CN110527702A - Utilize the method for mutagenic obtained Inonotus obliquus bacterial strain production Fuscoporia obliqua polysaccharide - Google Patents
Utilize the method for mutagenic obtained Inonotus obliquus bacterial strain production Fuscoporia obliqua polysaccharide Download PDFInfo
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Abstract
The invention discloses the methods using mutagenic obtained Inonotus obliquus bacterial strain production Fuscoporia obliqua polysaccharide, are related to food microorganisms applied technical field.Inonotus obliquus new strains ferment in the liquid culture medium through examining the rice bran wheat bran complete feed for meeting contaminants in foods in the method, it after the fermentation material for producing Inonotus obliquus mycelium raw material and fermentation liquid, washed by multiple frequency ultrasonic assisted extraction, freeze concentration, citric acid extraction displacement heavy metal ion, alcohol precipitation alcohol, be lyophilized and obtain the Fuscoporia obliqua polysaccharide product of qualification.The rice bran wheat bran of low value is converted to the Fuscoporia obliqua polysaccharide with plurality of health care functions of high level by the present invention; to reduction production cost, the reasonable higher value application of low side resource high-efficiency, protection health of masses; all there is beneficial society and economic effect, thus there is practicability.
Description
Technical field
The present invention relates to food microorganisms applied technical fields more particularly to a kind of new using mutagenic obtained Inonotus obliquus
The method that bacterial strain produces Fuscoporia obliqua polysaccharide, Inonotus obliquus new strains are meeting contaminants in foods through examining in the method
Rice bran wheat bran complete feed liquid culture medium in ferment, produce the fermentation material of Inonotus obliquus mycelium raw material and fermentation liquid
Afterwards, it washed by multiple frequency ultrasonic assisted extraction, freeze concentration, citric acid extraction displacement heavy metal ion, alcohol precipitation alcohol, be lyophilized and closed
The Fuscoporia obliqua polysaccharide product of lattice.
Background technique
Many dual-purpose of drug and food fungies or medicinal fungi have long usage history in China and widely use crowd.It is existing
Generation science discloses, the mycelium of existing frequently-used rare fungi such as Hericium erinaceus, ganoderma lucidum, grifola frondosus, Inonotus obliquus, Inonotus obliquus etc.,
Amino acid, protein, vitamin, polysaccharide, ucleosides, terpenoid, flavonoids etc. can be generated in fructification or spore
A variety of active ingredients or enrichment various trace elements such as selenium, zinc etc. have and improve the immunity of the human body, is antitumor, enhancing liver function
The multiple efficacies such as energy, anti-oxidant.Just because of this, the interest that international and domestic all kinds of personnel research and develop this kind of fungi of medicine is dense
Thickness, in field of medicaments, field of food, Traditional Chinese health field, agricultural and field of industrial production all there are many researcher and developer,
Obtain numerous research achievement or product, such as ganoderma lucidum capsule, grifola frondosus capsule and Paecilomyces hepiali chen capsule, their economy
It is worth higher.Hericium erinaceus being added in biscuit, Hericium erinaceus biscuit being then made, having become ordinary food, supermarket carries out pin at home
It sells.New Zealand, Japan, the U.S., South Korea etc., which produce, in the world similar rare fungi health food or pharmaceutical products.
For Inonotus obliquus, for its research shows that: Inonotus obliquus antitumor, anticancer, it is anti-oxidant, hypoglycemic,
Strengthen immunity, treatment cardiovascular disease etc. all have certain bioactivity, and medical value with higher is (with reference to text
It offers: 1. Liuping Fan, Shaodong Ding, Lianzhong Ai, et al.Antitumor and
immunomodulatory activity of water-soluble polysaccharide from Inonotus
Obliquus [J] .Carbohydrate Polymers, 2012,90 (2): 870-874；2. Lishuai Ma, Haixia
Chen, Peng Dong, Xueming Lu.Anti-inflammatory and anticancer activities of
Extracts and compounds from the mushroom Inonotus obliquus [J] .Food Chemistry,
2013,139 (1-4): 503-508；3. Xu Lixuan, Cai Jianxiu, Yue Zhiling Inonotus obliquus ethanol extract resist to inside and outside Mice Body
Oxidation research [J] Chinese food journal, 2012,12 (6): 48-54；4. the gorgeous Inonotus obliquus deep fermentation of Wang Hong and
The Hangzhou Primary Study [D] of bacterium powder hypoglycemic drug effect: Zhejiang University, 2013；5. Kim Yong Ook, Han Sang Bae,
Lee Hong Woen, et al.Immunostimulating effect of the endo-polysaccharide
Produced by submerged culture of Inonotus obliquus [J] .Life Sciences, 2005,77:
2438-2456)。
Inonotus obliquus (Inonotus obliquus) also known as inonotus obliquus belong to Basidiomycotina, Hymenomycetes, non-brown Zoopagales, more
Hole Cordycepps, brown transverse hole fungus category, are a kind of large-scale medicinal fungis parasitized on the tree such as birch, elm, and fructification shows class
It is similar to the block-shape morphology of carbon.The fructification surface of Inonotus obliquus is very hard, and color is in yellowish-brown to black etc., and there are many significant
Rugged cracking fritter.Fructification initial stage in tree subcutaneous growth, highlights after decortication, and tiling expansion, volume differs in size,
Color is white at the beginning, slowly becomes yellow, eventually becomes brown, the long 2~8mm of tube, the front end cracking of usual tube.Bacterium
Meat part forms i.e. bristle by two kinds of mycelia and strengthens mycelia and generative hyphae, there is tiny and indistinct bad shape striped.Inonotus obliquus exists
40 °~50 ° of Northern Hemisphere north latitude of area is distributed, such as the states such as Finland, Russia, Japan, Korea.China Inonotus obliquus master
It is distributed in the ground such as Jilin, Heilungkiang, Tibet and the Inner Mongol.The Inonotus obliquus speed of growth under wild state is very slow, because
Growing into the more than ten years on birch for it just possesses significant medical value, so, wild Inonotus obliquus is expensive, about
It is 7~8 times of Phellinus price.Inonotus obliquus is currently under in environment in imminent danger, and priority protection coefficient is higher, is listed in one
Grade protection class ranks.As it can be seen that since wild Inonotus obliquus resource is very rare, there is the money that must solve Inonotus obliquus
Source scarcity problem.The acquisition modes of present Inonotus obliquus are mainly artificial cultivation, are used as medicine with fructification or conidia powder, but Planting
Train that the Inonotus obliquus period is long, production efficiency is not high, large labor intensity, limited by season, environment etc., be subject to pest and disease damage, quality with
Yield is unstable.Rare fungi fermentation technology can overcome the shortcomings of traditional fructification cultivation, using liquid or solid state fermentation
Method production Inonotus obliquus fermentation material is simultaneously subject to the thinking of development that deep processing application meets innovation driving, has good prospect.
The application that Inonotus obliquus all will be further pushed to the progress that the research and development of Inonotus obliquus fermentation material obtains, brings actual
Social and economic benefit.Just because of this, just it is necessary to study the productions of new Inonotus obliquus fermentation material and its deep processed product
Method, such as consider the birch that some low side processing of farm products byproduct sources such as wheat bran, rice bran Efficient Conversion are high value is brown
Pore fungi fermentation raw material, it is expected to reduce the production cost of Inonotus obliquus fermentation material to reduce finished product such as Inonotus obliquus class health care
The price of food or drug, interest concessions promote health of masses, this is also one of the invention patent focus in ordinary people.
China is the big producer of rice and wheat, and rice bran and wheat bran are resourceful, and annual output rice bran is about 10,000,000 tons,
Wheat bran is about 30,000,000 tons.Wheat bran refers both to wheat bran hereinafter.Rice bran and wheat bran are as paddy or the by-product of wheat processing, nutrition
Ingredient is abundant, cheap.Contain in rice bran and enriches good protein, active polysaccharide, fat and tocotrienols, tocopherol
Etc. the significant active material of physiological functions, wheat bran rich in the nutritional ingredients such as fat, protein, minerals, vitamin and cellulose,
Wherein the content of cellulose is up to 18% or more of wheat bran total amount, so, rice bran and wheat bran can grow provide and fill for Inonotus obliquus
Carbon source, the nitrogen source, vitamin and mineral of foot.Under the action of Inonotus obliquus cellulase and other enzyme systems, Inonotus obliquus can
Rice bran and wheat bran are converted to needed nutrient matter and carry out growth metabolism, synthesizes Inonotus obliquus mycelium and types of functionality object
Matter.Therefore, it carries out Inonotus obliquus liquid state fermentation as whole carbon source and nitrogen source with agricultural and sideline product rice bran, wheat bran and produces birch
Simultaneously deep processing has technical feasibility at health food or drug to brown pore fungi fermentation material, and can bring preferable society and economy
Benefit.
Most edible mushroom Liquid-state fermentation production methods use glucose, grain class raw material such as starch, potato or soybean powder
Deng be used as culture medium, even if agricultural and sideline product such as rice bran, wheat bran etc. can be used, and as auxiliary element be added.Original
Inonotus obliquus upgrowth situation on the rice bran wheat bran complete feed liquid culture medium for not adding glucose or other grain class raw materials is not
To be best, rice bran, residual containing some harmful substances such as lead, arsenic, aflatoxin, vomitoxin, pesticide in bran feedstock in addition
It stays object etc. to influence final products quality, therefore the fermentation wheat bran rice bran liquid complete feed to Inonotus obliquus is needed to produce Inonotus obliquus hair
The method of ferment object carries out innovation research.
The liquid of grifola frondosus, ganoderma lucidum, cordyceps sinensis etc. is during the last ten years sent out by the research group of inventor herein Liu Weimin
Ferment has carried out a large amount of research, successively forms multiple achievements such as Master's thesis and patent of invention.The Master's thesis of Liu Weimin guidance
Have: (1) Yang Suohua, frondosa fermentation rice bran prepare polysaccharide (2006)；(2) Gu Huimin, grifola frondosus liquid in rice bran culture medium
State culture produces polysaccharide and the research (2009) for being enriched with organic selenium；(3) it opens and builds, physical method mutagenesis grifola frondosus liquid fermentation rice bran bran
The research (2010) of skin production polysaccharide；(4) Guo Chunmei, induction mutation of bacterium, the liquid fermentation rice bran wheat bran of grifola frondosus produce polysaccharide and richness
Selenium studies (2011)；(5) Li Yanan, grifola frondosus induction mutation of bacterium and fermentation and performance study (2013)；(6) Liu Lili is high
The Ganoderma lucidum submerged fermentation and induction mutation of bacterium (2012) of value conversion rice bran wheat bran；(7) Guo Tianlong, lucidum strain mutagenesis and liquid hair
The full rice bran wheat bran research (2013) of the high-valued conversion of ferment；(8) Chen Jing, cordyceps sinensis liquid fermentation higher value application rice bran wheat bran
Produce the research (2014) of polysaccharide；(9) Zhang Xiaofei, ganoderma strain (Ganoderma lucidum CFCC6043) liquid fermentation are complete
The research (2014) of rice bran wheat bran and active polysaccharide；(10) Cao Xun, grifola frondosus mutagenesis and new and old strain liquid ferment rice bran bran
Skin studies (2015)；(11) Jin Lei, ganoderma lucidum mutagenesis and new and old strain liquid ferment rice bran and wheat bran research (2015)；(12)
Wang Xuemei, with the comparative studies (2016) of Cordyceps strain liquid fermentation rice bran wheat bran complete feed；(13) Li Tingting, worm summer in winter
The research (2017) of careless strain mutagenesis and the new culture medium of new strains liquid state fermentation rice bran wheat bran complete feed.Liu Weimin is as inventor
Patent of invention have: (1) using rice bran wheat bran compound material and grifola frondosus mutagenic strain production polysaccharide method,
20101010579048.4；(2) Grifola frondosa strain for rice bran and wheat bran compound material production polysaccharide,
201010579078.5；(3) bacterial strain for ferment rice bran and wheat bran extracting solution production grifolan,
201110150888.3；(4) bacterial strain for rice bran and wheat bran complete feed liquid state fermentation production grifolan,
201310274913.8；(5) a kind of method of ganoderma lucidum mutant strain liquid state fermentation rice bran wheat bran complete feed production polysaccharide,
201310275061.4；(6) a kind of method of liquid state fermentation rice bran wheat bran complete feed production Chinese caterpillar fungus polysaccharide,
201310274914.2；(7) a kind of method of grifola frondosus mutagenic fungi liquid state fermentation rice bran wheat bran complete feed production polysaccharide,
201310274911.9；(8) bacterial strain for rice bran and wheat bran complete feed liquid state fermentation production ganoderma lucidum polysaccharide,
201310274915.7；(9) production method of Se-rich lucid ganoderma mycelia raw material, 201510996120.6；(10) one plants of production ash trees
Spend mycelial grifola frondosus mutagenic strain, 201510990344.6；(11) the cordyceps sinensis mutagenesis of aweto mycelium is produced
Bacterial strain, 201510996118.9 etc..As seen from the above description, inventor herein Liu Weimin all around trans-utilization rice bran and
Wheat bran efficiently studied by this special topic of high-valued production Rare edible fungus fermentation material, realizes the imagination of innovation and creation, and constantly
Improve the production method of various rare fungal fermentates.
The paper of the Master degree candidate of inventor Liu Weimin research institute guidance and invention relevant to the invention patent in the past
Patent consults the unanswerable problem of related ends when institute in conjunction with enterprise, has been found that Industrialization to be moved towards, relevant
There are still many problems needs to research and solve and be innovated for patented technology and research paper.These problems include different rare true
The acquisition of the efficient new strains of bacterium, the formulation of rice bran bran feedstock standard, the innovation of rare fungal fermentate fermentation process, rare fungi
Fermentation material isolates and purifies new method use and these need to innovate the combination etc. of object.Patent of invention is in design to awarding
It gives uniqueness necessary to patent for invention, obtain substantive distinguishing features outstanding and the innovation and practicality of marked improvement will be to
Give abundant consideration.So the combined innovation of above-mentioned these problems, which should be ensured that relevant patent of invention reaches, authorizes invention specially
Uniqueness, innovation and practicality necessary to economic rights.
Inventor is based on the above considerations and basic to the early-stage study of Inonotus obliquus fermentation material special producing method, mentions
The Inonotus obliquus new strains that the invention patent obtains ferment rice bran wheat bran production Inonotus obliquus fermentation material using ultraviolet mutagenesis are gone out
Application because Inonotus obliquus is a kind of ganoderma lucidum studied different from inventor, grifola frondosus, the dual-purpose of drug and food of cordyceps sinensis
It is complete to obtain Inonotus obliquus new strains high efficiency liquid ferment rice bran wheat bran by the method for ultraviolet mutagenesis before the application for fungi
Also someone had not studied the method for material production Inonotus obliquus fermentation material.
In conclusion field of the present invention is that be used to ferment by Inonotus obliquus wheat bran and rice bran complete feed produces the brown hole of birch
Bacterium fermentation material simultaneously becomes qualified products after deep processing is handled.The invention solves one of critical issue to need by luring
The method of change promotes the fermentability of original strain, obtains a kind of Inonotus obliquus new strains having not been reported, and can be to this new bacterium
Strain carries out gene sequencing, it is ensured that strain used is correct.The invention solves critical issue two to make wheat bran raw rice bran
Standard, because wheat bran and rice bran source are complicated, wherein pollutant kind and its level are different, inventor's previous studies and patent
Do not give enough concerns in this regard, brings indefinite consequence to subsequent product exploitation.The invention solves critical issue it
Three production methods to become qualified products after fermentation material deep processing processing.Above three promise the invention patent and other
All kinds of achievements of inventor and the present inventor Liu Weimin research team are different, unique, innovation and practicality.
The reason of proposing for the critical issue in terms of above three is further explained.It is proposed the invention solves
One of critical issue proposes the method needed through mutagenesis, promote the fermentability of original Inonotus obliquus strain, obtains one
The Inonotus obliquus new strains that kind has not been reported, and gene sequencing can be carried out to this new strains, it is ensured that strain used is correct.This is asked
The background of topic is that the previous research achievement of inventor team is related to the Rare edible fungus such as ganoderma lucidum, grifola frondosus, cordyceps sinensis mostly,
Just tentatively it is related to Inonotus obliquus original strain fermentation wheat bran before, but original strain fermentation wheat bran is inefficient.In view of front
The reason of pharmacology for the Inonotus obliquus introduced discloses and functional components are abundant and resource scarcity price is more expensive etc., so
Consider the method for research and utilization Inonotus obliquus new strains fermentation wheat bran rice bran production Inonotus obliquus fermentation material.According to inventor with
Toward research experience, this kind of rare fungi fermentation wheat bran rice bran is firstly the need of the adaptability problem for solving strain.It would therefore be desirable to
Its fermenting property is improved by the method for induction mutation of bacterium.In addition in the course of the research, through a large amount of strain processing step
After to guarantee that mistake does not occur in strain, it should carry out gene sequencing.
It is proposed the invention solves critical issue two be make wheat bran raw rice bran standard background be it is former we
Research and patent do not give enough concerns in this regard, to subsequent product exploitation bring unexpected consequence sometimes.With
Preceding we always wish that studied achievement is capable of handling various wheat brans and rice bran, therefore relax use intentionally to wheat bran raw rice bran
Limitation does not carry out raw material standard formulation, but this have the consequence that occurs not in end product quality characterization in research model
Interior some exceptional values, such as certain determining heavy metals result excessive problems are enclosed, tracing it to its cause is because wheat bran and raw rice bran are come
Source is complicated, and quality is difficult to keep similar level, and unknown pollutant eventually enters into end product.It is directed to certain specific dirts originally in this way
The invention production method for contaminating object and its possible content and designing, it is possible to occur other pollutants in the product, so to weigh
The production method of new design fermentation material, copes with the new pollutant being likely to occur.In this way, the control of wheat bran and raw rice bran quality
Just become the problem of new invention Patent design must be taken into consideration with standard formulation problem.The present invention proposes making for raw material wheat bran and rice bran
Use standard.
It is proposed the invention solves critical issue three for fermentation material deep processing processing after become qualified products life
The background of production method is that the reasonable conjunction of entire fermentation material production method should be considered on the basis of the first two critical issue solves
Rationality.Since wheat bran raw rice bran is possible to bring unknown pollutant into, consider that addition enrichment mortifier may not be just in the medium
It can achieve the purpose that remove all contaminants, such hypha fermentation product is such as used as final products application, wherein being still likely to occur
Unknown pollutant and content is exceeded, so the reasonable processing method of the case where being likely to occur for this should be after fermentation to hair
Ferment product designs extraction separation method, obtains qualified final product.Based on this understanding, the invention patent, which is designed, is more closed
The fermentation material production and processing method of reason.
The present invention needs to obtain Inonotus obliquus new strains by the method for mutagenesis.The selection of microorganism mainly has physics
Mutagenesis, chemical mutagenesis, genetic recombination etc., the present invention be avoid using poisonous and hazardous chemical mutagen and field of food also not
The gene engineering method approved by masses will utilize relatively easy easy ultraviolet mutagenesis technology, starting strain made to be in mutagenesis
Extreme environment, expand the site range of mutation to greatest extent, a possibility that raising obtains direct mutation bacterial strain.The present invention is using purple
This common method of mutagenesis of outer mutagenesis, although the method does not have novelty, the bacterial strain performance that the method obtains is had occurred
Innovative variation, the unique, innovation and practicality when fermenting wheat bran rice bran complete feed culture medium.
Inventor mainly solves the critical new problem of above three, constructs the contents of the present invention.
The ganoderma lucidum new strains of gained fermentation wheat bran rice bran complete feed liquid culture medium of the invention are that invention obtains for the first time.
Wheat bran of the invention is to execute the former Ministry of Agriculture to eat feed wheat bran standard NY/T 3218-2018 using standard, outside
Add the level of pollution not more than 3mg/kg of Heavy Metals lead, arsenic, mercury, cadmium, chromium, applying pesticides accord with during wheat planting
Agrotechnical specification is closed, is done harm in wheat growth without serious head blight, harvesting season causes wheat rotten without continuous rainwater, small Meccah
Wheat bran storage and transportation not will lead to mildew after work.Rice bran is to execute the former agricultural additional main weight of ministerial standard NY/T122-1989 using standard
Metallic lead, arsenic, mercury, cadmium, chromium level of pollution limit the quantity not more than 3mg/kg, applying pesticides meet agriculture during Rice Cropping
Skill specification, paddy growth is in the process without serious plant disease, and harvesting season causes paddy rotten without continuous rainwater, rice bran after paddy processing
Storage and transportation not will lead to mildew.
The present invention provides above-mentioned Inonotus obliquus new strains in the rice bran wheat bran complete feed for meeting contaminants in foods through examining
Liquid culture medium in ferment, the method for preparing Fuscoporia obliqua polysaccharide, concrete composition, fermentation condition including culture medium,
The measured value of extraction and impurity-removing method, mycelia and polysaccharide yield and the polysaccharide index of quality.Inonotus obliquus total starches obtained
Mycotoxin aflatoxin B1, the limitation of B2 and vomitoxin, heavy metal lead, arsenic, mercury, cadmium, the limitation of chromium and the more bacterium of pesticide
Spirit, the residue limits process measurement of avermectin, butachlor are not higher than GB2761-2017, GB2762-2017, GB2763-
Limited Doses as defined in 2016 are not detected.
Summary of the invention
The present invention is a kind of method for producing Fuscoporia obliqua polysaccharide using mutagenic obtained Inonotus obliquus new strains, in the party
Inonotus obliquus new strains carry out in the liquid culture medium through examining the rice bran wheat bran complete feed for meeting contaminants in foods in method
It ferments, it is dense by multiple frequency ultrasonic assisted extraction, freezing after the fermentation material for producing Inonotus obliquus mycelium raw material and fermentation liquid
Contracting, citric acid extraction displacement heavy metal ion, alcohol precipitation alcohol wash, be lyophilized and obtains the ganoderma lucidum polysaccharide product of qualification.
The technical solution used in the present invention is as follows: utilizing a kind of Inonotus obliquus new strains obtained through ultraviolet mutagenesis, In
It ferments in liquid culture medium through examining the rice bran wheat bran complete feed for meeting contaminants in foods, produces Inonotus obliquus bacterium
After the fermentation material of filament raw material and fermentation liquid, by multiple frequency ultrasonic assisted extraction, freeze concentration, citric acid extraction displacement heavy metal
Ion, alcohol precipitation alcohol washes, is lyophilized and obtains qualified Fuscoporia obliqua polysaccharide product.
In one aspect of the invention, a kind of to produce Fuscoporia obliqua polysaccharide using mutagenic obtained Inonotus obliquus new strains
Method, as steps described below carry out:
(1) examine wheat bran and rice bran, Heavy Metals lead, arsenic, mercury, cadmium, chromium level of pollution not more than 3mg/kg,
Remaining corresponds with the former Ministry of Agriculture and eats feed wheat bran standard NY/T 3218-2018 and rice bran standard NY/T122-1989, small
Applying pesticides meet agrotechnical specification during wheat seeds are planted, and do harm in wheat growth without serious head blight, gather in season without continuous
Rainwater causes wheat rotten, and wheat bran storage and transportation not will lead to mildew after wheat processing, and applying pesticides meet agriculture during Rice Cropping
Skill specification, paddy growth is in the process without serious plant disease, and harvesting season causes paddy rotten without continuous rainwater, rice bran after paddy processing
Storage and transportation not will lead to mildew；
(2) rice bran and wheat bran are added into 121 DEG C of sterilizing 30min of the desired amount of water；
(3) fermentation medium of Inonotus obliquus new strains, rice are directly used in using rice bran and wheat bran complete feed as carbon source, nitrogen source
Chaff concentration is 0.1-10g/100mL, and wheat bran concentration is 0.1-10g/100mL, and the total concentration of rice bran and wheat bran is up to 10.1g/
100mL, add potassium dihydrogen phosphate 0.015-0.25g/100mL, epsom salt 0.015-0.25g/100mL, pH naturally,
250mL shaking flask charging 100mL, fermentor filling rate 75%, inoculum concentration 10%, 24-28 DEG C of cultivation temperature, shaking speed 150r/
Min or fermentation rotating speed of agitator 50-150r/min, fermentation time 5-12d；
(4) liquid culture is centrifugally separating to obtain mycelium and fermentation liquid, and measures the production of exocellular polysaccharide in fermentation liquid
Amount；
(5) the ultrasonic wave added liquid circulation of 3.5kHz and 1.5kW is added to extract 1.5h 60 DEG C of hot water of gained mycelium, from
Heart separation takes clear liquid, and measures mycelia polysaccharide yield；
(6) gained clear liquid will be centrifugated twice to merge, freeze concentration, citric acid extraction displacement heavy metal ion, alcohol precipitation
Alcohol washes, is lyophilized and obtains qualified Fuscoporia obliqua polysaccharide product；
(7) gained Inonotus obliquus mycelia polysaccharide and exocellular polysaccharide are all made of Phenol sulfuric acid procedure measurement, and gained Inonotus obliquus is more
Sugar is the sum of mycelia polysaccharide and exocellular polysaccharide, is total starches.
(8) it referring to national standard, measures the pollutant lead in Fuscoporia obliqua polysaccharide, arsenic, mercury, cadmium, chromium, aflatoxin B1, take off
Oxygen nivalenol, carbendazim, avermectin, the content of butachlor.
In one aspect of the invention, Inonotus obliquus new strains JSU LIUK18 used in step (3) of the present invention is
The one plant of Inonotus obliquus new strains obtained through ultraviolet mutagenesis, can in wheat bran rice bran complete feed liquid culture medium fast-growth and height
The Inonotus obliquus new strains for producing fermentation material, are determined as one plant of birch through Sangon Biotech's DNA sequencing
Brown pore fungi new strains, the new strains are deposited in the Chinese Typical Representative positioned at Wuhan, China Wuhan University on December 19th, 2018
Culture collection (CCTCC), deposit number are CCTCC NO:M 2018906, and the Latin literary fame of recommendation is known as Inonotus obliquus
JSU LIUK18 Inonotus obliquus JSU LIUK18, the subsequent new bacterium of Inonotus obliquus of the invention patent specification
Strain refers both to this bacterial strain；
In one aspect of the invention, the charge in step (3) of the present invention when shaking flask is the 40% of shaking flask volume, is connect
Kind amount is the 10% of stocking volume, revolving speed 150r/min, incubation time 5-12d.
In one aspect of the invention, sample-loading amount is fermenter volume when upper tank fermentation in step (3) of the present invention
75%, ventilation quantity is that the ratio between ventilation volume and tinning liquid volume per minute are 1:0.8, cultivation temperature under aeration condition when upper tank
24-28 DEG C, stirring rate 50-150r/min, upper tank fermentation 5-12d.
Beneficial effects of the present invention
Using the Inonotus obliquus new strains, in the liquid for meeting the rice bran wheat bran complete feed of contaminants in foods through examining
Fermenting and producing Fuscoporia obliqua polysaccharide in culture medium, when the total concentration of rice bran and wheat bran reaches 10.1g/ in rice bran wheat bran complete feed culture medium
100mL, KH2PO4For 0.25g/100mL, MgSO4·7H2It is dry mycelial weight, mycelia polysaccharide, extracellular more when O is 0.25g/100mL
4.106g/100mL, 343.456mg/100mL and 663.514mg/100mL, total starches are respectively reached when sugared concentration shake flask fermentation
For 1006.970mg/100mL；And then can reach when the fermentation of upper tank 3.916g/100mL, 323.355mg/100mL,
641.724mg/100mL the level with 965.079mg/100mL.Through detecting, total starches that shake flask fermentation and upper tank ferment
Mycotoxin aflatoxin B1, the limitation of B2 and deoxynivalenol, the limit of heavy metal lead, arsenic, mercury, cadmium, chromium
Amount and pesticide carbendazim, avermectin, butachlor residue limits be not higher than GB2761-2017, GB2762-2017,
Limited Doses as defined in GB2763-2016 are not detected.
The method that the present invention realizes the Inonotus obliquus new strains production Fuscoporia obliqua polysaccharide for the first time, birch is brown in the method
Pore fungi new strains ferment in the liquid culture medium through examining the rice bran wheat bran complete feed for meeting contaminants in foods, ferment
Product removes the miscellaneous birch for obtaining qualification through multiple frequency ultrasonic extraction, freeze concentration, citric acid extraction displacement heavy metal ion, alcohol precipitation alcohol
Obliquus polysaccharide product.Used Inonotus obliquus new strains are one plant of new bacterium of Inonotus obliquus that inventor oneself mutagenesis obtains
Strain, the bacterial strain can effectively be grown in the new culture medium of liquid rice bran wheat bran complete feed, and Efficient Conversion rice bran wheat bran is that Inonotus obliquus is more
Sugar, the bacterial strain is unique, thus the present invention is also unique.
The present invention also achieves the novelty of substantive distinguishing features and marked improvement.Inonotus obliquus new strains Inonotus
The total concentration of rice bran and wheat bran reaches in the rice bran wheat bran complete feed liquid culture medium of obliquus CCTCC NO:M 2018906
When 10.1g/100mL, the Thick many candies concentration in the dry weight concentrations of mycelia, mycelia polysaccharide concentration and fermentation liquid is respectively reached
4.106g/100mL, 343.456mg/100mL and 663.514mg/100mL, total starches are that final Fuscoporia obliqua polysaccharide produces
Object reaches the production level of 1006.970mg/100mL, illustrates that Inonotus obliquus new strains fermenting property in Optimal Medium is good
And high yield Fuscoporia obliqua polysaccharide, the invention patent achieve the novelty with substantive distinguishing features and marked improvement.
The rice bran wheat bran of low value is converted to the Fuscoporia obliqua polysaccharide with plurality of health care functions of high level by the present invention, to drop
Low production cost, the reasonable higher value application of low side resource high-efficiency, protection health of masses, all with beneficial society and economic effect
Fruit, thus there is practicability.
In conclusion the present invention has had been provided with uniqueness necessary to authorizing patent of invention, has achieved substantive distinguishing features
With the innovation and practicality of marked improvement, technology, the beneficial effect of economy and society that patent of invention should have are produced.
Detailed description of the invention
Fig. 1 is the flow chart of the ultraviolet mutagenesis selection of present invention Inonotus obliquus new strains obtained.
Fig. 2 is the chadogram of present invention Inonotus obliquus new strains obtained.
Fig. 3 is that 2018906 shaking flask liquid state fermentation rice bran wheat bran complete feed of bacterial strain Inonotus obliquus CCTCC NO:M produces the brown hole of birch
The fermentation material of granulose.
Specific embodiment
The present invention attached process shown in FIG. 1 to specifications, provides through ultraviolet mutagenesis, breeding is in wheat bran rice bran complete feed
On liquid culture medium the speed of growth faster, the method for the higher Inonotus obliquus new strains of polysaccharide yield, the method includes following
Step:
The original strain Inonotus obliquus of laboratory preservation is taken, deposit number is that CFCC 83414 is to set out
Bacterial strain；
Taken starting strain is inoculated in PDA culture medium and carries out activation culture；
It is eluted after thallus culture with sterile saline and spore suspension is made；
Multiple needed for spore suspension is diluted after irradiation carries out mutagenesis in the UV lamp, is connected to the screening training of rice bran wheat bran
It supports and is protected from light culture on base, initial screening goes out the bacterial strain that growth rate is very fast, more stable；The bacterial strain of primary dcreening operation is subjected to genetic stability
Test, secondary screening select the bacterial strain that growth rate is very fast, more stable；
Shake flask fermentation is carried out with rice bran wheat bran complete feed liquid culture medium, growth rate height is filtered out and polysaccharide yield is high
Bacterial strain；
The new strains filtered out carry out antagonistic effect and gene sequencing.
In one embodiment, PDA culture medium used is potato 200g/L, glucose 20g/L, peptone 5g/
L, potassium dihydrogen phosphate 1.5g/L, epsom salt 0.75g/L, agar 20g/L, pH are natural.
In one embodiment, the constant temperature is 28 DEG C.
In one embodiment, the ultraviolet mutagenesis is secretly operated using feux rouges, in the ultraviolet lamp for being 20W apart from power
40s is irradiated under 20cm respectively.
In one embodiment, described to be protected from light culture to be protected from light culture 7 days at 28 DEG C.
In one embodiment, the culture used medium that is protected from light is rice bran, wheat bran solid plate culture medium: rice bran
20g/L, wheat bran 30g/L, potassium dihydrogen phosphate 1.5g/L, epsom salt 0.75g/L, agar 20g/L, pH are naturally, wheat bran, rice bran
Complete feed uses, and wheat bran rice bran has to comply with standard).
In one embodiment, the screening technique is plate diameter measuring method.
In one embodiment, the primary dcreening operation step are as follows: it is good to be protected from light picking growth on the plate of culture from ultraviolet mutagenesis
Good single colonie is seeded to respectively in new rice bran wheat bran solid plate culture medium, is therefrom selected and is grown relative to starting strain
Fastly, the high bacterial strain of shapeliness, stability.
In one embodiment, the secondary screening step are as follows: the more excellent bacterial strain that scalping is selected is subjected to 5 generation plates respectively and is passed
It is commissioned to train feeding, the well-grown single colonie of picking is seeded to respectively in new rice bran wheat bran solid plate culture medium, picks out growth
Speed is fast, healthy and strong, pure variant.
In one embodiment, the three sieves step are as follows: using the Seedling height rate variant that secondary screening determines as three sieves
Object, together with starting strain carry out shake flask fermentation screening so that it is determined that variant merit stablizes the bacterial strain of expression.It will
The Inonotus obliquus new strains and original starting strain Inonotus obliquus that secondary screening is selected, deposit number are CFCC 83414
Shake flask fermentation test is carried out, continuously fermented for 5 generations, determines target mutagenic fungi by index.
In one embodiment, the Inonotus obliquus mutagenesis new strains CCTCC NO:M 2018906 is No. 27 bacterium
Strain, the growth rate on the 5th generation plate of secondary screening are significantly faster than that the growth rate of No. 0 bacterial strain of starting strain.
In one embodiment, the shaking flask is 250mL conical flask.
In one embodiment, the rice bran, wheat bran liquid fermentation medium are as follows: rice bran 2.0g/100mL, wheat bran
3.0g/100mL, potassium dihydrogen phosphate 0.15g/100mL, magnesium sulfate 0.075g/100mL, pH are natural.
In one embodiment, the index is pure dry mycelial weight, mycelia polysaccharide weight, the hair of 100mL fermentation volume
Zymotic fluid crude extracellular polysaccharide weight and total starches weight.
In one embodiment, inhereditary feature is characterized with antimicrobial experiment, result is as shown in Fig. 2, and antagonism line is bright
Aobvious, the Inonotus obliquus mutagenesis new strains CCTCC NO:M 2018906 on the right grows dense, thick and solid, it is seen that its growth performance is better than
Beneficial variation has occurred in the performance of starting strain, inhereditary feature.
In one embodiment, Inonotus obliquus new strains, deposit number are CCTCC NO:M 2018906, raw through raw work
Object engineering (Shanghai) limited liability company DNA sequencing is determined as one plant of Inonotus obliquus new strains.
The new strains are deposited in the Chinese Typical Representative culture positioned at Wuhan, China Wuhan University on December 19th, 2018
Object collection (CCTCC), deposit number are CCTCC NO:M 2018906, and the Latin literary fame of recommendation is known as Inonotus obliquus JSU
LIUK18 Inonotus obliquus JSU LIUK18, the subsequent Inonotus obliquus new strains of the invention patent specification are equal
Refer to this bacterial strain (see Fig. 2).
The present invention provides a kind of method using mutagenic obtained Inonotus obliquus new strains production Fuscoporia obliqua polysaccharide, In
Inonotus obliquus new strains are through examining in the liquid culture medium of rice bran wheat bran complete feed for meeting contaminants in foods in this method
It ferments, after the fermentation material for producing Inonotus obliquus mycelium raw material and fermentation liquid, by multiple frequency ultrasonic assisted extraction, freezing
Concentration, citric acid extraction displacement heavy metal ion, alcohol precipitation alcohol wash, be lyophilized and obtains the Fuscoporia obliqua polysaccharide product of qualification, the side
Method includes the following steps:
In one embodiment, examine wheat bran and rice bran, Heavy Metals lead, arsenic, mercury, cadmium, chromium level of pollution most
Height is no more than 3mg/kg, remaining corresponds with the former Ministry of Agriculture and eats feed wheat bran standard NY/T 3218-2018 and rice bran standard
NY/T122-1989, applying pesticides meet agrotechnical specification during wheat planting, without serious head blight evil in wheat growth,
Harvesting season causes wheat rotten without continuous rainwater, and wheat bran storage and transportation is applied during not will lead to mildew Rice Cropping after wheat processing
Meet agrotechnical specification with pesticide, paddy growth is in the process without serious plant disease, and harvesting season causes paddy rotten without continuous rainwater, rice
Rice bran storage and transportation not will lead to mildew after paddy processing；
In one embodiment, the rice bran, wheat bran liquid fermentation medium are as follows: rice bran concentration is 0.1-10g/
100mL, wheat bran concentration are 0.1-10g/100mL, and the total concentration of rice bran and wheat bran is up to 10.1g/100mL, adds di(2-ethylhexyl)phosphate
Hydrogen potassium 0.015-0.25g/100mL, epsom salt 0.015-0.25g/100mL, the pH 100mL naturally, 250mL shaking flask charges,
Fermentor filling rate 75%, inoculum concentration 10%, 24-28 DEG C of cultivation temperature, shaking speed 150r/min or fermentation stirring slurry revolving speed
50-150r/min, fermentation time 5-12d；
In one embodiment, the resulting Inonotus obliquus mycelium of Liquid Culture is centrifuged, is respectively obtained
Mycelium and separating liquid measure crude extracellular polysaccharide to separation clear liquid Phenol sulfuric acid procedure；
In one embodiment, by Inonotus obliquus mycelium extract polysaccharide operation, with 60 DEG C of hot water add 35kHz and
The ultrasonic wave added liquid circulation of 1.5kW extracts 1.5h, and centrifuge separation takes clear liquid, and measures mycelia polysaccharide yield with phenol sulfuric acid；
In one embodiment, gained fermentation liquid and mycelia extracting solution merge, and set by freeze concentration, citric acid extraction
Change heavy metal ion, alcohol precipitation alcohol is washed, the method that is freeze-dried obtains qualified exocellular polysaccharide；
In one embodiment, the fermentation screened obtained Inonotus obliquus new strains and fermentation medium is utilized to obtain
As a result are as follows: the Thick many candies concentration in the dry weight concentrations of mycelia, mycelia polysaccharide concentration and fermentation liquid respectively reach 4.106g/100mL,
343.456mg/100mL and 663.514mg/100mL, Fuscoporia obliqua polysaccharide 1006.970mg/100mL.
In one embodiment, Fuscoporia obliqua polysaccharide yield reaches 1006.970mg/100mL, illustrates that Inonotus obliquus is new
Bacterial strain fermenting property in Optimal Medium is good and high yield Fuscoporia obliqua polysaccharide, the invention patent are achieved with substantive distinguishing features
With the novelty of marked improvement.
In one embodiment, through detecting, mycotoxin aflatoxin B1, B2 and the deoxynivalenol of total starches
Limitation, heavy metal lead, arsenic, mercury, cadmium, the limitation of chromium and the pesticide carbendazim of bacterium enol, avermectin, butachlor residue limits
It is not higher than Limited Doses as defined in GB2761-2017, GB2762-2017, GB2763-2016 or is not detected.
Embodiment one
Using Inonotus obliquus new strains.Wheat bran and rice bran is examined to meet use standard of the invention.The usage amount of wheat bran
For 10.0g/100mL, the usage amount of rice bran is that 0.1g/100mL adds KH2PO40.25g/100mL, MgSO4·7H2O
0.25g/100mL, pH are naturally, add water to required volume, and 121 DEG C of sterilizing 30min, as culture medium for fermenting, shaking flask is filled
Sample amount is 40%, inoculum concentration 10%, 28 DEG C of cultivation temperature, revolving speed 150r/min, incubation time 12d.Aforementioned invention is pressed below
Implement step (3)~(7) in technical solution described by content.As a result are as follows: dry mycelial weight 4.106g/100mL, mycelia polysaccharide are
343.456mg/100mL exocellular polysaccharide 663.514mg/100mL, Inonotus obliquus total starches are 1006.970mg/100mL.
Through detecting, the mycotoxin aflatoxin B1 of total starches, the limitation of B2 and deoxynivalenol, heavy metal lead, arsenic,
Mercury, cadmium, the limitation of chromium and pesticide carbendazim, avermectin, butachlor residue limits be not higher than GB2761-2017,
Limited Doses as defined in GB2762-2017, GB2763-2016 are not detected.
Embodiment two
Using Inonotus obliquus new strains.Wheat bran and rice bran is examined to meet use standard of the invention.Rice bran usage amount is
5.0g/100mL culture medium, the usage amount of wheat bran are 5.1g/100mL culture medium, add KH2PO40.15g/100mL,
MgSO4·7H2Naturally, adding water to required volume, 121 DEG C of sterilizing 30min are used for as culture medium by O 0.15g/100mL, pH
Fermentation, shaking flask sample-loading amount are 40%, inoculum concentration 10%, 24 DEG C of cultivation temperature, revolving speed 150r/min, incubation time 5d.Below
Implement by step (3)~(7) in technical solution described by foregoing summary.As a result are as follows: dry mycelial weight 2.878g/
100mL, mycelia polysaccharide 207.182mg/100mL, exocellular polysaccharide 964.165mg/100mL, Fuscoporia obliqua polysaccharide are
1171.347mg/100mL；Through detecting, the mycotoxin aflatoxin B1s of total starches, B2 and deoxynivalenol
Limitation, heavy metal lead, arsenic, mercury, cadmium, the limitation of chromium and pesticide carbendazim, avermectin, butachlor residue limits be not higher than
Limited Doses as defined in GB2761-2017, GB2762-2017, GB2763-2016 are not detected.
Embodiment three
Using Inonotus obliquus new strains.Wheat bran and rice bran is examined to meet use standard of the invention.Rice bran usage amount is
2.0g/100mL culture medium, the usage amount of wheat bran are 8.1g/100mL culture medium, add KH2PO40.015g/100mL,
MgSO4·7H2Naturally, adding water to required volume, 121 DEG C of sterilizing 30min are used for as culture medium by O 0.015g/100mL, pH
Fermentation, shaking flask sample-loading amount are 40%, inoculum concentration 10%, 27 DEG C of cultivation temperature, revolving speed 150r/min, incubation time 6d.Below
Implement by step (3)~(7) in technical solution described by foregoing summary.As a result are as follows: dry mycelial weight 2.521g/
100mL, mycelia polysaccharide 365.898mg/100mL, exocellular polysaccharide 1126.054mg/100mL, Fuscoporia obliqua polysaccharide are
1542.8mg/100mL；Through detecting, mycotoxin aflatoxin B1, the limit of B2 and deoxynivalenol of total starches
Amount, heavy metal lead, arsenic, mercury, cadmium, the limitation of chromium and pesticide carbendazim, avermectin, butachlor residue limits be not higher than
Limited Doses as defined in GB2761-2017, GB2762-2017, GB2763-2016 are not detected.
Example IV
Using Inonotus obliquus new strains.Wheat bran and rice bran is examined to meet use standard of the invention.Rice bran usage amount is
0.1g/100mL culture medium, the usage amount of wheat bran are 10.0g/100mL culture medium, add KH2PO40.2g/100mL,
MgSO4·7H2Naturally, adding water to required volume, 121 DEG C of sterilizing 30min are used for as culture medium by O 0.18g/100mL, pH
Fermentation, shaking flask sample-loading amount are 40%, inoculum concentration 10%, 27 DEG C of cultivation temperature, revolving speed 150r/min, incubation time 7d.Below
Implement by step (3)~(7) in technical solution described by foregoing summary.As a result are as follows: dry mycelial weight 2.553g/
100mL, mycelia polysaccharide 289.072mg/100mL, exocellular polysaccharide 1149.181mg/100mL, Fuscoporia obliqua polysaccharide are
1438.253mg/100mL；Through detecting, the mycotoxin aflatoxin B1s of total starches, B2 and deoxynivalenol
Limitation, heavy metal lead, arsenic, mercury, cadmium, the limitation of chromium and pesticide carbendazim, avermectin, butachlor residue limits be not higher than
Limited Doses as defined in GB2761-2017, GB2762-2017, GB2763-2016 are not detected.
Embodiment five
Using Inonotus obliquus new strains.Wheat bran and rice bran is examined to meet use standard of the invention.Rice bran usage amount is
3.5g/100mL culture medium, the usage amount of wheat bran are 6.6g/100mL culture medium, add KH2PO40.05g/100mL,
MgSO4·7H2Naturally, adding water to required volume, 121 DEG C of sterilizing 30min are used for as culture medium by O 0.05g/100mL, pH
Fermentation, shaking flask sample-loading amount are 40%, inoculum concentration 10%, 28 DEG C of cultivation temperature, revolving speed 150r/min, incubation time 6d.Below
Implement by step (3)~(7) in technical solution described by foregoing summary.As a result are as follows: dry mycelial weight 2.511g/100mL
Culture medium, mycelia polysaccharide are 371.688mg/100mL culture medium, and exocellular polysaccharide is 1062.454mg/100mL culture medium, and birch is brown
Obliqua polysaccharide is 1434.143mg/100mL culture medium；Through detecting, the mycotoxin aflatoxin B1s of total starches, B2 and de-
The limitation of oxygen nivalenol, heavy metal lead, arsenic, mercury, cadmium, the limitation of chromium and pesticide carbendazim, avermectin, butachlor
Residue limits be not higher than Limited Doses as defined in GB2761-2017, GB2762-2017, GB2763-2016 or be not detected.
Embodiment six
Using Inonotus obliquus new strains.Wheat bran and rice bran is examined to meet use standard of the invention.Fermentor sample-loading amount
It is the 75% of fermenter volume, fermentation temperature is 24 DEG C, ventilation quantity 1:0.8v/v/mim, mixing speed 50r/min, tank gauge pressure
0.05MPa, inoculum concentration 10%, incubation time 5d, fermentation medium are wheat bran 10.0g/100mL, and rice bran usage amount is 0.1g/
100mL culture medium, KH2PO40.25g/100mL, MgSO4·7H2O 0.25g/100mL, pH are natural.Below by aforementioned invention
Implement step (3)~(7) held in described technical solution.As a result are as follows: dry mycelial weight 3.892g/100mL, mycelia polysaccharide are
307.921mg/100mL, exocellular polysaccharide 617.835mg/100mL, Fuscoporia obliqua polysaccharide 925.756mg/100mL；Through examining
Survey, the mycotoxin aflatoxin B1 of total starches, the limitation of B2 and deoxynivalenol, heavy metal lead, arsenic, mercury,
Cadmium, the limitation of chromium and pesticide carbendazim, avermectin, butachlor residue limits be not higher than GB2761-2017, GB2762-
2017, it Limited Doses as defined in GB2763-2016 or is not detected.
Embodiment seven
Using Inonotus obliquus new strains.Wheat bran and rice bran is examined to meet use standard of the invention.Fermentor sample-loading amount
It is the 75% of fermenter volume, fermentation temperature is 24 DEG C, ventilation quantity 1:0.8v/v/mim, mixing speed 50r/min, tank gauge pressure
0.05MPa, inoculum concentration 10%, incubation time 12d, fermentation medium be rice bran 10.0g/100mL, wheat bran 0.1g/100mL,
KH2PO40.015g/100mL, MgSO4·7H2O 0.015g/100mL, pH are natural.Below by skill described by foregoing summary
Implement step (3)~(7) in art scheme.As a result are as follows: dry mycelial weight 2.418g/100mL, mycelia polysaccharide 277.13mg/
100mL, exocellular polysaccharide 1008.371mg/100mL, Fuscoporia obliqua polysaccharide 1285.501mg/100mL；Through detecting, total starches
Mycotoxin aflatoxin B1, the limitation of B2 and deoxynivalenol, the limit of heavy metal lead, arsenic, mercury, cadmium, chromium
Amount and pesticide carbendazim, avermectin, butachlor residue limits be not higher than GB2761-2017, GB2762-2017,
Limited Doses as defined in GB2763-2016 are not detected.
Embodiment eight
Using Inonotus obliquus new strains.Wheat bran and rice bran is examined to meet use standard of the invention.Fermentor sample-loading amount
It is the 75% of fermenter volume, fermentation temperature is 28 DEG C, ventilation quantity 1:0.8v/v/mim, mixing speed 150r/min, tank gauge pressure
0.05MPa, inoculum concentration 10%, incubation time 5d, fermentation medium are rice bran 2.5g/100mL, wheat bran 7.5g/100mL, KH2PO4
0.18g/100mL, MgSO4·7H2O 0.18g/100mL, pH are natural.Below by technical solution described by foregoing summary
The step of (3)~(7) implement.As a result are as follows: dry mycelial weight 2.396g/100mL, mycelia polysaccharide 391.478mg/100mL, it is extracellular
Polysaccharide is 1234.511mg/100mL, Fuscoporia obliqua polysaccharide 1625.989mg/100mL；Through detecting, the mycotoxin of total starches
Aflatoxin B1, the limitation of B2 and deoxynivalenol, heavy metal lead, arsenic, mercury, cadmium, the limitation of chromium and pesticide are more
Bacterium spirit, avermectin, butachlor residue limits be not higher than GB2761-2017, GB2762-2017, GB2763-2016 rule
Fixed Limited Doses are not detected.
Claims (1)
1. utilizing the method for mutagenic obtained Inonotus obliquus bacterial strain production Fuscoporia obliqua polysaccharide, it is characterised in that as steps described below
It carries out:
(1) wheat bran and rice bran are examined；
(2) rice bran and wheat bran are added into 121 DEG C of sterilizing 30min of the desired amount of water；
(3) fermentation medium of Inonotus obliquus new strains is directly used in using rice bran and wheat bran complete feed as carbon source, nitrogen source, rice bran is dense
Degree is 0.1-10g/100mL, and wheat bran concentration is 0.1-10g/100mL, and the total concentration of rice bran and wheat bran is up to 10.1g/
100mL, add potassium dihydrogen phosphate 0.015-0.25g/100mL, epsom salt 0.015-0.25g/100mL, pH naturally,
250mL shaking flask charging 100mL, fermentor filling rate 75%, inoculum concentration 10%, 24-28 DEG C of cultivation temperature, shaking speed 150r/
Min or fermentation rotating speed of agitator 50-150r/min, fermentation time 5-12d；
(4) liquid culture is centrifugally separating to obtain mycelium and fermentation liquid, and measures the yield of exocellular polysaccharide in fermentation liquid；
(5) the ultrasonic wave added liquid circulation of 35kHz and 1.5kW is added to extract 1.5h, centrifugation point 60 DEG C of hot water of gained mycelium
From taking clear liquid, and measure mycelia polysaccharide yield；
(6) gained clear liquid will be centrifugated twice to merge, freeze concentration, citric acid extract displacement heavy metal ion, alcohol precipitation alcohol is washed,
Freeze-drying obtains qualified Fuscoporia obliqua polysaccharide product.
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