CN113730476A - Paecilomyces hepiali strain fermentation composition and preparation method and application thereof - Google Patents
Paecilomyces hepiali strain fermentation composition and preparation method and application thereof Download PDFInfo
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- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
- A61K31/197—Carboxylic acids, e.g. valproic acid having an amino group the amino and the carboxyl groups being attached to the same acyclic carbon chain, e.g. gamma-aminobutyric acid [GABA], beta-alanine, epsilon-aminocaproic acid, pantothenic acid
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- A61P5/00—Drugs for disorders of the endocrine system
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- A61P5/00—Drugs for disorders of the endocrine system
- A61P5/24—Drugs for disorders of the endocrine system of the sex hormones
- A61P5/30—Oestrogens
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- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
Abstract
The invention belongs to the technical field of application of paecilomyces hepiali strain, and discloses a paecilomyces hepiali strain fermentation composition, and a preparation method and application thereof. The desugarization of beta-glucosidase in a culture solution obtained after fermentation of paecilomyces hepiali strain is utilized to convert SDG in the linseed lignan into SECO, the SECO is easier to absorb, and the bioavailability of the linseed lignan is improved from 15% to 75%; under the action of beta-glucosidase, a plurality of glycosides or glycosides in raw materials of the red rose with double petals, the gamma-aminobutyric acid, the red ginseng, the red dates, the medlar and the cranberry can be converted into aglycone which is easy to be absorbed by a human body, and the bioavailability of active ingredients in the raw materials is remarkably improved. The formula is used for enriching blood and activating blood, tonifying qi and nourishing liver, improving metabolism capability, promoting microcirculation in a body, accelerating internal circulation, and conditioning female endocrine dyscrasia so as to improve the syndrome caused by female estrogen reduction.
Description
Technical Field
The invention belongs to the technical field of application of paecilomyces hepiali strain, and particularly relates to a paecilomyces hepiali strain fermentation composition, and a preparation method and application thereof.
Background
The endocrine system and the nervous system regulate the metabolism and physiological functions of the human body together, wherein the endocrine system participates in regulating a plurality of physiological activities of the human body such as metabolism, growth and development, reproductive aging and the like by secreting various hormones, and cooperates with various biological enzymes to maintain the relative stability of the internal environment of the human body so as to adapt to complicated and variable internal and external changes of the human body. When the endocrine system of a human body is disordered, corresponding clinical manifestations can be caused, and especially women have special physiological processes of menstruation, leukorrhea, abortion, obstetrics and the like, and bear double pressure of work and families in daily life, so that endocrine dyscrasia is more likely to occur, and various gynecological diseases such as irregular menstruation, dysmenorrhea, amenorrhea, hyperplasia of mammary glands and the like are caused.
According to the traditional Chinese medicine, endocrine dyscrasia is the manifestation of yin deficiency and is caused by qi and blood stasis, and the qi and blood stasis can be caused by blood stasis retention in vivo, vein obstruction, external toxin invasion, postpartum lochiorrhea and the like, so that qi and blood are smooth, essence and blood are nourished to the whole body, blood circulation is promoted, and comprehensive conditioning is performed from inside to outside.
The existing gynecological medicine only has single function due to the limitation of the formula, and the existing gynecological medicine for treating gynecology is matched with chemical medicines with few side effects, although the effect is quick, the relapse condition in a short time is very common. The pure Chinese medicine prescription is used for the best treatment, but the curative effect course of the pure Chinese medicine gynecological medicine is very long, and the curative effect cannot be cured radically; and the traditional Chinese medicine decocting process is complex, the time is limited, and the high-efficiency production cannot be realized.
Paecilomyces hepiali, a Chinese medicine institute of Chinese academy of sciences in China, separated from larvae (stiff insects) of Cordyceps hepiali Hepialus armoricanus Oberthiri in 1982 to obtain more than 10 strains, and culturing and morphologically researching 4 strains of the strains, and identifying and naming the strains as Paecilomyces hepiali. The Cs-4 strain is separated from fresh Cordyceps collected from Qinghai Hualong by the institute of medicine of Chinese academy of sciences in 1982, and is identified as Paecilomyces hepiali. Paecilomyces hepiali Chen & Dai (Paecilomyces hepiali Chen & Dai) mycelium is obtained by separating fresh Cordyceps from Paecilomyces hepiali Chen & Dai (Paecilomyces hepiali Chen & Dai) by artificial submerged fermentation culture of Paecilomyces hepiali Bain fungus belonging to Paecilomyces of Moniliaceae, and has been widely used as substitute of Cordyceps in Chinese patent medicine and health food.
The paecilomyces hepiali fermentation and Chinese herbal medicine combined prescription is used, so that the drug effects of various Chinese herbal medicines are promoted to achieve a more sufficient prescription, and the contents pursued by a plurality of researchers are formed.
Disclosure of Invention
In order to solve the problems in the prior art, the invention aims to provide a paecilomyces hepiali strain fermentation composition, and a preparation method and application thereof.
The technical scheme adopted by the invention is as follows: a paecilomyces hepiali strain fermentation composition is mainly prepared from the following raw materials in parts by mass:
20-60 parts of linseed concentrated powder, 10-30 parts of rose powder with double petals, 10-30 parts of gamma-aminobutyric acid, 10-55 parts of red ginseng powder, 10-55 parts of red date powder, 10-55 parts of medlar powder, 10-55 parts of cranberry powder and 20-100 parts of paecilomyces hepiali powder.
Flax seed concentrated powder, also known as flax (Linum usittissimum L), belongs to the genus Linum of the family Linaceae and is one of the oldest crops. Flaxseed is the seed of flax, with a hard outer shell, in which Flaxseed Lignans (Flaxseed Lignans) are mainly present. Flax seed lignans are also known as phytoestrogens because they are very similar in their chemical structure to human estrogens. The linseed lignan has bidirectional regulation effect on estrogen level in a human body: A. estroidogenic effects: because of its chemical structure closely resembling human estrogen, flax seed lignan SDG is converted to Enterolactone (END) and Enterodiol (ENL) after ingestion of SDG when estrogen levels are low in the body. END and ENL are structurally similar to estrogens, with antioxidant activity and weak estrogenic effects. Can balance the action of estrogen in vivo by connecting with the receptor, and can relieve climacteric symptoms to a certain extent. B. Inhibition of estrogenic effects: when the estrogen level in the body is deceptively high, END and ENL can also be used as antiestrogen molecules, because the structure is very similar to the main form of estrogen, but the same estrogen action is not provided, and after being combined with some breast cell receptors with active proliferation, the estrogen action is blocked and the growth of the cells is inhibited. Chemical composition of linseed lignan: flax seed lignans are natural products produced by the metabolism of phenylpropanoids. The highest content in flaxseed is Secoisolariciresinol DiGlucoside (SDG). The aglycone of the diglycoside SDG is Secoisolariciresinol (SECO). SDG is also not present in flaxseed in free form, but part of the SDG is covalently bound to 3-hydroxy-3-methyl-glutaric acid (HMGA) via an ester bond, and on average 4 HMGA molecules are cross-linked with each other per 5 SDG molecules, resulting in an oligomer with a relative molecular mass of about 4000. Metabolites of linseed lignans: flax seed lignans are converted to various metabolites including Enterolactone (ENL) and Enterodiol (ED)) and their phase II metabolites, such as glucuronic acid conjugates. The conversion of diisohydroxyresinol diglucoside (SDG) of lignans of flaxseed can be divided into four catalytic reactions, in the order O-deglycosylation (conversion of SDG to its aglycone, SECO), O-demethylation (conversion of SECO to 2, 3-bis (3' -hydroxybenzyl) butyrolactone/2, 3-bis (3, 4-dihydroxybenzyl) butane-1, 4-diol), dehydrogenation (conversion of 2, 3-bis (3, 4-dihydroxybenzyl) butane-1, 4-diol to ED) and dihydroxylation (conversion of ED to ENL).
The rose flower is a representative of traditional Chinese roses, is large and bright in color, strong in fragrance, thick in multiple petals, 8 cm in single flower diameter and 6 g in single flower weight, and becomes a main variety for production and cultivation. The rose with double petals is the only edible rose for I. The traditional Chinese medicine considers that the rose is sweet and slightly bitter in taste and warm in nature, and has the most obvious effects of regulating qi, resolving depression, promoting blood circulation, removing blood stasis, regulating menstruation and relieving pain. In addition, the rose is very mild in drug property, can warm and nourish heart, liver and blood vessels of people, soothes depression in the body, and has the effects of calming, soothing and resisting depression. Women often have emotional irritability before or during menstruation, and rose can be drunk for regulating effect. Nowadays, the pressure of work and life is getting more and more, even not menstrual period, can also drink some roses more, pacify, stabilize mood. The flos Rosae Rugosae is rich in vitamin A, C, B, E, K and tannin, and can improve endocrine disturbance, and is helpful for relieving fatigue and promoting wound healing. Regulating qi and blood, regulating female physiological problems, promoting blood circulation, caring skin, regulating menstruation, promoting urination, relieving gastrointestinal nerve, preventing wrinkle, preventing frostbite, and caring skin. It is also suitable to take some of the above-mentioned herbs to massage when the body is tired and sore. The rose buds are made into dry flowers, 5 to 7 flowers are used each time, a small bunch of tender and sharp green tea is added, three red dates (needing to be denucleated) are added, the tea is brewed with boiled water every day for drinking, heart fire can be removed, spirit is kept full, vitality of people is increased, and the rose buds can be white and red in appearance and keep youthful and beautiful after being drunk for a long time. Conditioning the body: the rose tea is mild in property, can alleviate emotion, balance endocrine, enrich the blood, beautify and protect the skin, has the effect of conditioning the liver and the stomach, can eliminate fatigue and improve physique, is fragrant and elegant in taste, can alleviate emotion, relieve depression, improve endocrine dyscrasia, relieve waist soreness and back pain, eliminate fatigue and wound healing, moisten and beautify the skin, promote blood circulation, protect the liver, harmonize the stomach and nourish the liver, eliminate fatigue and promote blood circulation. Can be used for treating chronic gastritis and hepatitis. Soothing liver-qi stagnation, regulating qi and blood: soothing liver and relieving depression, strengthening spleen and lowering fire, dissipating blood stasis and the like, can treat abdominal psychroalgia, stomach cold accumulation, blood and qi smoothening, tranquilization, relaxing bowels, lowering fire and qi, regulating blood and qi, promoting blood circulation to remove blood stasis, relieving emotion, regulating endocrine, regulating qi and blood, and regulating female physiological problems; maintaining beauty and keeping young: most importantly, the tea has the effects of beautifying and beautifying, can remove black spots on skin, enables the skin to be tender, white and natural, and is helpful for preventing wrinkles when drunk frequently. Can improve constitution, and can be taken more for weight reduction due to stagnation of liver-qi, which is helpful for weight reduction; has the effects of enlarging breast and regulating menstruation; can also be used as a good product for relaxing bowel, maintaining beauty and keeping young and losing weight.
The physiological activity of the gamma-aminobutyric acid is mainly shown in the functions of tranquilizing nerves, resisting anxiety, reducing blood pressure, reducing blood ammonia, improving brain activity, promoting ethanol metabolism, preventing skin aging, preventing arteriosclerosis, efficiently losing weight and the like.
Red ginseng, nutrient components: the main component of Ginseng radix Rubri is triterpene saponin, and also contains volatile oil, polysaccharide, amino acids, microelements, polypeptide, etc. Triterpene saponin and polysaccharide of Ginseng radix Rubri are the main pharmacologically active components. The efficacy is as follows: the red ginseng is a cooked product of ginseng, has the effects of invigorating primordial qi, tonifying spleen and lung, promoting the production of body fluid and soothing the nerves, is milder in medicine property compared with the ginseng, and is used for treating deficiency of qi and blood, unhealthy hands and feet, dizziness and lassitude, limb weakness, fatigue, shortness of breath and dyspnea, frequent cold in stomach, long-term diarrhea, insomnia and dreaminess, impotence and frequent micturition and the like.
The red dates are warm in nature and sweet in taste, and contain protein, fat, sugar, calcium, phosphorus, iron and magnesium, abundant vitamin A, vitamin C, vitamin B1 and vitamin B2, and carotene and the like. Nourishing spleen and stomach, invigorating qi and spleen, nourishing blood and tranquillizing, promoting biochemical circulation of qi and blood, invigorating spleen and stomach, benefiting qi, nourishing blood and tranquillizing, delaying aging, and stimulating appetite.
Medlar, and the nutrient components: the medlar extract comprises the following main components: the molecular weight of Lycium barbarum polysaccharides (Lycium barbarum polysaccharides) is 22-25 kD, and the Lycium barbarum polysaccharides (Lycium barbarum polysaccharides) comprise 6 monosaccharide components including arabinose, glucose, galactose, mannose, xylose and rhamnose. The medlar also contains abundant carotene, vitamin A, vitamin C, vitamin E, calcium, iron, magnesium, phosphorus and carbohydrate. The efficacy is as follows: has the main effects of enhancing immunity, regulating immunity, promoting hemopoiesis, reducing blood lipid, preventing fatty liver, resisting tumor, and resisting aging. Has effects in replenishing vital essence, improving eyesight, and nourishing liver and kidney, and is suitable for people with asthenia, soreness of waist and knees, visual function deterioration, giddiness, and tinnitus.
Cranberry, nutrient content: the cranberry contains various flavonol compounds such as procyanidine, ellagic acid, phenolic acid, resveratrol, hyperoside, quercetin, myricetin, etc. The efficacy is as follows: the cranberry has the most unique effect of effectively preventing Urinary Tract Infection (UTI), inhibiting helicobacter pylori infection and having good effect of preventing gastric ulcer, gastric cancer and duodenal ulcer. Can inhibit oxidation of low density lipoprotein-cholesterol, reduce triglyceride level in blood, and prevent cardiovascular and cerebrovascular diseases such as atherosclerosis.
Preferably, the composition is mainly prepared from the following raw materials in parts by mass:
30-50 parts of linseed concentrated powder, 15-25 parts of rose powder with double petals, 15-25 parts of gamma-aminobutyric acid, 22-45 parts of red ginseng powder, 22-45 parts of red date powder, 22-45 parts of medlar powder, 22-45 parts of cranberry powder and 44-80 parts of paecilomyces hepiali powder.
Preferably, the composition is mainly prepared from the following raw materials in parts by mass:
40 parts of linseed concentrated powder, 20 parts of rose powder with double petals, 20 parts of gamma-aminobutyric acid, 30 parts of red ginseng powder, 30 parts of red date powder, 30 parts of medlar powder, 30 parts of cranberry powder and 60 parts of paecilomyces hepiali powder.
A preparation method of a paecilomyces hepiali strain fermentation composition comprises the following steps:
s1, selecting paecilomyces hepiali powder according to a corresponding proportion, adding water for dispersion, performing strain culture, and performing strain fermentation culture to obtain a paecilomyces hepiali fermentation culture solution;
s2, selecting linseed concentrated powder, double-petal red rose pollen, gamma-aminobutyric acid, red ginseng powder, red date powder, medlar powder and cranberry powder according to a corresponding proportion, and adding water for dispersing to respectively obtain linseed dispersion liquid, double-petal red rose dispersion liquid, gamma-aminobutyric acid dispersion liquid, red ginseng dispersion liquid, red jujube dispersion liquid, medlar dispersion liquid and cranberry dispersion liquid;
s3, mixing the paecilomyces hepiali fermentation culture solution, the linseed dispersion solution, the double red rose dispersion solution, the gamma-aminobutyric acid dispersion solution, the red ginseng dispersion solution, the red date dispersion solution, the medlar dispersion solution and the cranberry dispersion solution, uniformly mixing, standing for reaction, centrifuging and drying to obtain a finished product.
Preferably, in the step S1, when the paecilomyces hepiali powder is inoculated after being dispersed in water, the inoculation amount is 2.5-5 wt% of the paecilomyces hepiali powder in water dispersion liquid;
the strain culture comprises strain slant culture and strain shake flask culture;
the strain slant culture process comprises the following steps: the composition of slant culture medium includes 2-4 wt% of glucose, 0.2-0.4 wt% of peptone, 0.4-0.8 wt% of bran, 0.1-0.3 wt% of potassium dihydrogen phosphate, 0.03-0.05 wt% of magnesium sulfate, 0.1-0.3 wt% of histidine, 2-4 wt% of agar, pH: 5.0 to 7.5; and (3) sterilizing a culture medium: 0.1MPa for 30-40 min; inoculating the strain to a slant culture medium, and culturing at 15-20 deg.C for 7-9 days;
the strain shake flask culture process is as follows: the shake flask culture medium comprises 1-5 wt% of glucose, 0.2-0.4 wt% of peptone, 0.4-0.6 wt% of bran, 0.1-0.3 wt% of monopotassium phosphate, 0.03-0.05 wt% of magnesium sulfate, 0.03-0.05 wt% of zinc sulfate, 0.1-0.3 wt% of histidine, 10.05-0.1 wt% of vitamin B, and pH: 5.0 to 7.5; inoculating cultured strain into shake flask culture medium under aseptic condition, and culturing in 15-20 deg.C constant temperature shaking table under illumination for 7-9 days; the oscillating frequency of the reciprocating shaking table is 135 times/minute, and red light with the wavelength of 622-760 nm is used for irradiation in the whole shaking culture process.
Preferably, in the step S1, the strain fermentation culture includes a seed tank culture, a propagation tank culture and a fermentation tank culture;
the culture medium of the seeding tank comprises the following components: 1-2 wt% of hot-pressed soybean cake powder, 1-2 wt% of glucose, 1-2 wt% of sucrose, 0.1-0.2 wt% of monopotassium phosphate, 0.05 wt% of magnesium sulfate, 0.05 wt% of zinc sulfate, 0.1 wt% of histidine, 0.02 wt% of vitamin B1, 0.1 wt% of soybean oil, 0.05 wt% of adenine, pH: 5.0 to 7.5; under the aseptic condition, inoculating the strain cultured in the shake flask into a seed tank by adopting a pressure difference method according to the inoculation amount of 3 percent, wherein the temperature is 16 ℃, the tank pressure is 0.02MPa, and the ventilation volume is 1: standing and culturing for 4-5 days under the condition of 0.5 (v/v.min);
the propagation tank culture medium comprises the following components: 1-2 wt% of hot-pressed soybean cake powder, 1-2 wt% of glucose, 1-2 wt% of sucrose, 0.1 wt% of monopotassium phosphate, 0.04 wt% of magnesium sulfate, 0.03 wt% of zinc sulfate, 0.1 wt% of histidine, 0.1 wt% of vitamin B1, 0.2 wt% of soybean oil, 0.05 wt% of adenine, 0.05 wt% of uracil, pH: 5.0 to 7.5; under the aseptic condition, inoculating the cultured strains in the seed tank into a propagation tank by adopting a differential pressure method according to 9 percent of inoculation amount; at the temperature of 16 ℃, the tank pressure of 0.02MPa and the air flow of 1: standing and culturing for 4-5 days under the condition of 0.15 (v/v.min);
the culture medium of the fermentation tank: the composition of the fermenter medium was: 1-3 wt% hot-pressed soybean cake powder, 1 wt% glucose, 1 wt% sucrose, 0.2 wt% monopotassium phosphate, 0.04 wt% magnesium sulfate, 0.04 wt% zinc sulfate, 0.2 wt% histidine, 0.2 wt% vitamin B1, 0.2 wt% soybean oil, 0.05 wt% adenine, 0.05 wt% uracil, pH: 5.0 to 7.5; under the aseptic condition, inoculating the cultured strains in the propagation tank into a fermentation tank by adopting a differential pressure method according to the inoculation amount of 5 percent; at the temperature of 16 ℃, the tank pressure of 0.02MPa and the air flow of 1: 0.75 (v/v.min), with a stirring speed of 40rpm, for 4-5 days;
after the fermentation is started, taking a fermentation liquid sample every 22-24 hours, and measuring the pH value, reducing sugar and amino nitrogen: when the pH is less than 7, the reducing sugar is less than 1.5 percent, the amino nitrogen is less than 0.4mg/mL, and the bacterial concentration is not less than 14 percent, the fermentation is finished to obtain the paecilomyces hepiali fermentation culture solution;
the activity of beta-glucosidase in the paecilomyces hepiali fermentation culture solution is not less than 1200U/g.
Paecilomyces hepiali can produce beta-Glucosidase (beta-D-Glucosidase), which is also called beta-D-glucoside hydrolase, belongs to the class of cellulase, is an important component in a cellulolytic enzyme system, can be hydrolyzed and combined with a terminal non-reducing beta-D-glucose bond, and releases beta-D-glucose and corresponding aglycone. Beta-glucosidase is also a hydrolase with important physiological function, which can catalyze oxygen-affinitive glycosyl transfer, and can catalyze aryl, amino or alkyl-beta-D-glucoside and glycoside bonds in crude cyan glucoside, short chain oligosaccharide and disaccharide under certain conditions. According to this principle, SDG of linseed lignan can be converted into sec by this desugarization of β -glucosidase in industrial production process. Therefore, the absorption rate and the conversion rate of the linseed lignan in the body are greatly improved, and the greater biological activity is generated.
Preferably, in step S2, the ratio of the concentrated powder of flaxseed to water in parts by weight is 1: 2-4; the mass part ratio of the double-petal red rose pollen to the water is 1: 2-4; the mass part ratio of the gamma-aminobutyric acid to the water is 1: 2-4; the mass portion ratio of the red ginseng powder to the water is 1: 2-4; the mass part ratio of the red date powder to the water is 1: 2-4; the mass part ratio of the medlar powder to the water is 1: 2-4; the mass part ratio of the cranberry powder to water is 1: 2-4.
Preferably, in the step S3, the ratio of the volume of the paecilomyces hepiali fermentation culture solution to the total volume of the linseed dispersion solution, the red rose dispersion solution, the gamma-aminobutyric acid dispersion solution, the red ginseng dispersion solution, the red date dispersion solution, the medlar dispersion solution and the cranberry dispersion solution is 9: 1-3;
standing at 30-37 deg.C; the reaction time is 12-16 hours; 500-600r/min during centrifugation; the drying temperature is 85-130 ℃; the drying time is 10-15 min.
Preferably, in step S3, the final product is in powder form, and the particle size of the final product is 50-100 mesh.
The application of the fermented composition of Paecilomyces hepiali strain in the product for treating female endocrine dyscrasia is achieved by the fermented composition of Cordyceps militaris strain in any one of claims 1 to 3 or the fermented composition of Paecilomyces hepiali strain obtained by the preparation method in any one of claims 4 to 9.
The invention has the beneficial effects that:
the invention provides a paecilomyces hepiali strain fermentation composition, which can convert SDG in flaxseed lignans into SECO respectively by utilizing the desugarization of beta-glucosidase in a culture solution obtained after fermentation of the paecilomyces hepiali strain, wherein the SECO is easier to absorb than the SDG, so that the bioavailability of the flaxseed lignans is greatly improved and can be improved to 75% from 15%; under the action of beta-glucosidase, various glycosides or glycosides in other raw materials including red rose, gamma-aminobutyric acid, red ginseng, red date, medlar and cranberry raw materials can be converted into aglycone, the aglycone is easy to be absorbed by human bodies, and the bioavailability of active ingredients in the raw materials is further promoted to be remarkably improved.
The prescription mainly uses the traditional Chinese medicine to enrich and activate blood, tonify qi and nourish liver, improve metabolism capability, promote microcirculation in a body, accelerate internal circulation, and further play a role in conditioning female endocrine dyscrasia to improve the syndrome caused by female estrogen reduction.
Detailed Description
The present invention is further illustrated below with reference to specific examples. It will be appreciated by those skilled in the art that the following examples, which are set forth to illustrate the present invention, are intended to be part of the present invention, but not to be construed as limiting the scope of the present invention. The reagents used are all conventional products which are commercially available.
Example 1:
the strain source is as follows: paecilomyces hepiali (Paecilomyces hepiali) No.3.15540 was purchased from China general microbiological Collection management center, institute of microbiology, China academy of sciences. The preservation time is 2016, 2 and 19 days. The culture temperature is 25 ℃, and the collection state is as follows: china, collection site: yushu of Qinghai province.
A preparation method of a paecilomyces hepiali strain fermentation composition comprises the following steps:
s1, selecting paecilomyces hepiali powder according to a corresponding proportion, dispersing in water, and inoculating with an inoculation amount of 5 wt%. Inoculating the paecilomyces hepiali powder dispersion solution, firstly culturing by slant culture (slant culture medium: 10 wt% potato +1 wt% glucose +0.8 wt% peptone +0.1 wt% KH)2PO4+0.05wt％MgSQ4·7H2O +2 wt% agar, pH: 5.0-7.5; and (3) sterilizing a culture medium: 0.1MPa for 20 minutes; the paecilomyces hepiali powder is dispersed in water and then inoculated on a culture medium, and is cultured for 7 days at a constant temperature of 15 ℃, and then shake flask culture is carried out (shake flask culture medium: 1 wt% of grapeSugar, 0.2 wt% peptone, 0.4 wt% bran, 0.1 wt% KH2PO4、0.03wt％MgSQ4·7H2O, 0.03 wt% zinc sulfate, 0.1 wt% histidine, 0.05 wt% vitamin B1, pH: 5.0 to 7.5; under the aseptic condition, inoculating the strain cultured by the slant into a shake flask culture medium, and culturing for 7 days in a 15 ℃ constant temperature shaking table under the illumination condition; oscillating frequency of a reciprocating shaking table is 135 times/minute, red light with wavelength of 622-760 nm is used for irradiation in the whole shaking culture process, and then seeding tank culture is carried out (the composition of a culture medium is: 1 wt% hot-pressed soybean cake powder, 1 wt% glucose, 1 wt% sucrose, 0.1 wt% monopotassium phosphate, 0.05 wt% magnesium sulfate, 0.05 wt% zinc sulfate, 0.1 wt% histidine, 0.02 wt% vitamin B1, 0.1 wt% soybean oil, 0.05 wt% adenine, pH: 5.0 to 7.5; under the aseptic condition, inoculating the strain cultured in the shake flask into a seed tank by adopting a pressure difference method according to the inoculation amount of 3 percent, wherein the temperature is 16 ℃, the tank pressure is 0.02MPa, and the ventilation volume is 1: static culture for 4 days under the condition of 0.5 (v/v.min); culturing in a propagation tank (the culture medium comprises 1 wt% of hot pressed bean cake powder, 1 wt% of glucose, 1 wt% of sucrose, 0.1 wt% of monopotassium phosphate, 0.04 wt% of magnesium sulfate, 0.03 wt% of zinc sulfate, 0.1 wt% of histidine, 0.1 wt% of vitamin B1, 0.2 wt% of soybean oil, 0.05 wt% of adenine and 0.05 wt% of uracil, the pH value is 5.0-7.5), inoculating the cultured strain in the seed tank into the propagation tank by adopting a differential pressure method according to 9% of inoculation amount under the aseptic condition, and standing and culturing for 4 days under the conditions of 16 ℃ of temperature, 0.02MPa of tank pressure and 1: 0.15 (v/v.min) of ventilation amount; performing fermentation tank culture (culture medium comprises 1 wt% of hot pressed soybean cake powder, 1 wt% of glucose, 1 wt% of sucrose, 0.2 wt% of monopotassium phosphate, 0.04 wt% of magnesium sulfate, 0.04 wt% of zinc sulfate, 0.2 wt% of histidine, 0.2 wt% of vitamin B1, 0.2 wt% of soybean oil, 0.05 wt% of adenine and 0.05 wt% of uracil, wherein the pH is 5.0-7.5, inoculating strains cultured in a propagation tank into the fermentation tank by adopting a pressure difference method according to the inoculation amount of 5% under the aseptic condition, and culturing for 5 days under the conditions that the temperature is 16 ℃, the tank pressure is 0.02MPa, the ventilation volume is 1: 0.75 (v/v.min) and the stirring speed is 40 rpm);
after the start of the fermentation, samples of the fermentation broth were taken every 22 hours, and the pH, reducing sugar, amino nitrogen: when the pH is less than 7, the reducing sugar is less than 1.5 percent, the amino nitrogen is less than 0.4mg/mL, and the bacterial concentration is not less than 14 percent, the fermentation is finished, and the paecilomyces hepiali fermentation culture bacterial liquid is obtained.
The activity of beta-glucosidase in the paecilomyces hepiali fermentation culture solution is not less than 1200U/g.
(S2) selecting linseed concentrated powder (linseed concentrated powder wt%: water wt%: 1: 2), bivalvular red rose pollen (bivalvular red rose pollen wt%: water wt%: 1: 2), gamma-aminobutyric acid (gamma-aminobutyric acid wt%: 1: 2), red ginseng powder (red ginseng powder wt%: water wt%: 1: 2), red date powder (red date powder wt%: water wt%: 1: 2), medlar powder (medlar powder wt%: water wt%: 1: 2) and cranberry powder (cranberry powder wt%: water wt%: 1: 2) according to corresponding proportions, and adding water for dispersion to obtain linseed dispersion, bivalvular red rose dispersion, gamma-aminobutyric acid dispersion, red ginseng dispersion, red date dispersion, medlar dispersion and cranberry dispersion, respectively;
s3, mixing and uniformly mixing the volume of the paecilomyces hepiali fermentation culture solution with the total volume of 9:1 of flaxseed dispersion liquid, double red rose dispersion liquid, gamma-aminobutyric acid dispersion liquid, red ginseng dispersion liquid, red date dispersion liquid, medlar dispersion liquid and cranberry dispersion liquid, standing at 30 ℃ for reaction for 12 hours, centrifuging at 500r/min to obtain yellow semitransparent supernatant, and drying at 85 ℃ for 10min to obtain a finished product.
Example 2:
strain: paecilomyces hepiali (Cordyceps sinensis) No.5.825 is purchased from China general microbiological Collection management center, institute of microbiology, China academy of sciences. The preservation time is 1 month and 1 day in 2002. The culture temperature is 25 ℃, and the collection state is as follows: china, collection site: shaanxi.
A preparation method of a paecilomyces hepiali strain fermentation composition comprises the following steps:
s1, selecting paecilomyces hepiali powder according to a corresponding proportion, dispersing in water, and inoculating with an inoculation amount of 20 wt%. Inoculating the paecilomyces hepiali powder dispersion solution, firstly benefitingSlant culture (culture process: slant culture medium: 20 wt% potato +2 wt% glucose +1 wt% peptone +0.1 wt% KH)2PO4+0.05wt％MgSQ4·7H2O +4 wt% agar, pH: 5.0-7.5; and (3) sterilizing a culture medium: 0.2MPa for 30 minutes; the paecilomyces hepiali powder is dispersed in water and then inoculated on a culture medium, and is cultured for 9 days at a constant temperature of 20 ℃, and then shake flask culture is carried out (shake flask culture medium: 5 wt% glucose, 0.4 wt% peptone, 0.6 wt% bran, 0.3 wt% KH2PO4、0.05wt％MgSQ4·7H2O, 0.05 wt% zinc sulfate, 0.3 wt% histidine, 0.1 wt% vitamin B1, pH: 5.0 to 7.5; under aseptic condition, inoculating the strain cultured by the above slant into shake flask culture medium, and culturing in 20 deg.C constant temperature shaking table under illumination condition for 9 days; oscillating frequency of a reciprocating shaking table is 135 times/minute, red light with wavelength of 622-760 nm is used for irradiation in the whole shaking culture process, and then seeding tank culture is carried out (the composition of a culture medium is: 2 wt% hot-pressed soybean cake powder, 2 wt% glucose, 2 wt% sucrose, 0.2 wt% monopotassium phosphate, 0.05 wt% magnesium sulfate, 0.05 wt% zinc sulfate, 0.1 wt% histidine, 0.02 wt% vitamin B1, 0.1 wt% soybean oil, 0.05 wt% adenine, pH: 5.0 to 7.5; under the aseptic condition, inoculating the strain cultured in the shake flask into a seed tank by adopting a pressure difference method according to the inoculation amount of 3 percent, wherein the temperature is 16 ℃, the tank pressure is 0.02MPa, and the ventilation volume is 1: static culture for 5 days under the condition of 0.5 (v/v.min); culturing in a propagation tank (the culture medium comprises 2 wt% of hot pressed bean cake powder, 2 wt% of glucose, 2 wt% of sucrose, 0.1 wt% of monopotassium phosphate, 0.04 wt% of magnesium sulfate, 0.03 wt% of zinc sulfate, 0.1 wt% of histidine, 0.1 wt% of vitamin B1, 0.2 wt% of soybean oil, 0.05 wt% of adenine and 0.05 wt% of uracil, the pH value is 5.0-7.5), inoculating the strain cultured in the seed tank into the propagation tank by adopting a differential pressure method according to 9% of inoculation amount under the aseptic condition, and standing and culturing for 5 days under the conditions of 16 ℃ of temperature, 0.02MPa of tank pressure and 1: 0.15 (v/v.min) of ventilation amount; performing fermentation tank culture (culture medium: fermentation tank culture medium comprises 3 wt% of hot pressed bean cake powder, 1 wt% of glucose, 1 wt% of sucrose, 0.2 wt% of potassium dihydrogen phosphate, 0.04 wt% of magnesium sulfate, 0.04 wt% of zinc sulfate, 0.2 wt% of histidine, 0.2 wt% of vitaminsB1, 0.2 wt% soybean oil, 0.05 wt% adenine, 0.05 wt% uracil, pH: 5.0 to 7.5; under the aseptic condition, inoculating the cultured strains in the propagation tank into a fermentation tank by adopting a differential pressure method according to the inoculation amount of 5 percent; at the temperature of 16 ℃, the tank pressure of 0.02MPa and the air flow of 1: 0.75 (v/v.min), with a stirring speed of 40rpm, for 4 days;
after the start of the fermentation, samples of the fermentation broth were taken every 24 hours, and the pH, reducing sugar, amino nitrogen: when the pH is less than 7, the reducing sugar is less than 1.5 percent, the amino nitrogen is less than 0.4mg/mL, and the bacterial concentration is not less than 14 percent, the fermentation is finished, and the paecilomyces hepiali fermentation culture bacterial liquid is obtained.
The activity of beta-glucosidase in the paecilomyces hepiali fermentation culture solution is not less than 1200U/g.
(S2) selecting linseed concentrated powder (linseed concentrated powder wt%: water wt%: 1: 4), bivalvular red rose pollen (bivalvular red rose pollen wt%: water wt%: 1: 4), gamma-aminobutyric acid (gamma-aminobutyric acid wt%: 1: 4), red ginseng powder (red ginseng powder wt%: water wt%: 1: 4), red date powder (red date powder wt%: water wt%: 1: 4), medlar powder (medlar powder wt%: water wt%: 1: 4) and cranberry powder (cranberry powder wt%: water wt%: 1: 4) according to corresponding proportions, and adding water for dispersion to obtain linseed dispersion, bivalvular red rose dispersion, gamma-aminobutyric acid dispersion, red ginseng dispersion, red date dispersion, medlar dispersion and cranberry dispersion, respectively;
s3, the ratio of the volume of the paecilomyces hepiali fermentation culture solution to the total volume of the flaxseed dispersion liquid, the double-petal red rose dispersion liquid, the gamma-aminobutyric acid dispersion liquid, the red ginseng dispersion liquid, the red date dispersion liquid, the medlar dispersion liquid and the cranberry dispersion liquid is 9: 3, mixing, uniformly mixing, standing at 37 ℃ for 16 hours for reaction, centrifuging at 600r/min to obtain yellow semitransparent supernatant, and drying at 130 ℃ for 15 minutes to obtain a finished product.
Examples of the experiments
400 parts of linseed concentrated powder, 200 parts of rose powder with multiple petals, 200 parts of gamma-aminobutyric acid, 300 parts of red ginseng powder, 300 parts of red date powder, 300 parts of medlar powder, 300 parts of cranberry powder and 600 parts of paecilomyces hepiali powder are adopted as formula dosage. The finished solid particles obtained by the preparation method of example 2 were used as experimental reagents.
Method of volunteer experience efficacy:
(1) and (3) inclusion standard: 20-60 years old women have no obvious history of cardiovascular and cerebrovascular diseases and have no definite diabetic complications.
(2) The taking dosage is as follows: the medicine is taken after supper once every day, 1-2 bags (2 g/bag) are taken each time. The administration is continued for 21 days (during menstruation, the administration is avoided).
The experience result statistical method comprises the following steps:
remarking: symptom change and outcome assessment: and (4) judging a result: no change: no progression, such as from 2 to 2; the improvement is as follows: up to one stage, for example from 1 to 2: obviously improve the following steps: and two stages, for example from 1 to 3. Wherein, Q: before the medicine is taken; h: this is indicated after administration.
Change of symptoms:
menstruation volume: no I, rare II, normal III and high IV.
Menses properties: 1. dark brown, 2. dark red, 3. bright red, 4. with blood clots.
Monthly menstrual days: 1: 0,2: < 2 days, 3: day > 3, 4: and more than 5 days.
Dysmenorrhea conditions: 1: none, 2: mild, 3: medium, 4: and (4) heavy.
Emotional state: 1. frequent emotional restlessness, 2. easy agitation, and 3. stable mood.
Climacteric symptoms: flushing, headache, dizziness, fatigue, weakness, sweating, etc. on the face: 1. frequently, 2. less frequently, 3. none.
A sleep state: 1. not easy to fall asleep, 2, easy to wake up and 3, good sleep.
Skin condition: 1. loose and poor elasticity, 2. plump and good elasticity.
Vaginal lubrication status during sexual life: 1. dry and astringent, 2. lubricate.
Breast state: 1. lobular hyperplasia, 2. with nodules of the breast, 3. mastoptosis, 4. with elasticity.
Uterine status: 1. hysteromyoma, 2, endometriosis, 3, no abnormalities.
As a result: the product has an improvement rate (improvement + obvious improvement) of 88% for users, and has the effect of remarkably improving female endocrine dyscrasia so as to improve the syndrome caused by female estrogen reduction.
The present invention is not limited to the above alternative embodiments, and any other products in various forms can be obtained by the present invention, and the present invention is within the protection scope of the present invention. The above embodiments should not be construed as limiting the scope of the present invention, and it will be understood by those skilled in the art that modifications may be made to the technical solutions described in the above embodiments, or equivalent substitutions may be made to some or all of the technical features thereof, without departing from the scope of the present invention, and at the same time, such modifications or substitutions may not make the essence of the corresponding technical solutions depart from the scope of the embodiments of the present invention.
Claims (10)
1. A paecilomyces hepiali strain fermentation composition is characterized by being prepared from the following raw materials in parts by mass:
20-60 parts of linseed concentrated powder, 10-30 parts of rose powder with double petals, 10-30 parts of gamma-aminobutyric acid, 10-55 parts of red ginseng powder, 10-55 parts of red date powder, 10-55 parts of medlar powder, 10-55 parts of cranberry powder and 20-100 parts of paecilomyces hepiali powder.
2. The paecilomyces hepiali strain fermentation composition according to claim 1, wherein the composition is prepared from the following raw materials in parts by mass:
30-50 parts of linseed concentrated powder, 15-25 parts of rose powder with double petals, 15-25 parts of gamma-aminobutyric acid, 22-45 parts of red ginseng powder, 22-45 parts of red date powder, 22-45 parts of medlar powder, 22-45 parts of cranberry powder and 44-80 parts of paecilomyces hepiali powder.
3. The paecilomyces hepiali strain fermentation composition according to claim 2, wherein the composition is prepared from the following raw materials in parts by mass:
40 parts of linseed concentrated powder, 20 parts of rose powder with double petals, 20 parts of gamma-aminobutyric acid, 30 parts of red ginseng powder, 30 parts of red date powder, 30 parts of medlar powder, 30 parts of cranberry powder and 60 parts of paecilomyces hepiali powder.
4. A process for the preparation of a paecilomyces hepiali strain fermentation composition as defined in any one of claims 1 to 3, said process comprising the steps of:
s1, selecting paecilomyces hepiali powder according to a corresponding proportion, adding water for dispersion, performing strain culture, and performing strain fermentation culture to obtain a paecilomyces hepiali fermentation culture solution;
s2, selecting linseed concentrated powder, double-petal red rose pollen, gamma-aminobutyric acid, red ginseng powder, red date powder, medlar powder and cranberry powder according to a corresponding proportion, and adding water for dispersing to respectively obtain linseed dispersion liquid, double-petal red rose dispersion liquid, gamma-aminobutyric acid dispersion liquid, red ginseng dispersion liquid, red jujube dispersion liquid, medlar dispersion liquid and cranberry dispersion liquid;
s3, mixing the paecilomyces hepiali fermentation culture solution, the linseed dispersion solution, the double red rose dispersion solution, the gamma-aminobutyric acid dispersion solution, the red ginseng dispersion solution, the red date dispersion solution, the medlar dispersion solution and the cranberry dispersion solution, uniformly mixing, standing for reaction, centrifuging and drying to obtain a finished product.
5. The method for preparing fermented composition of Paecilomyces hepiali strain according to claim 4, wherein in step S1, when the Paecilomyces hepiali powder is inoculated after being dispersed in water, the inoculation amount is 2.5-5 wt% of the Paecilomyces hepiali powder in water dispersion liquid;
the strain culture comprises strain slant culture and strain shake flask culture;
the strain slant culture process comprises the following steps: the composition of slant culture medium includes 2-4 wt% of glucose, 0.2-0.4 wt% of peptone, 0.4-0.8 wt% of bran, 0.1-0.3 wt% of potassium dihydrogen phosphate, 0.03-0.05 wt% of magnesium sulfate, 0.1-0.3 wt% of histidine, 2-4 wt% of agar, pH: 5.0 to 7.5; and (3) sterilizing a culture medium: 0.1MPa for 30-40 min; inoculating the strain to a slant culture medium, and culturing at 15-20 deg.C for 7-9 days;
the strain shake flask culture process is as follows: the shake flask culture medium comprises 1-5 wt% of glucose, 0.2-0.4 wt% of peptone, 0.4-0.6 wt% of bran, 0.1-0.3 wt% of monopotassium phosphate, 0.03-0.05 wt% of magnesium sulfate, 0.03-0.05 wt% of zinc sulfate, 0.1-0.3 wt% of histidine, 10.05-0.1 wt% of vitamin B, and pH: 5.0 to 7.5; inoculating cultured strain into shake flask culture medium under aseptic condition, and culturing in 15-20 deg.C constant temperature shaking table under illumination for 7-9 days; the oscillating frequency of the reciprocating shaking table is 135 times/minute, and red light with the wavelength of 622-760 nm is used for irradiation in the whole shaking culture process.
6. The method for preparing a fermented composition of Paecilomyces hepiali strain according to claim 4, wherein in step S1, the strain fermentation culture comprises seed tank culture, propagation tank culture and fermenter culture;
the culture medium of the seeding tank comprises the following components: 1-2 wt% of hot-pressed soybean cake powder, 1-2 wt% of glucose, 1-2 wt% of sucrose, 0.1-0.2 wt% of monopotassium phosphate, 0.05 wt% of magnesium sulfate, 0.05 wt% of zinc sulfate, 0.1 wt% of histidine, 0.02 wt% of vitamin B1, 0.1 wt% of soybean oil, 0.05 wt% of adenine, pH: 5.0 to 7.5; under the aseptic condition, inoculating the strain cultured in the shake flask into a seed tank by adopting a pressure difference method according to the inoculation amount of 3 percent, wherein the temperature is 16 ℃, the tank pressure is 0.02MPa, and the ventilation volume is 1: standing and culturing for 4-5 days under the condition of 0.5 (v/v.min);
the propagation tank culture medium comprises the following components: 1-2 wt% of hot-pressed soybean cake powder, 1-2 wt% of glucose, 1-2 wt% of sucrose, 0.1 wt% of monopotassium phosphate, 0.04 wt% of magnesium sulfate, 0.03 wt% of zinc sulfate, 0.1 wt% of histidine, 0.1 wt% of vitamin B1, 0.2 wt% of soybean oil, 0.05 wt% of adenine, 0.05 wt% of uracil, pH: 5.0 to 7.5; under the aseptic condition, inoculating the cultured strains in the seed tank into a propagation tank by adopting a differential pressure method according to 9 percent of inoculation amount; at the temperature of 16 ℃, the tank pressure of 0.02MPa and the air flow of 1: standing and culturing for 4-5 days under the condition of 0.15 (v/v.min);
the culture medium of the fermentation tank: the composition of the fermenter medium was: 1-3 wt% hot-pressed soybean cake powder, 1 wt% glucose, 1 wt% sucrose, 0.2 wt% monopotassium phosphate, 0.04 wt% magnesium sulfate, 0.04 wt% zinc sulfate, 0.2 wt% histidine, 0.2 wt% vitamin B1, 0.2 wt% soybean oil, 0.05 wt% adenine, 0.05 wt% uracil, pH: 5.0 to 7.5; under the aseptic condition, inoculating the cultured strains in the propagation tank into a fermentation tank by adopting a differential pressure method according to the inoculation amount of 5 percent; at the temperature of 16 ℃, the tank pressure of 0.02MPa and the air flow of 1: 0.75 (v/v.min), with a stirring speed of 40rpm, for 4-5 days;
after the fermentation is started, taking a fermentation liquid sample every 22-24 hours, and measuring the pH value, reducing sugar and amino nitrogen: when the pH is less than 7, the reducing sugar is less than 1.5 percent, the amino nitrogen is less than 0.4mg/mL, and the bacterial concentration is not less than 14 percent, the fermentation is finished to obtain the paecilomyces hepiali fermentation culture solution;
the activity of beta-glucosidase in the paecilomyces hepiali fermentation culture solution is not less than 1200U/g.
7. The method for preparing a fermented composition of Paecilomyces hepiali strain according to claim 5 or 6, wherein in step S2, the ratio of the concentrated powder of flax seeds to the water in parts by mass is 1: 2-4; the mass part ratio of the double-petal red rose pollen to the water is 1: 2-4; the mass part ratio of the gamma-aminobutyric acid to the water is 1: 2-4; the mass portion ratio of the red ginseng powder to the water is 1: 2-4; the mass part ratio of the red date powder to the water is 1: 2-4; the mass part ratio of the medlar powder to the water is 1: 2-4; the mass part ratio of the cranberry powder to water is 1: 2-4.
8. The method for preparing a fermented composition of Paecilomyces hepiali strain according to claim 4, wherein in step S3, the ratio of the volume of the fermented culture solution of Paecilomyces hepiali to the total volume of the linseed dispersion, the red rose dispersion, the gamma-aminobutyric acid dispersion, the red ginseng dispersion, the red date dispersion, the medlar dispersion and the cranberry dispersion is 9: 1-3;
standing at 30-37 deg.C; the reaction time is 12-16 hours; 500-600r/min during centrifugation; the drying temperature is 85-130 ℃; the drying time is 10-15 min.
9. The method for preparing a fermented composition of Paecilomyces hepiali strain according to claim 4, wherein in step S3, the final product is in powder form, and the particle size of the final product is 50-100 mesh.
10. The application of the paecilomyces hepiali strain fermentation composition is characterized in that the cordyceps militaris strain fermentation composition as defined in any one of claims 1 to 3 or the paecilomyces hepiali strain fermentation composition obtained by the preparation method as defined in any one of claims 4 to 9 is applied to female endocrine dyscrasia products.
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| 王彩霞，等: "玫参胶囊对去卵巢大鼠生殖内分泌系统影响的实验研究", 《新中医》 * |
| 雏菊: "红枣枸杞玫瑰花茶的功效与作用", 《百度网页频道》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP7286861B1 (en) | 2022-11-16 | 2023-06-05 | 三和酒類株式会社 | Health maintenance agent for mammary gland tissue containing GABA as an active ingredient |
| CN116211938A (en) * | 2023-03-06 | 2023-06-06 | 无锡康能特医食品科技有限公司 | Lentinus edodes strain fermentation composition and preparation method and application thereof |
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