WO2018045946A1 - Produit de fermentation de poria cocos et son procédé de préparation - Google Patents

Produit de fermentation de poria cocos et son procédé de préparation Download PDF

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WO2018045946A1
WO2018045946A1 PCT/CN2017/100537 CN2017100537W WO2018045946A1 WO 2018045946 A1 WO2018045946 A1 WO 2018045946A1 CN 2017100537 W CN2017100537 W CN 2017100537W WO 2018045946 A1 WO2018045946 A1 WO 2018045946A1
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fermentation product
hydrazine
acid
fermentation
product according
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PCT/CN2017/100537
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Chinese (zh)
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陈科豪
陈思涵
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杏辉天力(杭州)药业有限公司
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Priority to CN201780055209.XA priority Critical patent/CN109689879A/zh
Publication of WO2018045946A1 publication Critical patent/WO2018045946A1/fr

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    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L31/00Edible extracts or preparations of fungi; Preparation or treatment thereof
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/06Fungi, e.g. yeasts
    • A61K36/07Basidiomycota, e.g. Cryptococcus
    • A61K36/076Poria
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00Antineoplastic agents
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P33/00Preparation of steroids

Definitions

  • the present invention relates to a fermentation product, particularly a hydrazine fermentation product obtained by using a solid substrate for fermentation culture of hydrazine.
  • the medicinal material refers to the dried sclerotium of the fungus fungus (Poria cocos (Schw.) Wolf).
  • the fungus is often parasitic on the roots of pine trees. It coats the roots of the trees and grows into a ball-like shape, resembling sweet potato.
  • the outer skin is light brown or dark brown (skin), and the interior is pink or white (clam meat).
  • belongs to aerobic fungi, mostly parasitic on pines such as Yunnan pine, red pine, Simao pine, masson pine, and Pinus taiwanensis.
  • Carbon sources such as cellulose and hemicellulose
  • nitrogen sources and minerals are obtained by decomposing pine roots. Wait for nutrients. The hyphae grow on the pine wood, and the fruit body can grow under certain conditions, and the spores can be released. The spores will regenerate into hyphae in a suitable environment, thereby repeating the cycle.
  • fermenting products containing high amounts of ruthenium triterpenoids can be provided by solid-state fermentation technology, by fermenting sputum with a solid substrate containing root crops and/or melon crops.
  • the method discovered by the inventor of the present invention has the advantages of environmental protection function, simple steps, easy maintenance of conditions, time saving, etc., and can provide a fermentation product containing a high amount of ruthenium triterpenes at a relatively low manufacturing cost, which is economically advantageous.
  • An object of the present invention is to provide a Poria cocos fermentation product in which the total amount of the following compounds (1) to (12) in the hydrazine fermentation product is not based on the total weight of the hydrazine fermentation product. Less than 0.15 wt%:
  • PA pachymic acid
  • DPA dehydropachymic acid
  • TTA Trametenolic acid
  • EA Eburicoic acid
  • the ratio of the total amount of the compounds (1) to (6) to the total amount of the compounds (7) to (12) is from 0.5 to 25; wherein, based on the total weight of the hydrazine fermentation product The total amount of the compounds (1) to (12) in the hydrazine fermentation product is not less than 0.3% by weight.
  • the hydrazine fermentation product of the present invention is a solid fermentation product of hydrazine, and the solid substrate used in the solid fermentation comprises a rhizome crop, a melon crop or a combination thereof.
  • the melon crop is a pumpkin
  • the root crop is at least one of the following: sweet potato, potato, taro and yam; particularly preferably, the root crop is sweet potato.
  • the hydrazine fermentation product of the present invention can be obtained by the following method:
  • the solid substrate comprises a rhizome crop, a melon crop or a combination of the foregoing.
  • the water content of the substrate may be adjusted to be 40% to 90% as needed before performing step (c).
  • the fermentation culture in the step (c) is carried out at a temperature of from 22 ° C to 32 ° C; more preferably, it is carried out at a temperature of from 22 ° C to 32 ° C and a relative humidity of from 30% to 70%;
  • the fermentation is carried out at a temperature of 22 ° C to 32 ° C and a relative humidity of 30% to 70% for more than 2 weeks.
  • the total amount of the compounds (1) to (12) in the hydrazine fermentation product is not less than 0.6% by weight based on the total weight of the hydrazine fermentation product, and The total amount of the compounds (7) to (12) in the hydrazine fermentation product is not less than 0.2% by weight; when the fermentation culture in the step (c) is carried out for 9 weeks or more, the hydrazine fermentation is based on the total weight of the hydrazine fermentation product.
  • the total amount of the compounds (7) to (12) in the product is not less than 0.4% by weight; when the step (c) The fermentation culture is carried out for 13 weeks or more, and the total amount of the compounds (7) to (12) in the hydrazine fermentation product is not less than 0.6% by weight based on the total weight of the hydrazine fermentation product.
  • solid state fermentation refers to a process of cultivating a fungus (for example, a sputum strain) using a solid substrate as a nutrient source; a "solid matrix” refers to a substrate containing a crop such as a rhizome and/or a melon.
  • the so-called “three meats of the meat department” refers to the extraction and purification of tannic acid, dehydroabietic acid, toluic acid, dehydromomoic acid, porcine acid C, 3-epi-dehydro-moic acid, etc.
  • Triterpenoids in the flesh the so-called “small scorpion” refers to dehydroabietic acid, ceric acid, ruthenium acid A, dehydroporosporin, ruthenium B, and porphyrin
  • the triterpenoids are extracted and purified from the phloem; the so-called “ ⁇ ” refers to the sum of the three mites of the scorpion and the scorpion scorpion; the so-called “matrix water content” refers to the matrix The total weight of the root-stem crops and/or melon crops and the pure water added, and the proportion of water.
  • has the functions of regulating the immune system, enhancing immunity, anti-allergy, anti-inflammatory, anti-tumor, lowering blood sugar, promoting nutrient absorption, etc.
  • the traditional saplings provided by the traditional pine section cultivation method It is only about 0.07 ⁇ 0.01% (based on the total weight of the medicinal herbs). Therefore, The industry continues to develop methods to effectively increase the content of ruthenium triterpenoids in medicinal herbs.
  • a patent document (such as TW I469741B) is known as a method of fermenting a culture medium in which a cereal or a bean is used as a sputum strain, and the patent document shows a product obtained by using wheat and brown rice as a fermentation substrate.
  • the crude triterpene content was 0.08%
  • the crude triterpene content of the product obtained by using coix seed as a fermentation substrate was 0.26%
  • the crude triterpene content of the product obtained by using oatmeal as a fermentation substrate was 0.35%.
  • cereals and legumes themselves contain crude triterpenoids. Therefore, the crude triterpenes shown in this patent document are not the total content of triterpenoids, but rather triterpenoids (or triterpenoids) and triterpenoids. The total content.
  • fermenting culture of alfalfa with a solid substrate containing root crops and/or melon crops can provide a high amount of triterpenoids (including triterpenoids and The sputum fermentation product of the scorpion scorpion), wherein the content of scorpion scorpion accounts for more than 60% of the total content of medlar.
  • the aforementioned mash fermentation product in addition to having the nutrient composition of cockroaches, can also retain the nutrient components of some rhizome crops and melon crops.
  • the present invention provides a hydrazine fermentation product, wherein the total amount of the following compounds (1) to (12) in the hydrazine fermentation product is not less than 0.15% by weight based on the total weight of the hydrazine fermentation product:
  • PA pachymic acid
  • DPA dehydropachymic acid
  • TTA Trametenolic acid
  • EA Eburicoic acid
  • the ratio of the total amount of the compounds (1) to (6) to the total amount of the compounds (7) to (12) is from 0.5 to 25;
  • the total amount of the compounds (1) to (12) in the hydrazine fermentation product is not less than 0.3% by weight based on the total weight.
  • the hydrazine fermentation product of the present invention may be a solid fermentation product of hydrazine, wherein the solid substrate used in the solid state fermentation contains a rhizome crop, a melon crop or a combination of the foregoing.
  • the melon crop is preferably a pumpkin
  • the root crop is preferably at least one of the following: sweet potato, potato, taro and yam, and especially using sweet potato.
  • the wild species of sweet potato originated in the tropical region of the Americas, and is characterized by strong resistance to pests and diseases and easy cultivation, and is currently widely planted all over the world.
  • the nutritional value of sweet potato is high. Every 100 grams of fresh sweet potato contains 28.6 grams of sugar, 1 kilogram of crude protein, 13 milligrams of vitamin C, 34 milligrams of calcium, as well as phosphorus, carotene and other nutrients.
  • Sweet potato skin is rich in polysaccharides, which can lower blood cholesterol and maintain blood vessel elasticity. For the prevention of cardiovascular diseases such as hardening of the arteries and hypertension.
  • Sweet potato is rich in dietary fiber, so eating sweet potato can get quite fullness and can also help gastrointestinal motility.
  • Sweet potato is also suitable as a nutrient source for the sputum strain.
  • the potato belongs to the perennial herb of the Solanaceae family. When cultivated artificially, it is usually used as an annual or two-year crop. Its underground tubers are edible. It is the third most important food crop in the world, and its annual output is second only to wheat and corn. The rice is close. Potato tubers have bud eyes and can be used for artificial planting. Potato has high nutritional value and medicinal value. It is rich in carbohydrates and can supply sufficient heat. It also contains protein, minerals (phosphorus, calcium, etc.), vitamins and other nutrients, and rich dietary fiber. And after eating it will produce considerable satiety. Potato skin is rich in chlorogenic acid and lipoic acid. Chlorogenic acid has anti-oxidation and anti-cancer effects, while lipoic acid can prevent blemishes and prevent skin aging. Potatoes also contain other dietary antioxidants that help slow down aging and prevent disease.
  • Taro is a perennial herb of the genus Araceae, which is produced all year round. It is usually used as an annual crop during artificial cultivation. Its underground tubers are edible and can be used as medicine. The islands of Oceania are traditional staple foods of some indigenous people. Each section of the taro has a axillary bud that can be used for artificial planting, and it has the strongest axillary bud activity at the lower and middle nodes. A large amount of small granular starch is stored in the steamed bread, and the cooked food has good digestibility. It also contains protein and rich minerals (calcium, phosphorus, potassium, magnesium, fluorine, etc.) and vitamins (B group, C). It is generally believed that taro can promote immune function and digestive function, so it is often used in the dietary planning of the elderly and patients.
  • Dioscorea is a perennial grass-wound vine of the family Dioscorea, whose roots are called yam.
  • Yam is a high-sugar, high-protein, low-fat food, rich in a variety of minerals and vitamins needed by the human body. Appropriate consumption can increase satiety and promote gastrointestinal motility. Yam also has many effects such as anti-oxidation, lowering blood sugar, lowering blood pressure, improving blood lipids, regulating female hormones, etc. Appropriate consumption is beneficial to human health.
  • the pumpkin belongs to the annual climbing herb of the cucurbitaceae, and the starch is rich as a staple food, and contains an antioxidant component such as ⁇ -carotene, vitamin C and E, and a lutein. , phenol, selenium, mannitol and other ingredients that have anti-cancer and anti-cancer effects.
  • Pumpkin has a high zinc content and helps prevent prostate enlargement or disease, so it is listed as one of 30 anti-cancer fruits and vegetables by the US Food and Drug Administration (FDA).
  • FDA US Food and Drug Administration
  • studies have shown that eating pumpkin can provide effects such as activating the immune system, regulating blood sugar, protecting the cardiovascular system, and promoting skin health.
  • the present invention also provides a method of preparing the hydrazine fermentation product of the present invention comprising the steps of:
  • the solid substrate comprises a rhizome crop, a melon crop or a combination thereof;
  • the melon crop is preferably a pumpkin, and the root crop is preferably at least one of the following: sweet potato, potato, taro and yam, especially Sweet potato is better.
  • the sputum strain may be subjected to a pre-activation treatment as needed before the step (b).
  • the sputum strain is inoculated on the medium plate with an inoculum of 5 mm in diameter, and the medium plate inoculated with the sputum strain is placed at a temperature of 27 ⁇ 5° C., relative humidity.
  • the preactivated alfalfa strain was inoculated onto the solid substrate after about 6 days of pre-activation of the alfalfa strain under 40 ⁇ 10%.
  • Examples of the aforementioned pre-activated medium plates for the sputum strain include, but are not limited to, a malt extract agar (MEA) plate, a potato dextrose agar (PDA) plate.
  • the water content of the substrate before the step (c), and adjust the ambient temperature and humidity during the step (c), and the step (c).
  • the length of time For example, the water content of the fermentation substrate can be adjusted to 40% to 90% before the step (c), and the steps can be carried out at a temperature of 22 ° C to 32 ° C and a relative humidity of 30% to 70%.
  • the fermentation substrate has a water content adjusted to 75% to 85% before fermentation, followed by fermentation at a temperature of 27 ⁇ 5 ° C and a relative humidity of 40 ⁇ 10%. Cultivate for more than 2 weeks.
  • the total amount of the compounds (1) to (12) in the hydrazine fermentation product is not less than 0.6% based on the total weight of the obtained hydrazine fermentation product.
  • the total amount of the compounds (7) to (12) in the hydrazine fermentation product is not less than 0.2% by weight; when the fermentation culture in the step (c) is carried out for 9 weeks or more, the total weight of the hydrazine fermentation product obtained is
  • the total amount of the compounds (7) to (12) in the hydrazine fermentation product is not less than 0.4% by weight; when the fermentation culture in the step (c) is carried out for 13 weeks or more, the total weight of the hydrazine fermentation product obtained is The total amount of the compounds (7) to (12) in the hydrazine fermentation product is not less than 0.6% by weight.
  • the content of strontium triterpenoids is significantly higher than that of the traditional sorghum cultivation method (the scorpion medicinal material is only about 0.07 ⁇ 0.01%), and can be directly used for making food,
  • the feed or the addition of both of the above may be provided by further extraction and purification treatment.
  • the mash fermentation product of the present invention may be sterilized, dried, and/or pulverized as needed, and then prepared into, for example, dairy products, processed meat products, breads, noodles, biscuits, and ingots. , capsules, juices, teas, sports drinks, nutritious drinks, animal feeds, etc., or the addition of these products, but not limited to this.
  • the sputum strain (Poria cocos; deposited in the Food Industry Development Research Institute of the consortium, registered strain No. BCRC39126) was inoculated into the malt extract agar with an inoculum of 5 mm (mm) diameter.
  • MEA malt extract purchased from Medical Microbiology Technology Co., Ltd., product number: 218630; Yangcai purchased from Medical Microbiology Technology Co., Ltd., product number: LP0011
  • the MEA plate inoculated with the sputum strain was placed at the temperature At 27 ⁇ 5 ° C and a relative humidity of 40 ⁇ 10%, pre-activation of the sputum strain was carried out for about 6 days.
  • the preactivation of the aforementioned sputum strain can also be carried out by replacing the MEA plate with a plate of potato dextrose agar (PDA; purchased from Medical Microbiology Technology Co., Ltd., product number: 254920).
  • B-1 Take fresh sweet potato, potato, taro, yam and pumpkin, wash and remove the outer skin, cut into small pieces with a particle size of less than 1 cm and place in a fermentation container (such as airtight bottle, fermenter). . Then, pure water was added to each fermentation vessel in the following weight ratio, and then sterilized at 121 ° C for 30 minutes, and after cooling, the first, second, third, fourth, and fifth solid substrates were used for subsequent experiments:
  • B-2 Take fresh sweet potato, cut the outer skin and cut into sweet potato with a particle size of less than 1 cm, then dry the sweet potato diced to constant weight with 50 °C hot air and place it in the fermentation container (such as airtight bottle and fermenter). )in. Next, pure water was added to the fermentation vessel at a weight ratio of 1:1.5 (dried sweet potato:water), and then sterilized at 121 ° C for 30 minutes, and after cooling, it was a solid group of the sixth group (water content) 60%) for use in subsequent experiments.
  • B-3 Take the fresh sweet potato, wash and remove the outer skin and cut it into the potato chips with the particle size less than 1 cm, then dry the sweet potato diced to constant weight with 50 °C hot air and place it in the fermentation container (such as airtight bottle). , fermenter). Next, pure water is added to the fermentation vessel at a weight ratio of 1:1.5 (de-peeled sweet potato:water), and then sterilized at 121 ° C for 30 minutes, and after cooling, it is a solid group of the seventh group (including The amount of water is 60%) for use in subsequent experiments.
  • Example 1 Solid state fermentation culture of alfalfa strain
  • Example 2 ⁇ 3 ⁇ content of ⁇ solid-state fermentation product
  • step 2 2. performing alcohol extraction on the powder obtained in step 1 by ultrasonic vibration, repeating the foregoing extraction step for a total of three times;
  • the crude extract obtained in the step 3 is concentrated under reduced pressure at 60 ° C to remove the solvent to obtain an extract;
  • Ultra-high performance liquid chromatography was used to analyze the content of triterpenoids or triterpenoids in the extract obtained in step 4, and the results are shown in Table 1. Table 3 (shown as a percentage).
  • Table 1, Table 2, and Table 3 show the analytical knots of the ruthenium fermentation products of Examples 1-1, 1-2, and 1-3, respectively. fruit.
  • the content of triterpenoids in the meat portion is 100% based on the total weight of the extract, and the calculation of tannic acid, dehydroabietic acid, toluic acid, dehydromomoic acid, porcine acid C, and 3-epi-dehydrogenation
  • the total content of tortonic acid in the extract; the content of triterpenoids in the molting is 100% based on the total weight of the extract, and the calculation of dehydroabietic acid, ceric acid, ruthenium acid A, dehydroporospores
  • the total content of acid, ruthenium acid B and porphyric acid in the extract; the content of ruthenium osmium is the sum of the content of scorpion and the content of scorpion.
  • the fermentation of cockroaches using the solids of Groups 1 to 7 can provide strontium fermentation containing high amounts of strontium triterpenoids.
  • the product, especially the mash fermentation product provided by the fermentation of the first, sixth and seventh groups of solid substrates, has a significantly higher ruthenium content.
  • the content of strontium triterpenoids is significantly higher than that of the traditional sorghum cultivation method (the ⁇ 3 ⁇ content is only about 0.07 ⁇ 0.01%). It can be used directly to make food, feed or additives of the foregoing, or can be provided by further extraction and purification treatment.

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Abstract

La présente invention concerne un produit de fermentation de Poria cocos. Par rapport au poids total du produit de fermentation de Poria cocos, la proportion totale des composés (1) à (12) suivants est supérieure ou égale à 0,15 % en poids : (1) l'acide pachymique (PA), (2) l'acide déhydropachymique (DPA), (3) l'acide tumulosique (TA), (4) l'acide déhydrotumulosique (DTA), (5) l'acide polyporénique C (PAC), (6) l'acide 3-épi-déhydrotumulosique (EDTA), (7) l'acide déhydrotraméténolique (DTTA), (8) l'acide traméténolique (TTA), (9) l'acide poricoïque A (PAA), (10) l'acide déhydroéburicoïque (DEA), (11) l'acide poricoïque B (PAB), et (12) l'acide éburicoïque (EA). Le produit de fermentation de Poria cocos est préparé en faisant appel au procédé suivant consistant à : (a) placer un substrat solide dans un récipient de fermentation ; (b) inoculer une souche de Poria cocos sur le substrat solide ; (c) cultiver par fermentation, le substrat solide contenant des cultures rhizomateuses, des cultures de melon ou une combinaison de celles-ci.
PCT/CN2017/100537 2016-09-06 2017-09-05 Produit de fermentation de poria cocos et son procédé de préparation WO2018045946A1 (fr)

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CN110988191A (zh) * 2019-12-23 2020-04-10 哈尔滨市康隆药业有限责任公司 一种参术儿康糖浆的hplc含量测定方法

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