WO2007030975A1 - Procédé de réalisation de synchronisation de commandes dans la prise en charge d'un système de fichiers de mémoire non volatile à traitement multifils - Google Patents
Procédé de réalisation de synchronisation de commandes dans la prise en charge d'un système de fichiers de mémoire non volatile à traitement multifils Download PDFInfo
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- WO2007030975A1 WO2007030975A1 PCT/CN2005/001574 CN2005001574W WO2007030975A1 WO 2007030975 A1 WO2007030975 A1 WO 2007030975A1 CN 2005001574 W CN2005001574 W CN 2005001574W WO 2007030975 A1 WO2007030975 A1 WO 2007030975A1
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- tremella
- heteropolysaccharide
- extract
- lactobacillus
- food
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/10—Laxatives
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/16—Emollients or protectives, e.g. against radiation
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/02—Antidotes
Definitions
- 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.
- 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.
- 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..
- 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.
- 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.
- 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 ⁇ - ( 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.
- 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 ⁇ -(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.
- 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 -
- the structural unit has a molecular weight of 2578.5 or 2724.7.
- the sugar structure is composed of ⁇ -(1-3) mannose (Manp) as the main chain skeleton, and two ⁇ -(1-2) xylose (Xylp) and three ⁇ -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 ⁇ ,
- 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.
- Tremella heteropolysaccharide extract of the present invention comprises the following steps -
- 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.
- the purification was carried out by the Sevag method and the dialysis method.
- 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.
- 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.
- 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.
- 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.
- Tremella fuciformis extract and Tremella fuciformis 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.
- 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;
- FIG. 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.
- Tremella heteropolysaccharide extract and Tremella heteropolysaccharide 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
- 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 ⁇ m; 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 ⁇ freeze-dried, and pulverized to obtain a white fungus heteropolysaccharide extract.
- Product yield 0.55% (based on the weight of the fermentation broth).
- 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).
- Tremella heteropolysaccharide promotes the growth and/or reproduction of bifidobacteria
- Test strain Bifidobacteria ATCC 2952, purchased from the American Type Culture Collection
- 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)
- Detection method Liquid quantitative limiting dilution method.
- 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.
- 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.
- 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
- Test strain Lactobacillus ATCC 11480, purchased from the Institute of Microbiology, Chinese Academy of Sciences. Medium: MRS Broth Medium
- 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)
- 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.
- anaerobic bacteria basal medium Shanghai Institute of Biological Products
- Detection method Liquid quantitative limiting dilution method.
- 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.
- the Tremella polysaccharide 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.
- 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.
- 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;
- Tremella heteropolysaccharide (WSK) obtained in Example 4;
- Bottle (PE bottle) cleaning method 75% alcohol soaking, clean room 24 hours to dry;
- 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;
- Tremella heteropolysaccharide has no side effects on normal people, no bloating, and the stool is relatively lubricated;
- 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.
- Tremella Heteropolysaccharide Extract Solution Sterile treatment, ensuring the health and safety of the user, and eliminating the influence of bacteria on the test results;
- the Tremella heteropolysaccharide extract is the Tremella fuciformis extract (WSK-1) obtained in Example 1, The purity of Tremella fuciformis polysaccharide is 80%;
- Bottle (PE bottle) cleaning method 75% alcohol soaking, clean room 24 hours to dry;
- 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;
- Tremella fuciformis extract improves constipation Group C1 C2 C3 Tremella polysaccharides 0.1% 0.2% 0.5% concentration of extract
- Tremella fuciformis polysaccharide extract has the function of lubricating stool and can improve constipation;
- Tremella fuciformis extract has no side effects on normal people, no bloating, and the stool is relatively lubricated;
- 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.
- Tremella fuciformis extract and Tremella fuciformis 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.
- 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.
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Description
银耳杂多糖或其提取物的新用途 技术领域
本发明涉及一种杂多糖的用途; 尤其涉及一种银耳杂多糖或其提取物在促进益生菌 生长和 /或繁殖方面的用途, 以及在改善便秘功能和排毒养颜美容功能等方面的用途。 技术背景
双歧杆菌是人体内的生理性细菌, 革兰氏阳性、 无芽孢、 不运动、 严格厌氧, 在人 体肠道内占绝对优势, 终生存在于人体, 与人的健康息息相关, 是一种对人体具有许多 重要生理功能的有益菌。 随着微生物学的迅猛发展, 双歧杆菌的临床保健作用逐渐被人 们所认识和接受。 人的生理、 免疫、 营养、 消化、 抗肿瘤、 抗衰老、 药物效能等都离不 开人体自身携带的微生物群, 而人体肠道内的双歧杆菌则具有有病治病, 无病防老和保 健的作用。 例如, 双歧杆菌能形成生理屏障, 阻止致病菌的定植和入侵, 保护宿主免受 致病菌侵害, 能改善蛋白质代谢, 提供多种 B种维生素, 促进钙铁吸收, 提高免疫力, 增强抗肿瘤能力等多种作用。 若人体内的双歧杆菌的不足意味着人体发生健康危机, 则 需要及时补充。
乳酸杆菌也是人体内重要的的生理性细菌, 革兰氏阳性、 无芽孢、 不运动、 严格厌 氧, 具有多种生理功能。 乳酸杆菌能维持肠道及生殖道菌群的平衡, 在清除某些致癌剂 或预防致癌等环节上发挥关键作用。 乳酸杆菌的减少可能导致肠道菌群失调, 病原菌乘 机大量繁殖, 其代谢产物中的某些细菌酶能催化致癌前体物质向致癌物质转化。 而乳酸 杆菌可恢复肠道菌群平衡, 抗肠道中的病原菌生长, 抑制细菌酶的产生及降低其活性, 还可结合致癌物质随粪便排出, 减少了其形成、 活化及滞留机会。
益生元是指一类非消化性的物质, 但可作为底物被肠道正常菌群利用, 能够选择性 地刺激肠内一种或几种有益菌生长和繁殖, 抑制有害细菌生长, 对宿主健康具有有益的 作用, 如水苏精、 低聚果糖(包括异麦芽低聚糖、 半乳糖低聚糖、 果糖低聚糖等。 因这 些物质可以促进双歧杆菌生长和 /或繁殖, 故也称为 "双歧因子"。 低聚果糖是国际公认 的双歧因子, 但它们都是低聚糖类, 分子量比较小。 通过服用益生元类物质可以促进体
内益生菌生长和 /或繁殖, 进一步可以改善人体消化吸收***功能, 有助于改善便秘和促 进人体新陈代谢, 而体内毒素被及时排出, 又可以达到美容禾 g养颜的效果。
银耳 (Tremella faciformis Berk ) , 又称白木耳、 桑鹅、 《清异录》。 属银耳目 (Tremellales), 银耳科(Tremellacease), 银耳属 (Tremella)。 具有滋阴润肺、 益气和血 之功效。 银耳杂多糖是从银耳子实体 (或其微生物发酵液) 中提取分离的一种酸性杂多 糖。 中国自古以来将银耳称之为高级补品和药品, 药食两用之佳品是宫廷上等进补食品。
随着科学的不断发展以及技术手段的不断提升, 人们对银耳有了重新的认识, 国内 外对银耳的主要成分——银耳杂多糖进行了大量的研究, 表明银耳杂多糖具有提高免疫 力、 抗肿瘤、 抗衰老、 降血糖、 血脂、 促进蛋白质核酸合成、 抗血栓等药理作用。 日本 已经把它广泛地运用于保健品、 化妆品、 药品中。 银耳的开发和应用, 引起了科学界的 极大兴趣, 受到国际医药界和化妆品领域的广泛重视, 成为当今研究开发的热门话题。
但是至今为止, 银耳杂多糖的组成和结构以及对应的功能却很少有报导。 本申请人 运用特殊分离纯化工艺将银耳中活性多糖加以提炼, 经现代分析技术检测鉴定, 该银耳 杂多糖的结构以 α - ( 1-3 )甘露糖为主链,主要由重量百分比为 70-93%的中性总糖、 6-28% 的葡萄糖醛酸、 0.1-3%的结合蛋白组成;该银耳杂多糖的平均分子量为 85-160万道尔顿, 其中分子量大于 6000道尔顿的占 90% (重量) 以上, 具体参见本申请人于 2005年 1月 18曰申请的申请号为 200510023427.4的专利申请, 并将其商品名命名为 WSK; 该专利 申请还公开了银耳杂多糖及其提取物具有保湿滑润皮肤和粘膜、 增强润滑感、 抗氧化衰 老、 抗过敏、 增稠稳定等功能。 但是到目前为止, 还没有银耳杂多糖具有促进肠道及消 化***益生菌的增殖和改善便秘功能以及排毒养颜美容功能和用途的相关报道。 发明内容
本发明的目的是为了提供一种申请号为 200510023427.4 的专利申请中的商品名为 WSK的银耳杂多糖或银耳杂多糖提取物在制备促进益生菌生长和 /或繁殖的药品、饮食品 或保健品中的用途。
本发明的另一目的是提供一种申请号为 200510023427.4 的专利申请中的商品名为 WSK的银耳杂多糖或银耳杂多糖提取物在制备具有改善便秘功能的药品、饮食品或保健
品中的用途。
本发明的又一目的是提供一种申请号为 200510023427.4 的 利申请中的商品名为 WSK的银耳杂多糖或银耳杂多糖提取物在制备具有排毒养颜美容功能的药品、饮食品或 保健品中的用途。
本发明所述的银耳杂多糖的结构以 α- (1-3) 甘露糖为主链, 主要由重量百分比为 70-93%的中性总糖、 6-28%的葡萄糖醛酸、 0.1-3%的结合蛋白组成; 该银耳杂多糖的平均 分子量为 85-160万道尔顿, 其中分子量大于 6000道尔顿的占 90% (重量) 以上。其中, 所述的中性总糖主要包括下列含量的糖基: 35-60%甘露糖、 10-25%岩藻糖、 8-18%木糖 和 5-15%葡萄糖, 上述含量是以银耳杂多糖重量计。
该银耳杂多糖的侧链有多种不同的变化组合,其中的一种组合已通过 X光衍射方法 测定为如下结构式 I所示的结构单元-
→3) - a -Manp- (1—3) - α - Manp- (1→3) -α-Manp- (1→3) -a - Manp- (1—3) - (接下行)
2 2 2
t † t
1 1 1
β -Xylp β -GlcAp β -GlcAp
(接上行)
α - (1→3)
( I )
该结构单元的分子量为 2578.5或 2724.7。 该糖结构是以 α- (1-3)甘露糖(Manp) 为主 链骨架, 其 2位碳原子上连有 2个 β- (1-2)木糖(Xylp), 3个 β-葡萄糖醛酸(GlcAp), 1或 2个岩藻糖(Fuc), 1个甘露糖为侧链。
本发明所述的银耳杂多糖提取物主要包括重量百分比为 80-98%的上述银耳杂多糖, 0.1-3.3%的游离蛋白及 0.5-8%的灰分; 该银耳杂多糖提取物的平均分子量为 75-140万道 尔顿, 其中分子量大于 6000道尔顿的占 75-98% (重量)。
本发明所述的银耳杂多糖提取物可以采用下列制备方法得到-
1 )将银耳子实体原料用水在 80-100°C下搅拌提取得粗提液^ ,
2)将步骤 1 )所得的粗提液进行过滤, 滤液最终通过孔径为 0.1-1 μ ηι的过滤器;
3 )在步骤 2)所得的滤液中加入乙醇, 使乙醇的最终体积浓度为 50-85%, 收集沉 淀;
4)将步骤 3)所得的沉淀干燥。
其中, 该银耳子实体原料为干燥的银耳子实体原料, 将该干燥的银耳子实体原料用 水浸泡、 膨胀并清洗干净 (包括去黄褐色基部), 每次用 20-100倍、 多次提取时不超过 200倍 (重量) 于该干燥的银耳子实体原料量的水在 80-100°C下搅拌提取得粗提液。
该银耳子实体原料也可以为新鲜银耳子实体原料、 保鲜冷藏的新鲜银耳子实体原料 或冷冻的新鲜银耳子实体原料, 将该银耳子实体原料去黄褐色基部并清洗干净, 每次用 2-15倍、多次提取时不超过 30倍(重量)于该银耳子实体原料量的水在 80-100°C下搅拌 提取得粗提液。
本发明所述的银耳杂多糖提取物的另一种制备方法包括如下步骤-
1 )将按照常规微生物发酵方法 (参考文献①银耳深层发酵条件的研究, 吴大康等; 食品科学; 2002, Vol.23, No山 64; ②银耳菌分离方法的研究, 张炳炽; 中国食用菌; Vol.13, No.3; 22)获得的银耳发酵液加热至 80-100°C、保温 0.5-2小时, 过滤, 清液最终 通过孔径为 0.1-1 μ m的过滤器得滤液;
2)在步骤 1 )所得的滤液中加入乙醇, 使乙醇最终体积浓度为 50-85%, 收集沉淀;
3)将步骤 2)所得的沉淀进行干燥。
本发明所述的银耳杂多糖的制备方法包括将采用上述制备方法制得的提取物进一步 纯化的步骤。
其中, 该纯化采用 Sevag法及透析法来实现。 当然, 如本领域技术人员所知, 其它 的多糖纯化方法, 如色谱分离法等也能制得本发明银耳杂多糖。
所述的益生菌可为各种益生菌,主要为双歧杆菌和 /或乳酸杆菌;较佳地为消化*** (如肠道) 中的双歧杆菌和 /或乳酸杆菌, 或生殖*** (如生殖道) 中的乳酸杆菌。
所述的双歧杆菌 Bifidobaterinm为不规则形的专性厌氧菌, 有 32个种, 优选两歧双 歧杆菌、 青春双歧杆菌或婴儿双歧杆菌, 更优选两歧双歧杆菌; 所述的两歧双歧杆菌较
佳地为双歧杆菌 ATCC 2952。 所述的乳酸杆菌 Lactobacillus为革兰氏阳性无芽孢杆菌, 有 56个种, 优选德氏乳杆菌、 干酪乳杆菌和明串株乳杆菌, 更优选德氏乳杆菌; 所述的 德氏乳杆菌较佳地为德氏乳杆菌保加利亚亚种, 更佳地为保加利亚乳杆菌 ATCC 11480。
所述的药品可以为现有技术中的各种剂型, 如粉剂、 片剂、 胶囊剂 (包括软胶囊和 硬胶囊)、液体剂、 悬浮液、 乳剂、 口服液、果冻或冲剂; 所述的饮食品包括饮品和食品, 为含有银耳杂多糖或银耳杂多糖提取物的浓缩液和 /或干燥物的饮品或食品, 对饮食品的 应用没有限制, 可以为现有技术中的各种剂型, 如可以使用在粉剂、 片剂、 胶囊剂 (包 括软胶囊和硬胶囊)、 颗粒、 冲剂、 果冻、 饮料、 乳制品、 酸奶或小点心烘烤品等; 所述 的保健品可以为现有技术中的各种剂型, 如粉剂、片剂、胶囊剂(包括软胶囊和硬胶囊)、 液体剂、 悬浮液、 乳剂、 口服液、 果冻或冲剂等。
根据本发明, 所述的药品、 饮食品或保健品中还可以含有其他促进益生菌生长和 / 或繁殖的益生元。
所述的饮食品或保健品中还可以含有其他各种可食用的营养成分, 以起到更佳的营 养保健的作用。
本发明所述的银耳杂多糖可以是粗提过的含有杂质的银耳杂多糖粗品, 也可以是银 耳杂多糖纯品。
本发明的再一目的是提供一种申请号为 200510023427.4 的专利申请中的商品名为 WSK 的银耳杂多糖或银耳杂多糖提取物在体外促进益生菌生长和 /或繁殖的促进剂中的 用途。
在本发明中, 所述的促进剂的作用或功能类似于双歧因子, 它们的不同之处在于: 双歧因子是促进双歧杆菌生长和 /或繁殖的物质,而促进剂是促进益生菌(包括双歧杆菌) 生长和 /或繁殖的物质。 本发明所述的促进剂添加在药品、 饮食品和保健品中, 服用后可 以促进体内益生菌的生长和繁殖; 对含有益生菌的药品、 饮食品和保健品中添加本发明 所述的促进剂可以增强该类产品的功效。 本发明所述的促进剂在体外, 例如可以用于培 养基中, 以培养益生菌, 使得以更快捷的方式培养出更多的有益菌, 从而降低成本。
所述的益生菌可为各种益生菌, 主要为双歧杆菌和 /或乳酸杆菌。
所述的双歧杆菌优选两歧双歧杆菌 B.bifidum,更优选双歧杆菌 ATCC 2952;所述的
乳酸杆菌优选德氏乳杆菌, 更优选德氏乳杆菌保加利亚亚种 L.delbraeckii subsp.bulgaricus, 最优选保加利亚乳杆菌 ATCC 11480。
本发明的积极进步效果在于: 银耳杂多糖提取物及银耳杂多糖(WSK)对益生菌的 生长和 /或繁殖具有促进作用,如直接调节人体肠道菌、增殖体内双歧杆菌和 /或乳酸杆菌, 将具有较广的发展前景。而且银耳杂多糖对益生菌如双歧杆菌和乳酸杆菌的促增殖作用, 将可作为促进剂 (类似于双歧因子的功能或作用)进一步开发研制, 并促进微生态学的 发展。 本发明的含有银耳杂多糖或银耳杂多糖提取物的药品、 饮食品或保健品经口服后 具有改善便秘的功能和排毒养颜美容的功能, 其通过排出体内毒素以及吸收银耳杂多糖 中的营养物质可以起到美容养颜的作用。 附图说明
图 1为无糖、低聚糖和银耳杂多糖对双歧杆菌 ATCC 2952生长和繁殖影响的比较图; 图 2A为对照组 1的双歧杆菌 ATCC 2952生长和繁殖的照片;
图 2B为实验组(银耳杂多糖组) 的双歧杆菌 ATCC 2952生长和繁殖的照片; 图 3为无糖、 低聚糖和银耳杂多糖对保加利亚乳杆菌 ATCC 11480生长和繁殖影响 的比较图;
图 4A为对照组 1的保加利亚乳杆菌 ATCC 11480生长和繁殖的照片;
图 4B为实验组(银耳杂多糖组)的保加利亚乳杆菌 ATCC 11480生长和繁殖的照片; 图 5为不同浓度的银耳杂多糖对便秘的改善效果的示意图;
图 6为不同浓度银耳杂多糖溶液对便秘患者及正常人大便的润滑作用的示意图。 图 7为不同浓度的银耳杂多糖提取物 (WSK-1 )对便秘的改善效果的示意图; 图 8为不同浓度银耳杂多糖提取物(WSK-1 ) 溶液对便秘患者及正常人大便的润滑 作用的示意图。 具体实施方式
现根据本申请人的申请号为 200510023427.4的专利申请中的制备方法制备所需的银 耳杂多糖提取物及银耳杂多糖 (WSK), 将其详细描述如下:
实施例 1
将干燥的银耳子实体用水浸泡, 膨胀后, 去其黄褐色基部并清洗干净; 把洗净后的 原料放入提取罐中, 加 100倍于干料的水, 100Ό水温下搅拌提取 4h, 然后过滤,清液最 终通过孔径为 O.l-ΐμιη的过滤器; 均匀搅拌下, 将滤清液和乙醇充分混合, 使混合液体 中的乙醇最终浓度为 85%, 收集沉淀, 用纯乙醇脱水, 收集沉淀; -40°C冷冻千燥, 粉碎 得银耳杂多糖提取物 (WSK-1 )。 产品收率 20% (以干银耳重量计), 银耳杂多糖的纯度 为 80%。
实施例 2
将保鲜冷藏的银耳, 去其黄褐色基部并清洗干净; 把洗净后的原料放入提取罐中, 加 15倍于鲜料的水, 10CTC水温下搅拌提取 4h,然后过滤,清液最终通过孔径为 0.1-1μιη 的过滤器; 滤清液加热浓缩, 得浓缩液; 均匀搅拌下, 将滤清液和乙醇充分混合, 使混 合液体中的乙醇最终浓度为 85%, 收集沉淀, 用纯乙醇脱水脱色, 收集沉淀; 80°C热风 干燥, 粉碎得银耳杂多糖提取物。 产品收率 2.1% (以鲜银耳重量计)。
实施例 3
将微生物发酵获得的银耳发酵液, 加温 100°C, 0.5h, 常规过滤, 弃滤渣, 清液最终 通过孔径为 0.1-1μηι的过滤器; 均匀搅拌下, 将滤清液和乙醇充分混合, 使混合液体中 的乙醇最终浓度为 85%, 收集沉淀, 用 100%乙醇脱水脱色, 收集沉淀; -40Ό冷冻干燥, 粉碎得银耳杂多糖提取物。 产品收率 0.55% (以发酵液重量计)。
实施例 4
将实施例 1〜3银耳杂多糖提取物用 600倍 (重量 )去离子水溶解,溶解后,加入 200mL 的氯仿: 正丁醇 =4: 1 (体积比) 的混合溶液, 充分混合后, 用离心机离心 (4000r/min, 30分钟), 收集上清液。 将离心后的上清液与上述氯仿正丁醇混合溶液按 3: 1 (体积比) 混合, 并用离心机离心 (4000r/min, 30分钟), 收集上清液。 重复以上操作, 反复处理 至水相在 280nm处的 OD值(使用 752紫外光栅分光光度计, 上海精密科学仪器厂) 不 再下降为止。 水相减压浓缩至三分之一, 浓缩液与水相 3倍量 (体积比) 的 95%乙醇充 分混合, 4000r/min, 30分钟离心, 得沉淀物。沉淀物用 100倍(重量)去离子水溶解后, 离心, 去不溶物, 将清液装入 30cm长的透析袋内 (透析袋可以通过 5000分子量以下物
质)进行透析(两头扎好吊装入有去离子水的瓶内, 并使用去离子水流水冲洗 12小时), 收集 5000分子量以上物质。 经透析后的清液与 3 倍量 (体积) 纯乙 充分混合, 再次 4000r/min离心 30分钟, 得沉淀物, 沉淀物用无水乙醇脱水后, 60°C真空干燥, 获得纯 银耳杂多糖 (WSK)。
效果实施例 1 银耳杂多糖 (WSK) 促进双歧杆菌生长和 /或繁殖的作用
样品名称: 实施例 4得到的银耳杂多糖(WSK), 低聚果糖(无锡轻工大学食品学院 提供的标准品)
测试菌株:双歧杆菌(Bifidobacteria) ATCC 2952,购于美国菌种保存中心(American Type Culture Collection)
培养基: MRS肉汤培养基
灭菌方法: 在 90°C下, 灭菌 30分钟
实验处理分为以下三项- 对照组 1 : 培养基 6.3mL+菌液 0.3mL
对照组 2: 培养基 6.0mL+低聚果糖 0.3mL+菌液 0.3mL (该培养基中低聚果糖: 培' 养基的质量比为 1 : 80)
实验组: 培养基 6.0mL+银耳杂多糖样液 0.3mL+菌液 0.3mL (该培养基中银耳杂多: 糖: 培养基的质量比为 1 : 80)
实验方法: 液体培养法
请给出培养双歧杆菌的步骤: 用厌 02菌基础培养基(上海生物制品研究所)添加维 生素 K至 0.1wt%, 制成液体培养基, 在常规孵育箱 37Ό培养, 静止培养 48小时。
检测方法: 液体定量有限稀释法。
注:液体培养法的细菌定量计数 560nmOD值时: 1.0OD值 =6X 108个菌 /mL, 2.0OD 值 = 18 X 108个菌 /mL, 3.0OD值 =24X 108个菌 /mL。
实验结果表明: 在液体培养基中添加银耳杂多糖 (1 : 80稀释), 可以促进双岐杆菌 的生长和繁殖, 而且效果显著。 从图 1中可见, 相对于对照组 1 中无糖的培养基来说, 银耳杂多糖可显著地提高发酵液中双岐杆菌的数量, 银耳杂多糖对双歧杆菌的生长和繁 殖的促进作用甚至超过目前市场上常用作益生元的低聚果糖。而且,双歧杆菌 ATCC 2952
在 MRS肉汤培养基中的培养时间从 48小时提前到 24小时; 且经涂片染色可见, 与对照 组 1 比, 实验组的菌体增大粗壮, 在同一视野下菌量的密度明显地增 ¾, 见图 2A和图
2Bo 「
效果实施例 2 银耳杂多糖 (WSK) 促进乳酸杆菌生长和 /或繁殖的作用
样品名称: 实施例 4得到的银耳杂多糖(WSK), 低聚果糖(无锡轻工大学食品学院 提供的标准品)
测试菌株:保加利亚乳杆菌(Lactobacillus) ATCC 11480,购于中科院微生物研究所。 培养基: MRS肉汤培养基
培养基灭菌方法: 在 9CTC下, 灭菌 30分钟
实验处理分为以下三项:
对照组 1 : 培养基 6.3mL+菌液 0.3mL
对照组 2: 培养基 6.0mL+低聚果糖 0.3mL+菌液 0.3mL (该培养基中低聚果糖: 培 养基的质量比为 1: 80)
实验组: 培养基 6.0mL+WSK样液 0.3mL+菌液 0.3mL (该培养基中银耳杂多糖: 培' 养基的质量比为 1 : 80)
实验方法: 液体培养法
乳酸杆菌的培养: 用厌 02菌基础培养基(上海生物制品研究所)添加维生素 K至浓 度为 0.1wt%, 制成液体培养基, 在常规孵育箱 37°C培养, 静止培养 48小时。
检测方法: 液体定量有限稀释法。
注:液体培养法的细菌定量计数 560nmOD值时: 1.0OD值 =6X 108个菌 /mL, 2.0OD 值 = 18 X 108个菌 /mL, 3.0OD值 = 24 X 108个菌 /mL。
实验结果表明: 在液体培养基中添加银耳杂多糖 (1 : 80稀释), 可以促进乳酸杆菌 的生长和繁殖, 而且效果显著。 从图 3中可见, 相对于对照组 1中无糖的培养基来说, 银耳杂多糖可显著地提髙发酵液中乳酸杆菌的数量, 银耳杂多糖对乳酸杆菌的生长和繁 殖的促进作用甚至超过目前市场上常用作益生元的低聚果糖。 而且, 保加利亚乳杆菌 ATCC 11480在 MRS肉汤培养基中的培养时间从 48小时提前到 24小时; 且经涂片染色 可见, 与对照组 1 比, 实验组的菌体增大粗壮, 在同一视野下菌量的密度明显地增加,
见图 4A和图 4B。
效果实施例 3 测试银耳杂多糖(WSK)对便秘患者的便秘改善效果(是否有改善) 和对正常人的口服效果以及服用后脸部皮肤光泽的变化情况 (是否具有美容养颜的功 能)。
实验对象: 16名便秘患者,其症状是排便次数 3次 /周或大便干燥,性别年龄不限, 分成 3组, Al、 A2、 A3组, 分别服用 0.1wt%、 0.2wt%、 0.5wt%的银耳杂多糖溶液, 观 察大便状况和脸部皮肤光泽的变化情况; 16名排便正常者, 分成 3组, Bl、 B2、 B3组, 分别服用 0.1wt%、 0.2wt%、 0.5wtQ/。的银耳杂多糖溶液, 观察大便状况和脸部皮肤光泽的 变化情况。
银耳杂多糖溶液的配制说明: 无菌处理, 保证饮用者的健康安全, 排除细菌对试验 结果的影响;
1 )配制足量的 0.1wt%、 0.2wt%、 0.5wt%银耳杂多糖溶液供试验用, 该银耳杂多糖. 是实施例 4得到的银耳杂多糖 (WSK);
2)配制时选用细菌检测结果 <20cfu/g无菌的银耳杂多糖粉末, 防腐剂选用山梨酸鉀. (添加量 0.1〜0.2wt%,参考中国食品添加剂手册, 山梨酸钾说明书及微生物试验结果);
3 )配制方法: 取少量蒸馏水加热煮沸 10分钟, 冷却至 70〜80°C后加入山梨酸钾, 溶解备用, 剩余蒸馏水加入银耳杂多糖粉末均质机搅拌充分溶解, 稍后将山梨酸钾水溶 液加入, 混合均勾, 整个操作过程严格无菌;
4)瓶子 (PE瓶)清洗方法: 75%酒精浸泡, 洁净室 24小时晾干;
5)要求: 无菌、 干燥、 密封, 用于罐装配制好的银耳杂多糖溶液。
实验方法: 便秘患者和排便正常者在观察期内, 于每天早、 晚餐之后 2小时服用银 耳杂多糖溶液, 每天 2次;
A1和 B1组: 每次服用量为 100mL, 每天 2次;
A2和 B2组: 每次服用量为 lOOmL, 每天 2次;
A3和 B3组: 每次服用量为 100mL, 每天 2次;
连续服用 7天后, 由服用者对排便次数进行统计,并记录有无身体异样感以及对比 7 天前观察脸部皮肤光泽的变化情况。
银耳杂多糖改善便秘的效果和便秘患者的皮肤光泽改善情况见表 不同浓度的银耳 杂多糖对便秘的改善效果见图 5;银耳杂多糖对大便正常者的润肠通便效果和服用者的皮 肤光泽改善情况见表 2,不同浓度银耳杂多糖溶液对便秘患者及正常人大便的润滑作用见 图 6。
表 1 银耳杂多糖改善便秘的效果
注: 无效: 大便次数不变
改善: 排便次数增加 1〜2次
明显改善: 排便次数增加 2〜4次, 且感觉大便明显润滑
表 2 银耳杂多糖对正常人润肠通便的效果
组别 B1 B2 B3 银耳杂多糖的 0.1% 0.2% " 0.5%
浓度
人数 5人 6人 5人
周排便次数 不变: 4人 不变: 5人 不变: 4人
增加: 1人 增加: 1人 增加: 1人 大便润滑 60% 50% 100%
导致便秘 无 无 无
其他不适感 无 无 无
脸部皮肤光泽 无变化: 2人 无变化: 2人 无变化: 1人
观察 光泽略有增加: 3人 光泽略有增加: 2 光泽略有增加: 1人 光泽明显增加: 0人 人 光泽明显增加: 3人 光泽明显增加: 2
人
实验结果表明: 1 )银耳杂多糖具有润滑大便的作用, 并能改善便秘;
2) 改善效果随银耳杂多糖浓度增加明显增加;
3 )性别、 年龄对改善效果的影响不明显;
4)对便秘患者的身体无副作用, 不会导致胃胀、 腹痛等感觉;
5 )银耳杂多糖对正常人没有副作用, 无腹胀感, 大便比较润滑;
6)银耳杂多糖具有改善皮肤光泽的养颜美容作用, 改善效果随银耳杂多糖浓度增加 明显增加。
效果实施例 4 测试银耳杂多糖提取物对便秘患者的便秘改善效果(是否有改善)和 对正常人的口服效果以及服用后脸部皮肤光泽的变化情况(是否具有美容养颜的功能)。
实验对象: 18名便秘患者,其症状是排便次数 3次 /周或大便干燥,性别年龄不限, 分成 3组, Cl、 C2、 C3组, 分别服用 0.1wt%、 0.2wt%、 0.5wt%的银耳杂多糖提取物溶 液, 观察大便状况和脸部皮肤光泽的变化情况; 18名排便正常者, 分成 3组, Dl、 D2、 D3组, 分别服用 0.1wt%、 0.2wt%、 0.5wt%的银耳杂多糖提取物溶液, 观察大便状况和
脸部皮肤光泽的变化情况。
银耳杂多糖提取物溶液的配制说明: 无菌处理, 保证^:用者的健康安全, 排除细菌 对试验结果的影响;
1 )配制足量的 0.1wt%、 0.2wt%、 0.5wt%银耳杂多糖提取物溶液供试验用, 该银耳 杂多糖提取物是实施例 1得到的银耳杂多糖提取物(WSK-1 ),银耳杂多糖的纯度为 80%;
2)配制时选用细菌检测结果 <20db/g无菌的银耳杂多糖提取物粉末, 防腐剂选用山 梨酸钾(添加量 0.1〜0.2wt%, 参考中国食品添加剂手册, 山梨酸钾说明书及微生物试验 结果);
3 )配制方法: 取少量蒸镏水加热煮沸 10分钟, 冷却至 70〜80°C后加入山梨酸钾, 溶解备用, 剩余蒸馏水加入银耳杂多糖提取物粉末均质机搅拌充分溶解, 稍后将山梨酸 钾水溶液加入, 混合均匀, 整个操作过程严格无菌;
4)瓶子 (PE瓶)清洗方法: 75%酒精浸泡, 洁净室 24小时晾干;
5)要求: 无菌、 干燥、 密封, 用于罐装配制好的银耳杂多糖提取物溶液。
实验方法: 便秘患者和排便正常者在观察期内, 于每天早、 晚餐之后 2小时服用银 耳杂多糖提取物溶液, 每天 2次;
C1和 D1组: 每次服用量为 lOOmL, 每天 2次;
C2和 D2组: 每次服用量为 100mL, 每天 2次;
C3和 D3组: 每次服用量为 lOOmL, 每天 2次;
连续服用 7天后, 由服用者对排便次数进行统计,并记录有无身体异样感以及对比 7 天前观察脸部皮肤光泽的变化情况。
银耳杂多糖提取物改善便秘的效果和便秘患者的皮肤光泽改善情况见表 3,不同浓度 的银耳杂多糖提取物对便秘的改善效果见图 7;银耳杂多糖提取物对大便正常者的润肠通 便效果和服用者的皮肤光泽改善情况见表 4,不同浓度银耳杂多糖提取物溶液对便秘患者 及正常人大便的润滑作用见图 8。 银耳杂多糖提取物改善便秘的效果
组别 C1 C2 C3 银耳杂多糖提 0.1% 0.2% 0.5% 取物的浓度
人数 6人 6人 6人 便秘改善情况 无效: 4人 无效: 1人 无效: 1人 (周排便次 改善: 2人 改善: 3人 改善: 2人 明显改善: 0人 明显改善: 2人 明显改善: 3 人
大便润滑 17% 50% 100% 便秘加重 无 无 无
其他不适感 无 无 无
脸部皮肤光泽 无变化: 3人 无变化: 2人 无变化: 0人 观察 光泽略有增加: 2人 光泽略有增加: 1人 光泽略有增加: 2 光泽明显增加: 1人 光泽明显增加: 3人 人
光泽明显增加: 4 人
注: 无效: 大便次数不变
改善: 排便次数增加 1〜2次
明显改善: 排便次数增加 2〜4次, 且感觉大便明显润滑
表 4 银耳杂多糖提取物对正常人润肠通便的效果
组别 D1 D2 D3
银耳杂多糖提 0.1% 0.2% 0.5%
取物的浓度
人数 6人 6人 6人
周排便次数 不变: 5人 不变: 4人 不变: 4人
增加: 1人 增加: 2人 增加: 2人
大便润滑 50% 50% 100%
导致便秘 无 无 无
其他不适感 无 无 无
脸部皮肤光泽 无变化: 3人 无变化: 2人 无变化: 1人 观察 光泽略有增加: 3人 光泽略有增加: 3 光泽略有增加: 2人 光泽明显增加: 0人 人 光泽明显增加: 3人 光泽明显增加: 1
人
实验结果表明: 1 ) 银耳杂多糖提取物具有润滑大便的作用, 并能改善便秘;
2) 改善效果随银耳杂多糖提取物浓度增加明显增加;
3) 性别、 年龄对改善效果的影响不明显;
4) 对便秘患者的身体无副作用, 不会导致胃胀、 腹痛等感觉;
5) 银耳杂多糖提取物对正常人没有副作用, 无腹胀感, 大便比较润滑;
6)银耳杂多糖提取物具有改善皮肤光泽的养颜美容作用, 改善效果随银耳杂多糖提 取物浓度增加明显增加。
总结: 从上述实施例可知, 银耳杂多糖提取物及银耳杂多糖 (WSK) 是一种良好的 促进益生菌生长和 /或繁殖的促进剂, 也是一种良好的促进消化***或生殖***中益生菌 生长和 /或繁殖的促进因子, 其可以调节大肠中微生物菌群平衡。 而且该银耳杂多糖提取 物及银耳杂多糖还具有润肠通便的作用, 该作用可能是由于促进消化***中益生菌的生 长和 /或繁殖而引起的, 该银耳杂多糖提取物及银耳杂多糖通过润肠通便将毒素排出体外 以及吸收银耳杂多糖的营养成分可以达到排毒养颜美容的目的。 所以, 银耳杂多糖提取 物及银耳杂多糖 (WSK) 是一种健康营养的食品。
Claims
1、 一种银耳杂多糖或银耳杂多糖提取物在制备促进益生菌生长和 /或繁殖的药品、 饮食品或保健品中的用途。
2、一种银耳杂多糖或银耳杂多糖提取物在制备具有改善便秘功能的药品、饮食品或 保健品中的用途。
3、一种银耳杂多糖或银耳杂多糖提取物在制备具有排毒养颜美容功能的药品、饮食 品或保健品中的用途。
4、 根据权利要求 1 所述的用途, 其特征在于所述的益生菌为双歧杆菌和 /或乳酸杆 菌。
5、 根据权利要求 4所述的用途, 其特征在于所述的双歧杆菌为两歧双歧杆菌, 所述 的乳酸杆菌为德氏乳杆菌保加利亚亚种。
6、 根据权利要求 5所述的用途, 其特征在于所述的两歧双歧杆菌为双歧杆菌 ATCC 2952, 所述的德氏乳杆菌保加利亚亚种为保加利亚乳杆菌 ATCC 11480c
7、 根据权利要求 1〜6任一项所述的用途, 其特征在于所述的药品为粉剂、 片剂、 胶囊剂 (包括软胶囊和硬胶囊)、 液体剂、 悬浮液、 乳剂、 口服液、 果冻或冲剂; 所述的 饮食品为含有银耳杂多糖或银耳杂多糖提取物的浓缩液和 /或干燥物的饮品或食品, 对饮 食品的应用没有限制, 可以使用在粉剂、 片剂、 胶囊剂 (包括软胶囊和硬胶囊)、 颗粒、 冲剂、 果冻、 饮料、 乳制品、 酸奶或小点心烘烤品等; 所述的保健品为粉剂、 片剂、 胶 囊剂 (包括软胶囊和硬胶囊)、 液体剂、 悬浮液、 乳剂、 口服液、 果冻或冲剂。
8、 一种银耳杂多糖或银耳杂多糖提取物在体外促进益生菌生长和 /或繁殖的促进剂 中的用途。
9、 根据权利要求 8 所述的用途, 其特征在于所述的益生菌为双歧杆菌和 /或乳酸杆 菌。
10、根据权利要求 9所述的用途,其特征在于所述的双歧杆菌为双歧杆菌 ATCC 2952, 所述的乳酸杆菌为保加利亚乳杆菌 ATCC 11480c
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US8435541B2 (en) | 2010-09-02 | 2013-05-07 | Bath & Body Works Brand Management, Inc. | Topical compositions for inhibiting matrix metalloproteases and providing antioxidative activities |
CN115005442A (zh) * | 2022-06-15 | 2022-09-06 | 中国科学院新疆理化技术研究所 | 一种天山雪莲多糖提取物制备方法与应用 |
CN115299537A (zh) * | 2022-07-05 | 2022-11-08 | 北京吉澳医学科技有限责任公司 | 一种具有美容养颜、改善便秘的饮品及其制备方法和应用 |
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CN101579117B (zh) * | 2009-03-26 | 2013-06-19 | 大山合集团有限公司 | 一种银耳软胶囊健康食品 |
CN102370004A (zh) * | 2010-08-24 | 2012-03-14 | 上海辉文生物技术有限公司 | 一种银耳杂多糖或其提取物在液体乳制品中的应用 |
CN102336840B (zh) * | 2011-10-17 | 2012-11-21 | 浙江大学 | 一种三股螺旋银耳多糖及其制备方法和应用 |
TW201345537A (zh) * | 2012-05-10 | 2013-11-16 | Univ Asia | 保護視網膜細胞之銀耳多醣及其製造方法 |
JP2017195830A (ja) * | 2016-04-28 | 2017-11-02 | 日本食菌工業株式会社 | 脂肪蓄積予防用の食品組成物 |
JP6969067B2 (ja) * | 2016-05-10 | 2021-11-24 | 丸善製薬株式会社 | 抗炎症剤、抗老化剤および美白剤 |
JP6670166B2 (ja) * | 2016-05-10 | 2020-03-18 | 丸善製薬株式会社 | 化粧料 |
CN105949337A (zh) * | 2016-05-13 | 2016-09-21 | 福建农林大学 | 一种促双歧杆菌增殖的鲜银耳多糖提取方法 |
CN107019657A (zh) * | 2016-12-14 | 2017-08-08 | 上海辉文生物技术股份有限公司 | 一种美白保湿组合物及其制备方法和使用方法 |
CN110590968B (zh) * | 2019-09-05 | 2021-09-28 | 浙江工业大学 | 一种银耳低聚糖的制备方法及其应用 |
JP7253283B2 (ja) * | 2020-02-27 | 2023-04-06 | 丸善製薬株式会社 | 化粧料および飲食品組成物 |
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Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
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US8435541B2 (en) | 2010-09-02 | 2013-05-07 | Bath & Body Works Brand Management, Inc. | Topical compositions for inhibiting matrix metalloproteases and providing antioxidative activities |
CN115005442A (zh) * | 2022-06-15 | 2022-09-06 | 中国科学院新疆理化技术研究所 | 一种天山雪莲多糖提取物制备方法与应用 |
CN115005442B (zh) * | 2022-06-15 | 2023-07-25 | 中国科学院新疆理化技术研究所 | 一种天山雪莲多糖提取物制备方法与应用 |
CN115299537A (zh) * | 2022-07-05 | 2022-11-08 | 北京吉澳医学科技有限责任公司 | 一种具有美容养颜、改善便秘的饮品及其制备方法和应用 |
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CN1931879A (zh) | 2007-03-21 |
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