stool lubrication 17% 50% 100% Constipation aggravation None No No

Other discomfort

No change in facial skin gloss: No change in 3 people: No change in 2 people: 0 person observed slightly increased gloss: 2 people slightly increased gloss: 1 person slightly increased gloss: 2 Gloss increased significantly: 1 person's luster increased significantly: 3 people

Significant increase in luster: 4 people

Note: Invalid: The number of stools does not change

Improvement: Increased number of bowel movements 1~2 times

Table 4 Effect of Tremella polysaccharide extract on normal people's laxative

Group D1 D2 D3

Tremella polysaccharides 0.1% 0.2% 0.5%

Concentration of matter

Number 6 people 6 people 6 people

Weekly number of bowel movements unchanged: 5 people unchanged: 4 people unchanged: 4 people

Increase: 1 person increase: 2 people increase: 2 people Stool lubrication 50% 50% 100%

Cause constipation None No No

Other discomfort

No change in facial skin gloss: No change in 3 people: No change in 2 people: 1 person observed slightly increased gloss: 3 people slightly increased gloss: 3 slightly increased luster: 2 people's luster increased significantly: 0 people's luster increased significantly : 3 people's gloss increased significantly: 1

People

The experimental results show that: 1) Tremella fuciformis polysaccharide extract has the function of lubricating stool and can improve constipation;

2) The improvement effect increases significantly with the increase of the concentration of the polysaccharide extract of Tremella fuciformis;

3) The effect of gender and age on the improvement effect is not obvious;

4) There is no side effect on the body of constipation patients, and it will not cause bloating, abdominal pain and other feelings;

5) Tremella fuciformis extract has no side effects on normal people, no bloating, and the stool is relatively lubricated;

6) Tremella heteropolysaccharide extract has the effect of improving the skin's luster and improving the effect, and the effect of the increase is higher with the increase of the concentration of the tremella polysaccharide extract.

Summary: It can be seen from the above examples that Tremella fuciformis extract and Tremella fuciformis (WSK) are good promoters for promoting the growth and/or reproduction of probiotics, and also a good probiotic for promoting digestive or reproductive systems. A promoting factor for growth and/or reproduction of bacteria that regulates the balance of microbial flora in the large intestine. Moreover, the white fungus heteropolysaccharide extract and the white fungus heteropolysaccharide also have a laxative effect, which may be caused by promoting the growth and/or reproduction of probiotics in the digestive system, the white fungus heteropolysaccharide extract and the white fungus miscellaneous The polysaccharide can excrete toxins through the laxative and absorb the nutrients of the tremella polysaccharides to achieve the purpose of detoxification and beauty. Therefore, Tremella fuciformis extract and Tremella fuciformis (WSK) are healthy and nutritious foods.

Claims

Rights request

1. Use of a extract of Tremella fuciformis or Tremella heteropolysaccharide for the preparation of a medicament, a food or a health supplement for promoting the growth and/or reproduction of probiotics.

2. Use of a white fungus heteropolysaccharide or a tremella heteropolysaccharide extract for the preparation of a medicine, a food or drink or a health care product having a function of improving constipation.

3. Use of a white fungus heteropolysaccharide or a white fungus heteropolysaccharide extract for the preparation of a medicine, a food or a health care product having a detoxifying beauty function.

4. Use according to claim 1, characterized in that the probiotics are Bifidobacteria and/or Lactobacillus.

The use according to claim 4, characterized in that the Bifidobacterium is Bifidobacterium bifidum, and the Lactobacillus is a subspecies of Lactobacillus delbrueckii.

6. The use according to claim 5, characterized in that the Bifidobacterium bifidum is Bifidobacterium ATCC 2952, and the B. bulgaricus subsp. bulgaricus is Lactobacillus bulgaricus ATCC 11480c

The use according to any one of claims 1 to 6, wherein the medicine is a powder, a tablet, a capsule (including soft and hard capsules), a liquid, a suspension, an emulsion, an oral solution. , the jelly or the granule; the food or drink is a drink or food containing a concentrate and/or a dried product of the tremella or the fungus extract of the white fungus, and the use of the food or drink is not limited, and may be used in powders, tablets, Capsules (including soft and hard capsules), granules, granules, jelly, beverages, dairy products, yoghurt or snacks, etc.; the health care products are powders, tablets, capsules (including soft capsules and hard capsules) Capsules), liquids, suspensions, emulsions, oral liquids, jelly or granules.

8. Use of a Tremella heteropolysaccharide or a Tremella heteropolysaccharide extract for promoting the growth and/or reproduction of probiotics in vitro.

9. Use according to claim 8, characterized in that the probiotics are Bifidobacteria and/or Lactobacillus.

The use according to claim 9, characterized in that the Bifidobacterium is Bifidobacterium ATCC 2952, and the Lactobacillus is Lactobacillus bulgaricus ATCC 11480c