CN102428832A - Fungal pharmaceutical mycoplasm with blood sugar lowering efficacy and preparation method thereof - Google Patents
Fungal pharmaceutical mycoplasm with blood sugar lowering efficacy and preparation method thereof Download PDFInfo
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Abstract
The invention belongs to the field of biofermentation engineering. A pharmaceutical medium and a fungal strain which have blood sugar lowering efficacy are used as bidirectional fermentation raw materials, wherein the pharmaceutical medium is mainly composed of the following raw materials: cornel, gynostemma, momordica grosvenori, winged euony twigs, balsam pear and polyrhachis vicina; the fungal strain is composed of the following strains: inonotus obliquus, cordyceps, phellinus igniarius, polystictus versicolor, and grifola frondosa; and bi-directional multi-fungi fermentation is carried out between multiple edible and pharmaceutical fungi and Chinese herbal medicines by a bidirectional fermentation process, thus organisms in two kingdoms or three kingdoms are organically combined to obtain an entirely new blood-sugar-lowering pharmaceutical mycoplasm product. The obtained product can generate 1+1>2 physiological function efficiency for hypoimmunity, has the actions of obviously lowering blood sugar but not increasing insulin concentration, has significant positive effects on helping treating and controlling diabetic complications, and can be used as an oral medicament for preventing and treating diabetes or a health-care product for adjusting blood sugar.
Description
Technical field
The invention belongs to the biological fermentation engineering field, be specifically related to a kind of fungi fermentation medicinal fungal substance with effect of lowering blood sugar.
Background technology
The treating diabetes difficulty is big, and expense is high, has become the global problem that national governments pay close attention to.The medicine of specially for treating diabetes has only six or seven kinds on China's medical market at present, is divided into three major types: biological medicine insulin, chemical medicine sulfonylurea biguanides, and Chinese patent drug class.Insulin preparation costs an arm and a leg, and is difficult to promote and have 8 big side effects; Though chemistry medicine blood sugar reducing function is obvious, and is bigger to toxic and side effects such as liver, kidneys, also causes that cardiovascular complication increases the weight of easily etc.Though Chinese patent drug has certain therapeutic action, present clinical use be mostly crude extract preparation or doctor from the prescription decoction of medicinal ingredients of intending, lack drug effect significantly, active ingredient and the mechanism of action is clear, quality controllable, preparation is advanced modern Chinese herbal medicine.At present, the research range of Chinese patent drug is many in following ranges, has the problems referred to above equally:
One type is with different herbal prescriptions, or the Chinese patent drug that oven dry is pulverized or decocting is produced by conventional method.
Like patent CN02157912.1 " ant glucose in urine ball and processing method thereof ", with genseng, ant, cicada moves back, and traditional Chinese medicines such as the coptis are batching, and after oven dry pulverizing, decocting boiling down contracted, mixed powder and soup were processed particle.
Patent 200910094733 " a kind of Chinese medicinal capsule of treating diabetes " is characterized in that containing 23 kinds of Chinese herbal medicines such as green leaf chrysanthemum, imposing, gall nut, sealwort, American Ginseng.
Patent (AP) 02133230 " a kind of Yusanxiao medicine of treating diabetes and preparation method＞" be with Radix Angelicae Sinensis, the coptis, red ginseng, pilose antler respectively pulverizing mix with the water extract of 13 flavor Chinese medicines such as the Radix Astragali, glutinous rehmannia, prepared rhizome of rehmannia.
Patent 201010188883 " a kind of medicine of treating diabetes and preparation method thereof " is a bulk drug by following medicine: radix bupleuri, the root of herbaceous peony, the rhizome of nutgrass flatsedge, moutan bark etc., use Chinese medicine with coarse food grain mixture and nutrients compatibility.
Second type is to be main with Chinese medicinal plant, and traditional method processing is equipped with the synthetic drug of chemical medicine composition.
Belong to medicinal preparation like patent CN97103536.9 " a kind of medicine of treating diabetes and preparation method thereof ", inventive features is by kinds of traditional Chinese medicines: pig a kind of reed mentioned in ancient books, rhizoma alismatis, golden cypress, glutinous rice bud point etc. dissolve Metformin hydrochloride through drying frying, sugar is fitted flat.
The 3rd type is the compound medicine with the Chinese medical extract compatibility.
Mainly form like patent CN02138362.6 " a kind of Chinese medicine hypoglycemic agent and preparation method thereof " by the ethyl acetate extract or the extract of Ramulus euonymi.This drug main of patent 200610066305 " a kind of pharmaceutical composition of treating diabetes and preparation method thereof " will add chromium picolinate through the alcohol reflux extraction by tartarian buckwheat, mulberry leaf combination and process.
Up to now, the research that the edible and medicinal fungi biologic product is used to treat diabetes and complication aspect thereof is still few, the medicinal fungal substance goods that more useless two-way fermentation engineering obtains." the medicinal fungi biologic product and the preparation technologies thereof of treatment diabetes " are formulated by the extract of wild mushroom extract, mulberry leaf and Chinese yam mixture like patent 200810064886.Patent ZL03111130.0 " edible and medicinal fungi solid fermentation bacterium powder " also only limits to do the mycelium that solid fermentation obtains with nutrient matrix and forms through compatibility.
Summary of the invention
The present invention adopts is the two-way fermentation technique of microorganism and " many bacterium " fermentation technique of not relating to as yet of diabetes field so far.Promptly do property of medicine matrix, between many edible and medicinal fungis and Chinese herbal medicine, carry out amphicheirality's mixed culture fermentation, make the brand-new hypoglycemic property of medicine mycoplasma that organically combines between two or three boundary's biologies and obtain with Chinese herbal medicine.
Theoretical foundation of the present invention is following:
A: two-way fermentation engineering is meant; Fermentation substrate all adopts Chinese herbal medicine (or adding an amount of agricultural byproducts therein) and constitutes property of medicine matrix; Use medicinal fungi fermentation again, cause fermentation to have the amphicheirality, its tunning transfers medicinal fungal substance to by simple fungal mycelium.This medicinal fungal substance ratio just sends out (original) fungi or property of medicine matrix itself or both are added with better drug effect simply mutually, can produce the physiological function effectiveness of 1+1＞2, can significantly strengthen its active component, improves effects such as drug effect and curative effect.
The two-way zymotechnique that the present invention adopts; Be that fermented bacterium and property of medicine matrix are constituted in the fermentation combination; Its property of medicine matrix is changed into and is adopted the traditional Chinese medicine with certain hypoglycemic activity composition as property of medicine matrix by only used the agricultural byproducts that are rich in nutrition such as carbon, nitrogen as nutrient matrix in the past, and it can provide the growth desired nutritional for the fungi with hypoglycemic effect; Enzyme because of fungi is changed structural constituent again, thereby produces new nature,taste and action.Because of the present invention carries out the conversion of organic active composition and combines between two boundaries or three boundary's biologies, its functional component is abundanter again, and hypoglycemic effect is more obvious.
Technological process of the present invention is following:
Take by weighing property of medicine base starting material → add water → sterilization → pro-rata property of medicine matrix → insert respectively fungi strain → all medicinal fungal substances of two-way fermentation → collection → oven dry pulverizing → acquisition medicinal fungal substance goods.
B: adopt multiple edible and medicinal fungi with remarkable hypoglycemic activity composition and no side effects as fermented bacterium: Inonotus obliquus (Inonqqus obliquus); Cordyceps sinensis [Cordyceps sinensis (Berk.) Sacc]; Mulberry yellow [(pHellinus igniariusL.es Fr.) Quel.], grifola frondosus (Grifola frondosa), rainbow conk [Coriolus versicolor (L.:Fr.)].Above-mentioned bacterial classification all derives from Dalian, Liaoning Province precious gill fungus fungus research and development centre.
The function of blood sugar reduction of the various fungies of adopting is following:
Inonotus obliquus: Inonotus obliquus is at Russia's medicinal fungi as the treatment disease among the people, and long-term animal experiment and clinical testing show without any side effects, and hepatitis, hypertension are also had control efficiency.Ceng Xiaolong's, " chemical composition of Inonotus obliquus and pharmacological research " shows, its artificial culture mycelium contains a large amount of hypoglycemic, vegetable fiber class polysaccharide bodies of bringing back to life immunization, and the cure rate of diabetes is reached 93%.
Cordyceps sinensis: according to " the hypoglycemic mechanism researchs of edible fungi polysaccharide " such as Du Mei; Its main medicinal ingredient cordycepin has inhibitory action to hyperglycaemia; Two kinds of polysaccharide Chinese caterpillar fungus hypha heteroglycan CsOH (being the heteroglycan that contains L-arabinose, L-wood sugar, D-mannose, D-galactose and D-glucose) and polysaccharide semifinished product CHWc can significantly improve (containing D-mannose, D-galactose and D-glucose) physiologically active of beta Cell of islet, and hyperglycaemia is had tangible reduction effect.And can protect kidney to improve the immunity of human body, effectively control the various complication of diabetes.
Mulberry is yellow: the IL-1 cytokine that the yellow polysaccharide of mulberry is excited has the promotion insulin secretion, the effect of blood sugar lowering.Contain rich in amino acid and zinc simultaneously, have the effect of activation pancreas function.And can effectively improve the immunocompetence of patient of diabetes.The yellow bacterium of mulberry is promoted cell function, cholesterol reducing in addition, antithrombotic, and the effects such as contraction of mitigation blood vessel all help diabetic's blood pressure normalization (seeing " the domestic and international progress that rare medicinal fungi mulberry is yellow " such as Sun Pei dragons).
Disclose among grifola frondosus: the Du Mei etc. " the hypoglycemic mechanism research of edible fungi polysaccharide ", the functional component of grifola frondosus mainly is polysaccharide and glycoprotein, and β-(1, the 3)-D-glucan for a kind of originated from fungus has the function of blood sugar lowering.Higher Organic Chromium and the organic selenium of content has the effect of assisting insulin to keep normal sugar tolerance simultaneously in the grifola frondosus.Also have the active cell immunologic function in addition, bring high blood pressure down, the function of blood fat and serum insulin.Cirrhosis, diabetes are all produced effect and without any side effects.
Rainbow conk: (polysaccharide of Corlolus versicolor PSK) has control blood sugar rising effect to contained polysaccharide in the rainbow conk, and euglycemia is not had influence.Oral medication does not have the long term toxicity reaction.Experiment shows that PSK is the very low material of a toxicity, does not have mutagenesis (seeing " the hypoglycemic mechanism researchs of edible fungi polysaccharide " such as Du Mei) yet.
C: choose have that effect of reducing blood sugar is wide with, the crowd that is suitable for, the Chinese herbal medicine or the animal that have no side effect, normal blood glucose value is not had influence (not reducing) be as property of medicine matrix:
Dogwood: the contained total terpene of dogwood can improve the normal mouse sugar tolerance, obviously reduces the blood glucose value that alloxan is induced, and improves serum insulin level; Significantly reduce the blood glucose value of Streptozotocin diabetes model, increase hepatic glycogen content.Normal blood sugar level there is not obvious influence.Classified as one of medicinal and edible Chinese medicine material kind (seeing " Chinese medicine blood-sugar decreasing active research recent developments " such as Liang Lei) by country.
Gynostemma pentaphylla: according to Xu Shiming; " research of gynostemma pentaphylla and application " record of Guo Yuxiao; The hypoglycemic activity of Gynostemma pentaphylla polysaccharide is different with general antidiabetic drug; It can delay glucose in the absorption of enteron aisle, suppress alpha-glucosidase activity, can reduce the hyperglycaemia sufferer that alloxan induces and improve glycometabolism.Euglycemia is not had influence, protect suprarenal gland, thymus gland and endocrine organ's effect in addition, hypotensive blood fat, control complication.
Momordica grosvenori: Qi Xiangyang etc. " Momordica grosvenori is to the hypoglycemic activity of diabetic mice " research shows that it contains abundant glucoside, and the T2DM-II type diabetes that this glucoside is induced alloxan have remarkable hypoglycemic activity, and normal mouse blood sugar is not had influence.Prevent and treat hypertension, reducing blood lipid and antiobesity action in addition, the controlled complication.
Report in " the Ramulus euonymi pharmaceutical research is brief talked " of Ramulus euonymi: Yin Guiyu, Yang Jinping, it contains the composition of multiple blood sugar lowering, can obviously reduce the blood glucose value that alloxan is induced, and normal mice blood sugar is not had influence.Its oxaloacetic acid sodium composition can stimulate the beta cell hyperplasia, adjusts abnormal metabolic process, strengthens secretion of insulin.Nontoxic, take for a long time or heavy dose of (below the 30g) being decocted in water for oral dose does not have apparent side effect yet.Can also treat diseases such as coronary heart disease, hypertension, arteriosclerosis, the complication of diabetes is had control action.
Balsam pear: its momordica saponins is the main active ingredient of hypoglycemic, can obviously reduce alloxan diabetes patient's blood sugar, and total effective rate reaches 82% and do not have a toxicity (seeing beam thunder, limit Po Lam etc. " Chinese medicine blood-sugar decreasing active research recent developments ").
Polyhachis vicina Roger: there are some researches show that it is to have the new resource food that medicine-food two-purpose is worth, and is the bidirectional immune regulator that can keep the organism physiology balance, can two-way adjustment blood sugar and blood pressure.Contained biologically active chromium can assist insulin to keep sugar tolerance." cursive script formaldehyde " and contained extraordinary insect protein; Can stimulate body to produce protection antibody forcefully, can make the diabetic avoid the infringement (seeing " researchs of several kinds of animal Chinese medicine hypoglycemic activities " such as the Ren Geng that relaxes) of various organs with sterol and triterpene compound.
The present invention specifically realizes in the following manner:
One side of the present invention is: the present invention relates to a kind of raw material, comprises property of medicine matrix and fungi strain with fungi medicinal fungal substance of effect of lowering blood sugar, wherein:
Property of medicine matrix is made up of following raw material: dogwood, Gynostemma pentaphylla, Momordica grosvenori, Ramulus euonymi, balsam pear, Polyhachis vicina Roger, nutrient matrix; Wherein, the source of nutrient matrix is cereal, beans, rice bran, wheat bran;
Fungi strain is made up of following bacterial classification: Inonotus obliquus, Cordyceps sinensis, the yellow bacterium of mulberry, rainbow conk, grifola frondosus;
The inoculation weight of fungi strain is 8～12wt% of property of medicine matrix.
In the raw material of above-mentioned fungi medicinal fungal substance, when property of medicine matrix and fungi strain during in following ratio proportioning, effect is preferable:
Property of medicine matrix is mixed by the raw material of following drying by mass percentage:
The corresponding relation of each fungi strain and used property of medicine matrix is respectively by mass percentage:
The inoculation weight of fungi strain is 8～12wt% of property of medicine matrix.
In the raw material of above-mentioned fungi medicinal fungal substance, when property of medicine matrix and fungi strain during in following ratio proportioning, best results:
Property of medicine matrix is mixed by the raw material of following drying by mass percentage:
The corresponding relation of each fungi strain and used property of medicine matrix is respectively by mass percentage:
The inoculation weight of fungi strain is the 10wt% of property of medicine matrix.
Other one side of the present invention is: a kind of preparation method with fungi medicinal fungal substance of effect of lowering blood sugar, and its concrete processing step is following:
(e) take by weighing dry property of medicine base starting material and mixing by mass percentage;
(f) in the mixture of step a gained, add the running water of pH7～8 of 1～10 times of mixture quality, in 100～127 ℃ of sterilization 1～10h, obtain property of medicine matrix;
(g) by the corresponding relation of each fungi strain and used property of medicine matrix; The property of medicine matrix of allocation step b gained inserts fungi strain respectively under aseptic condition, inoculative proportion is 8～12wt% of property of medicine matrix; 23～27 ℃ of two-way fermentations 4～45 days, obtain medicinal fungal substance respectively;
(h) collect all medicinal fungal substances that step c obtains, mix obtaining this product after oven dry is pulverized.
Concrete: among the preparation method of above-mentioned fungi medicinal fungal substance, the two-way fermentation mode of employing is solid fermentation or liquid deep layer fermenting.
Concrete: among the preparation method of above-mentioned fungi medicinal fungal substance, the liquid deep layer fermenting condition is: 25～27 ℃ of temperature, tank pressure 0.7～0.8kg/cm
2, throughput 1: 1v/v/ branch.
Concrete: among the preparation method of above-mentioned fungi medicinal fungal substance, the inoculative proportion of said fungi strain is the 10wt% of property of medicine matrix.
Other one side of the present invention is: through the fungi medicinal fungal substance of method for preparing.
Other one side of the present invention is: the application of above-mentioned fungi medicinal fungal substance in preparation electuary, pill, tablet, capsule.
The liquid deep layer fermenting technology of edible mushroom belongs to one of modern biotechnology.Submerged fermentation technology Direct Production edible mushroom thalline, the zymotic fluid of nutrient components such as amino acid and secondary metabolites is rich in acquisition simultaneously.The key of edible fungus liquid submerged fermentation technical research is that different edible mushrooms will be cultivated with different fermentation substrates, therefore, select and prepare the key that fermentation substrate is an edible fungus liquid submerged fermentation technology.Utilize food (medicine) can obtain a large amount of mycelium and tunning thereof within a short period of time,, therefore in Edible Fungi, have broad application prospects because this process cycle is short, output is high, cost is low, process equipment is simple with the bacteria liquid submerged fermentation.Food (medicine) can produce the several physiological active substances such as multiple compound and plant hormone of polysaccharide, alkaloid, terpenoid, sterol, enzyme, nucleic acid, vitamin, the effect of tool antibiotic in the submerged fermentation process with fungi, these materials have respectively effects such as the effect of preventing and curing diseases of human organs such as cardiovascular, liver, nervous system, kidney, property and anticancer, anti-inflammatory, anti-ageing, antibiotic, raising immunity.
The product that the present invention relates to adopts edibleization that organise; Pollution-free property of medicine matrix that has no side effect and various fungi; And in whole production technology, do not add and use any chemical substance,, avoid in treatment and health care, human body being formed the secondary injury to guarantee security of products.
The present invention organically combines the composition of the interbiotic main hypoglycemic factor cordycepin in three boundaries, multiple fungi polysaccharide and multiple medicinal plant polysaccharide and biological chromium through amphicheirality's fermentation and physiological conversion, forms to have the new bioactive molecule group and the medicinal fungal substance of secondary metabolites.
Beneficial effect: the invention belongs to biomedicine field.Two-way fermentation technique of microorganism that the diabetes field so far of adopting does not relate to as yet and mixed culture fermentation technology make to organically combine between two or three boundary's biologies and obtain brand-new hypoglycemic mycoplasma.This product has the effect that tangible blood sugar lowering does not increase insulin concentration again, for the complication of supplemental treatment and control diabetes obvious and positive effect is arranged, and the acute hypoglycemia that does not cause common medicine and produced.Quality controllable than existing blood-sugar lowering tcm drug, formulation is advanced.This product belongs to pure biologic product, and is without any side effects; Said preparation is suitable for diabetic's treatment, the prevention of diabetes, can be used as the medicinal preparation for oral administration of blood sugar lowering or the health medicine of adjusting blood sugar.
Embodiment
The raw material sources that relate in the following non-limiting example are following:
Dogwood, Gynostemma pentaphylla, Momordica grosvenori, Ramulus euonymi, all purchase in the shop of Chinese medicines;
Balsam pear is purchased the Yi Lin Food Co., Ltd in Shanghai;
Polyhachis vicina Roger derives from Heng County, Guangxi pure ant powder factory;
Fermented bacterium: Inonotus obliquus (Inonqqus obliquus); Cordyceps sinensis [Cordyceps sinensis (Berk.) Sacc]; Mulberry yellow [(pHellinus igniariusL.ex Fr.) Quel.], grifola frondosus (Grifola frondosa), rainbow conk [Coriolus versicolor (L.:Fr.)].Above-mentioned bacterial classification all derives from Dalian, Liaoning Province precious gill fungus fungus research and development centre;
Laboratory animal: kunming mice, small white mouse, body weight 18～22g, male and female half and half are purchased the good universe journey bio tech ltd in Shenyang.
Following non-limiting example can make those of ordinary skill in the art more fully understand the present invention, but does not limit the present invention in any way.
Embodiment 1
(1) preparation of medicinal fungal substance
(a) with Chinese herbal medicine after 50～60 ℃ of oven dry, take by weighing dry property of medicine base starting material and mix by following mass percent: dogwood 2000g, Gynostemma pentaphylla 3000g, Momordica grosvenori 1500g, Ramulus euonymi 1500g, balsam pear 1500g, Polyhachis vicina Roger 500g.Above-mentioned raw materials is crushed to 1～2mm.
(b) with the mixture of the step a gained container of packing into, add the running water of pH7.5 by 1: 1 weight ratio, in 127 ℃ of sterilization 2h, be cooled to below 30 ℃ after, obtain property of medicine matrix.
(c) under aseptic condition, in 2000g property of medicine matrix, insert the Inonotus obliquus bacterial strain; In 4000g property of medicine matrix, insert Cordyceps strain; In 2000g property of medicine matrix, insert the yellow bacteria strain of mulberry; In 1000g property of medicine matrix, insert Grifola frondosa strain; In 1000g property of medicine matrix, insert the rainbow conk bacterial strain; The inoculative proportion of above-mentioned bacterial classification is 10% of the property of medicine matrix weight inoculated.
(d) connect rearmounted 25 ± 1 ℃ of fermentations of bacterium 45 days.
(e) collect all medicinal fungal substances that steps d obtains, dry to water content below 5% through 40～50 ℃, mix, obtain the medicinal fungal substance goods after nano cell broken wall cracker is crushed to below the 100nm.
(2) finished product detects
According to appendix VID of Pharmacopoeia of People's Republic of China version in 2005, high performance liquid chromatography is measured cordycepin and is reached 3.88mg/g; Measuring total polysaccharides according to UV-VIS spectrophotometry (appendix VA of Chinese Pharmacopoeia version in 2005) is 16.7%; According to chromium Determination on content method in standard GB/T5009.123-2003 food, measuring biological chromium content is 0.86mg/kg.
Embodiment 2
(1) preparation of medicinal fungal substance
(a) with Chinese herbal medicine after 50～60 ℃ of oven dry, take by weighing dry property of medicine base starting material and mix by following mass percent: dogwood 1000g, Gynostemma pentaphylla 2500g, Momordica grosvenori 1000g, Ramulus euonymi 2000g, balsam pear 3000g, Polyhachis vicina Roger 500g.Above-mentioned raw materials is crushed to 1～2mm.
(b) with the mixture of the step a gained container of packing into, add the running water of pH7.5 by 1: 1 weight ratio, in 127 ℃ of sterilization 2h, be cooled to below 30 ℃ after, obtain property of medicine matrix.
(c) under aseptic condition, in 3000g property of medicine matrix, insert the Inonotus obliquus bacterial strain; In 3000g property of medicine matrix, insert Cordyceps strain; In 2000g property of medicine matrix, insert the yellow bacteria strain of mulberry; In 1000g property of medicine matrix, insert Grifola frondosa strain; In 1000g property of medicine matrix, insert the rainbow conk bacterial strain; The inoculative proportion of above-mentioned bacterial classification is 10% of the property of medicine matrix weight inoculated.
(d) connect rearmounted 25 ± 1 ℃ of fermentations of bacterium 45 days.
Collect all medicinal fungal substances that steps d obtains, dry to water content below 5% through 40～50 ℃, mix, obtain the medicinal fungal substance goods after nano cell broken wall cracker is crushed to below the 100nm.
(2) finished product detects
According to appendix VID of Pharmacopoeia of People's Republic of China version in 2005, high performance liquid chromatography is measured cordycepin and is reached 3.3mg/g; Measuring total polysaccharides according to UV-VIS spectrophotometry (appendix VA of Chinese Pharmacopoeia version in 2005) is 14.6%; According to chromium Determination on content method in standard GB/T5009.123-2003 food, measuring biological chromium content is 0.89mg/kg.
Embodiment 3
(1) preparation of medicinal fungal substance
(a) with Chinese herbal medicine after 50～60 ℃ of oven dry, take by weighing dry property of medicine base starting material and mix by following mass percent: dogwood 2400g, Gynostemma pentaphylla 2000g, Momordica grosvenori 2000g, Ramulus euonymi 500g, balsam pear 1000g, Polyhachis vicina Roger 100g.After above-mentioned raw materials is crushed to 1～2mm, add rice 2000g.
(b) with the mixture of the step a gained container of packing into, add the running water of pH7.5 by 1: 1 weight ratio, in 127 ℃ of sterilization 2h, be cooled to below 30 ℃ after, obtain property of medicine matrix.
(c) under aseptic condition, in 4000g property of medicine matrix, insert the Inonotus obliquus bacterial strain; In 2000g property of medicine matrix, insert Cordyceps strain; In 2000g property of medicine matrix, insert the yellow bacteria strain of mulberry; In 1000g property of medicine matrix, insert Grifola frondosa strain; In 1000g property of medicine matrix, insert the rainbow conk bacterial strain; The inoculative proportion of above-mentioned bacterial classification is 10% of the property of medicine matrix weight inoculated.
(d) connect rearmounted 25 ± 1 ℃ of fermentations of bacterium 45 days.
(e) collect all medicinal fungal substances that steps d obtains, dry to water content below 5% through 40～50 ℃, mix, obtain the medicinal fungal substance goods after nano cell broken wall cracker is crushed to below the 100nm.
(2) finished product detects
According to appendix VID of Pharmacopoeia of People's Republic of China version in 2005, high performance liquid chromatography is measured cordycepin and is reached 1.3mg/g; Measuring total polysaccharides according to UV-VIS spectrophotometry (appendix VA of Chinese Pharmacopoeia version in 2005) is 11%; According to chromium Determination on content method in standard GB/T5009.123-2003 food, measuring biological chromium content is 0.21mg/kg.
Embodiment 4
(1) preparation of medicinal fungal substance
(a) with Chinese herbal medicine after 50～60 ℃ of oven dry, take by weighing dry property of medicine base starting material and mix by following mass percent: dogwood 600g, Gynostemma pentaphylla 600g, Momordica grosvenori 500g, Ramulus euonymi 600g, balsam pear 500g, Polyhachis vicina Roger 200g.Above-mentioned raw materials is crushed to 100 orders.
(b) with the mixture of the step a gained fermentation tank of packing into, add the running water of pH7.5 by 1: 10 weight ratio, in 127 ℃ of sterilizations 40 minutes, be cooled to below 30 ℃ after, obtain property of medicine matrix.
(c) under aseptic condition, according to the 10wt% access Inonotus obliquus bacterial strain of the property of medicine matrix weight of being inoculated.
(d) meet rearmounted 25 ± 1 ℃ of bacterium, tank pressure 0.7～0.8kg/cm
2, throughput 1: 1v/v/ branch, fermented and cultured 8 days.
(e) collect steps d and obtain the Inonotus obliquus medicinal fungal substance,, dry to water content below 5% acquisition Inonotus obliquus medicinal fungal substance through 40～50 ℃ with haplo-effect concentrator vacuum concentration to 1.2.
(f) submerged fermentation method with above-mentioned Inonotus obliquus obtains aweto, the yellow bacterium of mulberry, the corresponding medicinal fungal substance of grifola frondosus, rainbow conk respectively.
(g) get Inonotus obliquus medicinal fungal substance 2000g; Aweto medicinal fungal substance 1500g, the yellow bacterium medicinal fungal substance of mulberry 500g, grifola frondosus medicinal fungal substance 500g, rainbow conk medicinal fungal substance 500g; Mix after nano cell broken wall cracker is crushed to below the 100nm, obtain the medicinal fungal substance goods.
(2) finished product detects
According to appendix VID of Pharmacopoeia of People's Republic of China version in 2005, high performance liquid chromatography is measured cordycepin and is reached 2.5mg/g; Measuring total polysaccharides according to UV-VIS spectrophotometry (an appendix V of Chinese Pharmacopoeia version in 2005 A) is 18.7%; According to chromium Determination on content method in standard GB/T5009.123-2003 food, measuring biological chromium content is 0.43mg/kg.
Embodiment 5
(1) preparation of medicinal fungal substance
(a) with Chinese herbal medicine after 50～60 ℃ of oven dry, take by weighing and mix by following mass percent: dogwood 3000g, Gynostemma pentaphylla 2000g, Momordica grosvenori 1000g, Ramulus euonymi 1500g, balsam pear 1500g, Polyhachis vicina Roger 1000g.With said mixture through three lixiviates: after the running water that adds the pH7.5 of 2 times of mixture weight is for the first time boiled 20 minutes, collect filtrating, the running water that adds the pH7.5 of 2 times of mixture weight was for the second time boiled back 15 minutes; Collect filtrating; The running water that adds the pH7.5 of 2 times of mixture weight was for the third time boiled back 10 minutes, collected filtrating, then three times filtrating was merged; Be concentrated into 6000ml, it is joined in the 5000 gram rice.
(b) with the mixture of the step a gained container of packing into, in 127 ℃ of sterilization 2h, be cooled to below 30 ℃ after, obtain property of medicine matrix.
(c) under aseptic condition, in 500g property of medicine matrix, insert the Inonotus obliquus bacterial strain; In 2000g property of medicine matrix, insert Cordyceps strain; In 500g property of medicine matrix, insert the yellow bacteria strain of mulberry; In 1000g property of medicine matrix, insert Grifola frondosa strain; In 1000g property of medicine matrix, insert the rainbow conk bacterial strain; The inoculative proportion of above-mentioned bacterial classification is 10% of the property of medicine matrix weight inoculated.
(d) connect rearmounted 25 ± 1 ℃ of fermentations of bacterium 45 days.
(e) collect all medicinal fungal substances that steps d obtains, dry to water content below 5% through 40～50 ℃, mix, obtain the medicinal fungal substance goods after nano cell broken wall cracker is crushed to below the 100nm.
(2) intermediate products detect
The polysaccharide of measuring property of medicine matrix according to UV-VIS spectrophotometry (appendix VA of Chinese Pharmacopoeia version in 2005) is 6.7%, and the polysaccharide of Inonotus obliquus fermentation back gained medicinal fungal substance increases to 9.1%; According to chromium Determination on content method in standard GB/T5009.123-2003 food, measure the biological chromium content 0.73mg/kg of property of medicine matrix, the medicinal fungal substance after the grifola frondosus fermentation is brought up to 1.16mg/kg.According to appendix VID of Pharmacopoeia of People's Republic of China version in 2005, high performance liquid chromatography is measured cordycepin content and is brought up to the 9.2mg/g in the Chinese caterpillar fungus mycoplasma by the 0mg/g in its property of medicine matrix.
Explanation thus; The medicinal fungal substance ratio of the two-way fermentation of process just sends out (original) fungi or property of medicine matrix itself or both are added with better drug effect simply mutually; Can produce the physiological function effectiveness of 1+1＞2, can significantly strengthen its active component, improve effects such as drug effect and curative effect.
(3) finished product detects
According to appendix VID of Pharmacopoeia of People's Republic of China version in 2005, high performance liquid chromatography, cordycepin reaches 4.3mg/g; Measuring total polysaccharides according to UV-VIS spectrophotometry (an appendix V of Chinese Pharmacopoeia version in 2005 A) is 20.6%; According to chromium Determination on content method in standard GB/T5009.123-2003 food, measuring biological chromium content is 3.8mg/kg.
Embodiment 6 zooperies
Divide into groups: after the confession examination kunming mice of 54 body weight 18～22g is raised 5 days, be divided into three groups every group 18 male and female half and half at random.Three groups are respectively blank group, hyperglycemia model group, fungi-medicine group.Except the blank group, other two groups of mouse induce hyperglycaemia to reach 14.4mmol/L for three days on end with 200mg/kg dose ejection alloxan.
Dispenser: blank group and hyperglycemia model group mouse were irritated stomach 30 days with continuous normal saline, and the fungi-medicine group was with 25mg/kg dose continuous irrigation stomach 30 days.Respectively organize the eyeball of mouse blood sampling then and measure mice serum glucose with blood sugar test paper (commercially available).
The result: zooperal result of above-mentioned small white mouse such as table 1, fungi-medicine group are compared difference and are had statistical significance (P＜0.05) with model group, fungi-medicine group and compared with normal difference not statistically significant (P＞0.05).This result proves that product according to the invention can significantly reduce in the mouse body, the hyperglycaemia of inducing through alloxan (200mg/kg).
Glucose content in table 1. mice serum (mmol/L) testing result
Claims (9)
1. the raw material with fungi medicinal fungal substance of effect of lowering blood sugar comprises property of medicine matrix and fungi strain, it is characterized in that:
Property of medicine matrix is made up of following raw material: dogwood, Gynostemma pentaphylla, Momordica grosvenori, Ramulus euonymi, balsam pear, Polyhachis vicina Roger, nutrient matrix; Wherein, the source of nutrient matrix is cereal, beans, rice bran, wheat bran;
Fungi strain is made up of following bacterial classification: Inonotus obliquus, Cordyceps sinensis, the yellow bacterium of mulberry, rainbow conk, grifola frondosus;
Wherein, the inoculation weight of fungi strain is 8～12wt% of property of medicine matrix.
2. the raw material of fungi medicinal fungal substance according to claim 1 is characterized in that:
Property of medicine matrix is mixed by the raw material of following drying by mass percentage:
The corresponding relation of each fungi strain and used property of medicine matrix is respectively by mass percentage:
The inoculation weight of fungi strain is 8～12wt% of property of medicine matrix.
3. the raw material of fungi medicinal fungal substance according to claim 2 is characterized in that:
Property of medicine matrix is mixed by the raw material of following drying by mass percentage:
The corresponding relation of each fungi strain and used property of medicine matrix is respectively by mass percentage:
The inoculation weight of fungi strain is the 10wt% of property of medicine matrix.
4. preparation method with fungi medicinal fungal substance of effect of lowering blood sugar is characterized in that: concrete processing step is following:
(a) take by weighing dry property of medicine base starting material and mixing by mass percentage;
(b) in the mixture of step a gained, add the running water of pH7～8 of 1～10 times of mixture quality, in 100～127 ℃ of sterilization 1～10h, obtain property of medicine matrix;
(c) by the corresponding relation of each fungi strain and used property of medicine matrix; The property of medicine matrix of allocation step b gained inserts fungi strain respectively under aseptic condition, inoculative proportion is 8～12wt% of property of medicine matrix; 23～27 ℃ of two-way fermentations 4～45 days, obtain medicinal fungal substance respectively;
(d) collect all medicinal fungal substances that step c obtains, mix obtaining this product after oven dry is pulverized.
5. the preparation method of fungi medicinal fungal substance according to claim 4 is characterized in that, the two-way fermentation mode of employing is solid fermentation or liquid deep layer fermenting.
6. the preparation method of fungi medicinal fungal substance according to claim 5 is characterized in that, the liquid deep layer fermenting condition is: 25～27 ℃ of temperature, tank pressure 0.7～0.8kg/cm
2, throughput 1: 1v/v/ branch.
7. according to the preparation method of claim 4 or 6 described fungi medicinal fungal substances, it is characterized in that the inoculative proportion of said fungi strain is the 10wt% of property of medicine matrix.
8. the fungi medicinal fungal substance for preparing like the said method of claim 7.
9. the application of fungi medicinal fungal substance as claimed in claim 8 in preparation electuary, pill, tablet, capsule.
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