CN102935095B - Bidirectional solid fermentation irpex cacteus mycoplasm extractive and preparation method and application thereof - Google Patents
Bidirectional solid fermentation irpex cacteus mycoplasm extractive and preparation method and application thereof Download PDFInfo
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Abstract
The invention relates to a bidirectional solid fermentation irpex cacteus mycoplasm extractive and a preparation method and application thereof and belongs to the field of medical fungi. Irpex cacteus is subjected to liquid culture with stock culture to obtain seeds at logarithmic phase, the seeds are inoculated to a seed bottle containing a barley culture medium under aseptic condition, and seed bottle fermental cultivation and fermentation bottle fermentation are performed to produce, cultivate and extract. The bidirectional solid fermentation irpex cacteus mycoplasm extractive and the preparation method and application thereof can treat metabolin of glomeruli nephritides and simultaneously change original performances of the barley through growth and metabolism of the irpex cacteus.
Description
Technical field
The invention belongs to field of medicinal fungus, refer in particular to a kind of white rake teeth bacterium mycoplasma extract and preparation method and its usage.
Background technology
White rake teeth bacterium
Irpex lacteusFr. belong to a kind of fungi of Basidiomycetes Aphyllophorales hedgehog fungus section rake teeth Pseudomonas, call white bag hole.This culture presevation is at China Committee for Culture Collection of Microorganisms's common micro-organisms center, and deposit number is: CGMCC No.1016.Its sporophore is born on the dry wood and dead and dying trees of deciduous tree.Morphological specificity: basidioma (sporophore) is calm, expansion, white, soft leather matter; Sometimes fall to give birth to, be multiple watt shape storied, fan-shaped, narrow and small base portion is arranged; The capping oyster white, have down, and spun silk sample gloss, have ring grain, and thin edge is lower curved, wavy to shallow splitting.Bacterial context white.Thorn is irpiciform, short, irregular, and white is to yellow-white, the spore ellipse is level and smooth, colourless, cystidium clavate or fusiform, draped over one's shoulders crystallization sometimes, mainly is distributed in the provinces and regions such as Forest in Changbai Mountain Forest Region and Heilungkiang, Hebei, Shanxi, Gansu, Jiangxi, Fujian, Yunnan and Tibet.
The medicinal wild sporophore of collection among the people that only limits in history of white rake teeth bacterium, its sporophore is born on the dry wood and dead and dying trees of deciduous tree.More difficult collecting, so sporophore is more rare, can't collect in a large number.
Chronic glomerulonephritis is caused by the infringement of multiple primary glomerular disease reason, take proteinuria, blood urine, oedema, the hypertension class disease as common clinical manifestation.Kidney section common disease and frequently-occurring disease.Wherein most patients have kidney function damage in various degree, and are carrying out property and increase the weight of, the gradual renal failure that transfers to.Thereby treating timely and effectively chronic glomerulonephritis is the important topic of researcher.
The eighties-nineties, professor Song Qiyin of Changchun Traditional Chinese Medical College researchs and develops first the method for the white rake teeth bacteria liquid of application fermentation and produces white rake teeth granulose treatment chronic glomerulonephritis, i.e. in the market Chinese patent medicine " Yi-Shen-Kang " capsule.Yi-Shen-Kang capsule, major ingredient are the crude extracts of rake teeth granulose.Content is that brown is to auburn powder; Distinguish the flavor of micro-sour and astringent.Effect with eliminating damp-heat.For the burnt damp-heat syndrome person of chronic glomerulonephritis subordinate.But the patient takes accidental stomach discomfort, dry, it is poor to receive, and feels sick.
The production of medicinal fungi at present generally has liquid fermenting and two kinds of forms of solid fermentation.More application liquid fermenting production both at home and abroad is to use for reference antibiotic production technique, and application of fermentation tank advantage is that higher mechanize and automatization are arranged, and industrial scale is large.But because medicinal fungi self does not generally produce microbiotic, or the microbiotic bacteriostatic action produced a little less than, can not be to good antagonistic actions of other microorganisms such as bacteriums, therefore easy pollution microbes and pouring in down a chimney during the fermentation, cause heavy losses.Its investment of production equipment is huge, consumes energy also more, and production cost is higher.
The solid fermentation of medicinal fungi sees the production of Chinese medicine " Medicated Leaven " and " Pinellia leaven " before 2000 the earliest, is to utilize natural mould to ferment, and its effect is to produce to help digestion and the function of stomach.Its technique is used till today always, but the fermentation of " Medicated Leaven " and " Pinellia leaven " is not pure-blood ferment, the bad control of technique.Along with the development of biotechnology, adopted pure strain inoculation in recent years, the quality supervision system that the science of setting up is controlled, the stability of its fermenting process have obtained solving well.The Zhuan Yi of Nanjing University of Traditional Chinese Medicine teaches in the research eighties in last century with medicinal fungi " Chinese scholartree ear "
Trametes robiniophilaMurr. be fermented bacterium, on nutraceutical matrix, carry out solid fermentation take agricultural byproducts, its medicinal mycoplasma claims " locust tree microellobosporia ".Develop the new drug " Huaier Granule " with antitumor efficacy, propose simultaneously the novel amphicheirality's solid fermentation of medicinal fungi engineering theory.Its principle of the novel amphicheirality's solid fermentation of medicinal fungi engineering (hereinafter to be referred as " two-way fermentation ") is selected specific fermented bacterium, with specific Chinese medicinal materials as property of medicine matrix, use again agricultural byproducts as nutraceutical matrix (being collectively referred to as wholeness matrix), both sides form a fermentation combination altogether, carry out under given conditions two-way fermentation, leavened prod becomes medicinal fungal substance, and it is a ground zero of biotechnology.It can change performance, the enhancing drug effect of former medicine and enlarge indication.Product after fermentation is larger than the drug effect of the independent use of two kinds of materials that ferment, i.e. " 1+1>2 ".
Barley (
Hordeum vulgare) be the seed of Gramineae annual herb plant, be food and medicine, Compendium of Material Medica record: " in the barley flavor is sweet, property puts down, has that dietotherapy is swollen, the long-pending feed that disappears, flat stomach are quenched the thirst, relieved summer heat heat extraction, beneficial controlled atmosphere, wide chest atmosphere, bad, the strong blood vessels of qi-restoratives, beneficial color, real the five internal organs, change the merit of paddy food ".Modern study proves, Fructus Hordei Vulgaris tea contains 17 kinds of trace elements of needed by human body, and 19 kinds with upper amino acid, is rich in multivitamin and unsaturated fatty acids, protein and food fibre.In order to control accumulation of food in the stomach and intes tine due to indigestion, the vexed abdominal distension of gastral cavity, weakness of the spleen and the stomach disease such as have loose bowels.
Summary of the invention
The invention provides a kind of amphicheirality's solid fermentation white rake teeth bacterium mycoplasma extract and preparation method and its usage, cultivate extract and make the patient produce stomach discomfort, dry, receive the problem of the side effect such as poor, nauseating to solve white rake teeth bacteria liquid.
The technical scheme that the present invention takes is: the preparation method of the white rake teeth bacterium of amphicheirality's solid fermentation mycoplasma extract is:
(1) white rake teeth bacterium is planted and carries out the seed that liquid culture obtains logarithmic phase with mother, this seed is inoculated in the seed bottle of barley substratum under aseptic condition;
(2) seed bottle fermentation culture: at 22 ℃-26 ℃ of temperature, atmospheric moisture 65%-70% environment bottom fermentation 7-9 days;
(3) the fermentation flask fermentative production is cultivated: 22 ℃-26 ℃ of temperature, atmospheric moisture 65%-70% environment bottom fermentation 18-21 days, obtain the mycoplasma of white rake teeth bacterium;
(4) this mycoplasma is with 80 ℃ ~ 90 ℃ hot water extraction 2 ~ 3 times, and 3 hours for the first time, 2 hours for the second time, 1 hour for the third time, merging filtrate, surveyed the clear cream of relative density 1.05 ~ 1.10 while being condensed into 60 ℃, and 50 ℃ ~ 55 ℃ drying under reduced pressure, be ground into fine powder.
Barley substratum of the present invention is: the barley corn that is 95%-97% by weight ratio soaks, and makes water content reach 50-55%, adds the terra alba of weight ratio 1%-2%, the sucrose of weight ratio 2%-3%.
The white rake teeth bacterium mycoplasma extract that preparation method by the white rake teeth bacterium of amphicheirality's solid fermentation mycoplasma extract obtains is applied in the burnt damp-heat syndrome person's of preparation treatment chronic glomerulonephritis subordinate medicine.
The present invention utilizes barley to run concurrently and support matrix (title wholeness matrix) as property of medicine matrix, and inoculation has the white rake teeth bacterium for the treatment of glomerulonephritis effect, and both sides form a fermentation combination altogether, carry out under given conditions two-way fermentation, and leavened prod becomes medicinal fungal substance.Barley can provide the nutritive ingredient of white rake teeth bacteria growing, make white rake teeth bacterium mycelial growth breeding, obtain having the brightic metabolite for the treatment of, simultaneously by the metabolism and growth of white rake teeth bacterium, change the original performance of barley, strengthen the function of strengthening the spleen to promote digestion, relax stomach discomfort due to white rake teeth bacterium metabolite, dry, it is poor to receive, and feels sick and waits side effect.
Embodiment
White rake teeth bacterium of the present invention
Irpex lacteusFr, this culture presevation is at China Committee for Culture Collection of Microorganisms's common micro-organisms center, and deposit number is: CGMCC No.1016.
White rake teeth bacterium is planted and carries out liquid culture to obtain the method for seed of logarithmic phase as follows with mother:
(1) primary inclined plane bacterial classification:
Substratum: glucose 2-3%, potato 20-25%, agar 2.2%, MgSO
40.2%, KH
2PO
40.3%, VB
10.05%;
Slant strains culture temperature 25-27 ℃, mycelia was covered with inclined-plane in 120 hours, placed 2-4 ℃ of refrigerator and deposited standby;
(2) secondary shaking flask bacterial classification:
Substratum: glucose 2%, yeast extract 0.5%, KH
2PO
40.2%, NaH
2SO
40.15%, peptone 0.2%, VB
10.01%, pH6.5;
Get the sturdy dense slant strains of mycelia, under aseptic condition, press the inoculation of 1:4 inoculum size, cultivated 4 ~ 6 days in 26 ± 1 ℃ of shaking tables; Result: bacterium liquid is clear and bright, and yellow has dense fruit aroma, and the bacterium ball is radial; Microscopy: methylene blue simple stain mycelia color depth, and sturdy, branch is many, and clamp connexion is obviously visible; This period is the animated period of hyphal development (logarithmic phase) just, again without living contaminants, can transfer solid culture to.
The preparation of embodiment 1 capsule of the present invention
(1) white rake teeth bacterium is planted and carries out the seed that liquid culture obtains logarithmic phase with mother, this seed is inoculated in the seed bottle of barley substratum under aseptic condition; Described barley substratum is: the barley corn that is 95% by weight ratio soaks, and makes water content reach 50%, adds the terra alba of weight ratio 1%, the sucrose of weight ratio 2%;
(2) seed bottle fermentation culture: 26 ℃ of temperature, atmospheric moisture 70% environment bottom fermentation 7 days;
(3) the fermentation flask fermentative production is cultivated: 26 ℃ of temperature, atmospheric moisture 70% environment bottom fermentation 18 days, obtain the mycoplasma of white rake teeth bacterium;
(4) this mycoplasma was with 90 ℃ of hot water extraction 3 times, 3 hours for the first time, 2 hours for the second time, 1 hour for the third time, merging filtrate, the clear cream of survey relative density 1.10 while being condensed into 60 ℃, 55 ℃ of drying under reduced pressure, be ground into fine powder, add the auxiliary material for preparing capsule, prepare according to a conventional method capsule.
The preparation of embodiment 2 tablets of the present invention
(1) white rake teeth bacterium is planted and carries out the seed that liquid culture obtains logarithmic phase with mother, this seed is inoculated in the seed bottle of barley substratum under aseptic condition; Described barley substratum is: the barley corn that is 96% by weight ratio soaks, and makes water content reach 53%, adds the terra alba of weight ratio 1.5%, the sucrose of weight ratio 2.5%;
(2) seed bottle fermentation culture: 22 ℃ of temperature, atmospheric moisture 65% environment bottom fermentation 9 days;
(3) the fermentation flask fermentative production is cultivated: 22 ℃ of temperature, atmospheric moisture 65% environment bottom fermentation 21 days, obtain the mycoplasma of white rake teeth bacterium;
(4) this mycoplasma is with 80 ℃ of hot water extraction 2 times, and 3 hours for the first time, 2 hours for the second time, merging filtrate, surveyed the clear cream of relative density 1.05 while being condensed into 60 ℃, and 50 ℃ of drying under reduced pressure, be ground into fine powder, adds the auxiliary material for preparing tablet, prepares according to a conventional method tablet.
The preparation of embodiment 3 granules of the present invention
(1) white rake teeth bacterium is planted and carries out the seed that liquid culture obtains logarithmic phase with mother, this seed is inoculated in the seed bottle of barley substratum under aseptic condition; Described barley substratum is: the barley corn that is 97% by weight ratio soaks, and makes water content reach 55%, adds the terra alba of weight ratio 2%, the sucrose of weight ratio 3%;
(2) seed bottle fermentation culture: 24 ℃ of temperature, atmospheric moisture 67% environment bottom fermentation 8 days;
(3) the fermentation flask fermentative production is cultivated: 24 ℃ of temperature, atmospheric moisture 67% environment bottom fermentation 20 days, obtain the mycoplasma of white rake teeth bacterium;
(4) this mycoplasma was with 85 ℃ of hot water extraction 3 times, 3 hours for the first time, 2 hours for the second time, 1 hour for the third time, merging filtrate, the clear cream of survey relative density 1.08 while being condensed into 60 ℃, 52 ℃ of drying under reduced pressure, be ground into fine powder, add the auxiliary material for preparing granule, prepare according to a conventional method granule.
The curative effect report of the white rake teeth fungus extract of solid fermentation of the present invention preparation for treating chronic glomerulonephritis
Clinical dataAll have a chronic renal glomerulonephritis medical history, between 24h quantity of proteinuria 0. 3～3g, Scr<350umo/ L person classifies research object as, observe altogether 264 examples, inpatient's 120 examples wherein, outpatient's 144 examples, according to its medical and the order of being in hospital, be divided at random treatment group 132 examples, control group 132 examples.Exclusion standard: (1) non-gravid woman, non-allergic constitution and to this medicine without allergy sufferers; (2) the serious primary disease person such as cardiovascular, the liver of nonjoinder, hemopoietic system; (3) other than psychotic patient; (4) person that adheres to the medication treatment.This problem is by above-mentioned Standard Selection 264 examples, treatment group 132 examples, and the male sex's 64 examples wherein, women's 68 examples, the age, the course of disease was between April～8 year between 14～64 years old.Merge creatinine, abnormal person's 56 examples of blood urea nitrogen; Hemochrome reduction person 62 examples; Hyperpietic's 33 examples.Control group 132 examples, the male sex's 62 examples wherein, women's 70 examples, the age, the course of disease was between 6 months～7 years between 15～68 years old.Merge creatinine, abnormal person's 44 examples of blood urea nitrogen; Hemochrome reduction person 50 examples; Hyperpietic's 30 examples.Two groups of cases are statistically without significant difference.
Methods for the treatment of
Primary Care: (1) infection control.(2) control blood pressure.(3) there is oxypathy person to give the correction oxypathy.
Treatment group: take the capsule that the white rake teeth bacterium of amphicheirality's solid fermentation mycoplasma extract is made, each 2, every day 3 times, 30 days is 1 course for the treatment of, treats and adds up curative effect 3 courses for the treatment of.
Control group: taking the liquid fermenting preparation is each 2 of Yi-Shen-Kang capsule (production of Changchun Yi-Shen-Kang Biology Pharmacy Co., Ltd), and every day 3 times, 30 days is 1 course for the treatment of, treats and adds up curative effect 3 courses for the treatment of.
Observation index: cardinal symptom changes: comprise soreness of the waist and knees, and fatigue and weakness, oedema, dizzy etc.Situation appears in side effect: comprise stomach discomfort, and dry, it is poor to receive, and feels sick etc.Lab index: do the inspections such as 24h quantity of proteinuria, serum creatinine, blood urea nitrogen, oxyphorase before and after treatment.
Statistical procedures: measurement data adopts the T check, and enumeration data adopts x
2Check.
The criterion of therapeutical effect recovery from illness: symptom and the sign completely dissolves such as oedema, urine protein examination continues negative, or the 24h quantity of proteinuria continues to be less than 200mg, urine erythrocyte disappearance under high power lens.The arena counting is normal, and graft function is normal.Produce effects: the symptom such as oedema and sign disappear substantially, and the urine protein examination continuous decrease is more than 50%, and under high power lens, urine erythrocyte is no more than 3, and the arena counting approaches normal.Normal renal function or normal, differ and be no more than 15% with normal value.Effectively: the symptom such as oedema and sign are clearly better, 1 of urine protein examination continuous decrease+or 24h quantity of proteinuria continuous decrease more than 25%, and under high power lens, urine erythrocyte is no more than 5, normal renal function or be improved.Invalid: clinical manifestation and above-mentioned laboratory examination all are not improved or add severe one.
Treatment result
1, curative effect this group 264 examples relatively between two groups, treatment group: 132 examples, recovery from illness 16 examples, produce effects 44 examples, effective 66 examples, invalid 6 examples, total effective rate 95.4%.Control group: 132 examples, recovery from illness 14 examples, produce effects 40 examples, effective 69 examples, invalid 9 examples, total effective rate 93. 1%.Two groups of total effective rate comparison P values are greater than 0. 05, and difference is not very significant.
2, between two groups, treatment front and back lab index changing conditions is shown in (table one)
Before and after table 2 liang group patient treatment, lab index changes
Annotate: with compare * P<0. 01 * * P<0. 05 and treatment of control group before this group is treated after compare △ P>0. 05
3 routine stomach discomforts appear after the capsule that 3, side effect treatment group is made with solid fermentation, 6 routine drys, and it is poor that receiving appears in 2 examples, the 0 example symptom of feeling sick.The capsule that the control group liquid fermenting is made (Yi-Shen-Kang) 40 routine stomach discomforts, 32 routine drys, it is poor that receiving appears in 20 examples, the nauseating symptom of 12 example.
The white rake teeth bacterium of conclusion amphicheirality's solid fermentation mycoplasma extract and liquid fermentation production preparation all have curative effect preferably to glomerulonephritis, but the liquid fermentation production side effect is more, and the side effect of the white rake teeth bacterium of amphicheirality's solid fermentation mycoplasma extract is less, so the white rake teeth bacterium of amphicheirality's solid fermentation mycoplasma extract formulation is better than the liquid fermenting preparation.
Claims (5)
1. the white rake teeth bacterium of amphicheirality's solid fermentation mycoplasma extract is characterized in that it is obtained by following method:
(1) white rake teeth bacterium is planted and carries out the seed that liquid culture obtains logarithmic phase with mother, this seed is inoculated in the seed bottle of barley substratum under aseptic condition;
(2) seed bottle fermentation culture: at 22 ℃-26 ℃ of temperature, atmospheric moisture 65%-70% environment bottom fermentation 7-9 days;
(3) the fermentation flask fermentative production is cultivated: 22 ℃-26 ℃ of temperature, atmospheric moisture 65%-70% environment bottom fermentation 18-21 days, obtain the mycoplasma of white rake teeth bacterium;
(4) this mycoplasma is with 80 ℃ ~ 90 ℃ hot water extraction 2 ~ 3 times, and 3 hours for the first time, 2 hours for the second time, 1 hour for the third time, merging filtrate, surveyed the clear cream of relative density 1.05 ~ 1.10 while being condensed into 60 ℃, and 50 ℃ ~ 55 ℃ drying under reduced pressure, be ground into fine powder.
2. the white rake teeth bacterium of amphicheirality's solid fermentation according to claim 1 mycoplasma extract, it is characterized in that: described barley substratum is: the barley corn that is 95%-97% by weight ratio soaks, make water content reach 50-55%, add the terra alba of weight ratio 1%-2%, the sucrose of weight ratio 2%-3%.
The white rake teeth bacterium of amphicheirality's solid fermentation as claimed in claim 1 mycoplasma extract the preparation method, comprise the following steps:
(1) white rake teeth bacterium is planted and carries out the seed that liquid culture obtains logarithmic phase with mother, this seed is inoculated in the seed bottle of barley substratum under aseptic condition;
(2) seed bottle fermentation culture: at 22 ℃-26 ℃ of temperature, atmospheric moisture 65%-70% environment bottom fermentation 7-9 days;
(3) the fermentation flask fermentative production is cultivated: 22 ℃-26 ℃ of temperature, atmospheric moisture 65%-70% environment bottom fermentation 18-21 days, obtain the mycoplasma of white rake teeth bacterium;
(4) this mycoplasma is with 80 ℃ ~ 90 ℃ hot water extraction 2 ~ 3 times, and 3 hours for the first time, 2 hours for the second time, 1 hour for the third time, merging filtrate, surveyed the clear cream of relative density 1.05 ~ 1.10 while being condensed into 60 ℃, and 50 ℃ ~ 55 ℃ drying under reduced pressure, be ground into fine powder.
4. the preparation method of the white rake teeth bacterium of amphicheirality's solid fermentation according to claim 3 mycoplasma extract, it is characterized in that: described barley substratum is: the barley corn that is 95%-97% by weight ratio soaks, make water content reach 50-55%, the terra alba that adds weight ratio 1%-2%, the sucrose of weight ratio 2%-3%.
5. the white rake teeth bacterium of amphicheirality's solid fermentation as claimed in claim 1 mycoplasma extract is applied in the burnt damp-heat syndrome person's of preparation treatment chronic glomerulonephritis subordinate medicine.
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| CN110804557A (en) * | 2016-08-31 | 2020-02-18 | 郭清子 | Culture method and culture medium of irpex cacteus |
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Address after: 130012 No. 767, fire road, Qianjin Street, Jilin, Changchun Patentee after: Changchun Pingan xingyishenkang Biological Pharmaceutical Co., Ltd Address before: 130012 No. 767, fire road, Qianjin Street, Jilin, Changchun Patentee before: Changchun Yishenkang Biopharmaceutical Co., Ltd. |