CN110283861A - A method of ganoderma lucidum polysaccharide is produced using the ganoderma strain that mutagenesis generates - Google Patents
A method of ganoderma lucidum polysaccharide is produced using the ganoderma strain that mutagenesis generates Download PDFInfo
- Publication number
- CN110283861A CN110283861A CN201910608915.3A CN201910608915A CN110283861A CN 110283861 A CN110283861 A CN 110283861A CN 201910608915 A CN201910608915 A CN 201910608915A CN 110283861 A CN110283861 A CN 110283861A
- Authority
- CN
- China
- Prior art keywords
- ganoderma lucidum
- rice bran
- fermentation
- wheat bran
- polysaccharide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/01—Preparation of mutants without inserting foreign genetic material therein; Screening processes therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
Landscapes
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Plant Pathology (AREA)
Abstract
The invention discloses a kind of methods of ganoderma strain production ganoderma lucidum polysaccharide generated using mutagenesis, are related to food microorganisms applied technical field.Ganoderma lucidum 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 ganoderma lucidum 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 ganoderma lucidum polysaccharide product of qualification.Present invention employs a kind of new methods; by the rice bran wheat bran of low value by ganoderma lucidum liquid deep fermentation be converted to can food ganoderma lucidum polysaccharide; reduction production cost, low side resource high-efficiency are rationally utilized, protect health of masses etc.; all there is beneficial society and economic effect, thus unique, innovation and practicality.
Description
Technical field
The present invention relates to food microorganisms applied technical field more particularly to a kind of use mutagenic obtained ganoderma lucidum new strains
The method for producing ganoderma lucidum polysaccharide, ganoderma lucidum new strains are through examining the rice bran wheat bran for meeting contaminants in foods complete in the method
It ferments in the liquid culture medium of material, after the fermentation material for producing ganoderma lucidum mycelium raw material and fermentation liquid, by multiple frequency ultrasonic
Assisted extraction, freeze concentration, citric acid extraction displacement heavy metal ion, alcohol precipitation alcohol wash, be lyophilized and obtains qualified ganoderma lucidum polysaccharide production
Object.
Background technique
Dual-purpose of drug and food fungi has long usage history to use crowd with extensive in China.Modern science discloses, existing
The energy in common dual-purpose of drug and food fungi such as ganoderma lucidum, Hericium erinaceus, cordyceps sinensis, the mycelium of grifola frondosus, fructification or spore
Generate a variety of active ingredients such as amino acid, protein, vitamin, polysaccharide, ucleosides, flavonoids or enrichment lot of trace member
Plain such as selenium, zinc have and improve the immunity of the human body, is antitumor, enhancing the multiple efficacies such as liver function, anti-oxidant.Just because of such as
This, the enthusiasm of international and domestic research and development dual-purpose of drug and food fungi is in the ascendant, in field of medicaments, field of food, Traditional Chinese health
All there are many researcher and developers for field, agricultural and field of industrial production, obtain numerous research achievement or product.With regard to rare
Edible mushroom is used for for food, and domestic market has edible mushroom health-care food such as ganoderma lucidum capsule, grifola frondosus capsule and ganoderma lucidum capsule
Deng in fast sale, their economic value is very high.Hericium erinaceus is added in biscuit, Hericium erinaceus biscuit has then been made, have become common food
Product at home sold by supermarket.New Zealand, Japan, the U.S., South Korea etc., which produce, in the world similar edible mushroom health-care food
Product.
For ganoderma lucidum, research shows that: ganoderma lucidum, which has, such as protects liver, treatment diabetes, treatment cardiovascular, anti-swollen
Tumor, enhancing are immune and other effects, and (bibliography: 1. Li Ying wave is efficiently separated from ganoderma lucidum mycelium prepares antitumor ganoderic acid list
The Shanghai research [D] of body: East China University of Science, 2013. 2. Yuen J W M, Mak D S Y, Chan E S, et
al.Tumor inhibitory effects of intravesical Ganoderma lucidum instillation in
the syngeneic orthotopic MB49/C57bladder cancer mice model[J].Journal of
Ethnopharmacology,2018:S0378874118311991.③Yongjun Kan,Tiqiang Chen,Yanbin
Wu,Jianguo wu,Jinzhong Wu,et al.Antioxidant activity of polysaccharide
extracted from Ganoderma lucidum using response surface methodology[J]
.International the 4. precious pine of Teng of Journal of Biological Macromolecules 72 (2015) 151-157.
The screening of the effective hypoglycemic component of ganoderma lucidum and the Shanghai hypoglycemic mechanism study [D]: Fudan University, 2011. 5. Tang W, Liu J
W,Zhao W M,et al.Ganodermic acid T from Ganoderma lucidum mycelia induces
mitochondria mediated apoptosis in lung cancer cells[J].Life Sciences,2006,80
(3):0-211.)。
Ganoderma lucidum (Ganoderma lucidum) belongs to Basidiomycota, Basidiomycetes, agaric subclass, Aphyllophorales, Ganoderma Lucidum
Section, Ganoderma are a kind of dual-purpose of drug and food macro fungis, be born in woods by the timber of broad leaf tree more or wood, standing tree, fall wood on,
Sometimes it is also born in coniferous tree, cap suberin, kidney shape or semicircle, reddish brown, purple or mulberry, tool paint sample gloss has ring
Shape rib and radial wrinkle, size and metamorphosis are very big, and the cap of Large scale individual is 20 × 10cm, thickness about 2cm, general a
Body is 4 × 3cm, and 0.5~1cm of thickness has numerous small holes below, and nozzle is white or filbert, has 4~5 in every millimeter, nozzle
Circle, inner wall are hymenium, and spore results from load top.Stem side is raw, seldom wilfully, is longer than bacteria cover diameter, puce is extremely
Black has paint sample gloss, hard.Spore oval, 8~11 × 7cm, two layers of wall, there is small wart on inner wall brown, surface, and outer wall is saturating
It is bright colourless.Ganoderma lucidum Europe, America, Africa, east Asia etc. area be distributed, China ganoderma lucidum be mainly distributed Longquan, Zhejiang Province,
There is part on the ground such as Heilungkiang, Jilin, Hebei, Shandong, Huoshan, Jiangsu, Jiangxi, Hunan, Guizhou, Fujian, Guangdong, Guangxi
Yield.The ganoderma lucidum speed of growth under wild state is very slow, and price is high.As it can be seen that since Wild ganoderma resource is very rare, now
The acquisition modes of ganoderma lucidum are mainly artificial cultivation, but the artificial cultivation period is long, production efficiency is not high, large labor intensity, by season,
The limitation such as environment, is subject to pest and disease damage, quality is unstable with yield.Rare fungi fermentation technology, can overcome traditional fructification to plant
The deficiency of training, using liquid or the method for solid state fermentation production Ganoderma fermented product and be subject to deep processing application meet innovation driving
Thinking of development has good prospect.The progress obtained to the research and development of Ganoderma fermented product all will further push ganoderma lucidum
Application, bring actual social and economic benefit.Just because of this, just add it is necessary to study new Ganoderma fermented product and its deeply
The production method of chemical product, for example, consider by some low side processing of farm products byproduct sources such as wheat brans, rice bran Efficient Conversion be
The glossy ganoderma fermentation raw material of high value, it is expected to reduce the production cost of Ganoderma fermented product to reduce finished product such as ganoderma lucidum 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.Rice bran and wheat bran are as paddy or small
The by-product of Meccah work, rich in nutrition content are cheap.Contain in rice bran and enriches good protein, active polysaccharide, fat
Active material significant with the physiological functions such as tocotrienols, tocopherol, wheat bran (referring to wheat bran, full text is herewith) are rich in rouge
The nutritional ingredients such as fat, protein, minerals, vitamin and cellulose, wherein the content of cellulose up to wheat bran total amount 18%
More than, so, rice bran and wheat bran can grow for ganoderma lucidum and provide sufficient carbon source, nitrogen source, vitamin and mineral.In ganoderma fiber
Under the action of plain enzyme and other enzyme systems, rice bran and wheat bran can be converted to needed nutrient matter and carry out growth metabolism, synthesis by ganoderma lucidum
Ganoderma lucidum mycelium and adenosine class functional materials.Therefore, the carbon source and nitrogen source with agricultural and sideline product rice bran, wheat bran as whole,
It carries out Ganoderma lucidum submerged fermentation and grows ganoderma lucidum class healthy food material, there is technical feasibility, and preferable society and warp can be brought
Ji benefit.
Most powder of edible fungus Liquid-state fermentation production methods use glucose, grain class raw material such as starch, potato or soya bean
Powder etc. is used as culture medium, even if using agricultural and sideline product such as rice bran, wheat bran etc., and is added as auxiliary element.Original food
With bacterial strains such as bacterium cordyceps sinensis, ganoderma lucidum, grifola frondosus in the rice bran wheat bran complete feed liquid for not adding glucose or other grain class raw materials
Upgrowth situation is not best on state culture medium, in addition rice bran, in bran feedstock may containing some harmful substances for example lead, arsenic,
Aflatoxin etc. may be transferred in mycelium, to prevent this transfer, therefore need the ferment rice bran wheat bran to such bacterium
Liquid complete feed method carries out innovation research.
The liquid state fermentation to grifola frondosus, ganoderma lucidum, cordyceps sinensis during the last ten years of the research group of inventor herein Liu Weimin
A large amount of research has been carried out, multiple achievements such as Master's thesis and patent of invention are successively formed.The Master's thesis of Liu Weimin guidance has:
(1) Yang Suohua, frondosa fermentation rice bran prepare polysaccharide (2006)；(2) Gu Huimin, grifola frondosus liquid in rice bran culture medium are trained
Support the research (2009) for producing polysaccharide and being enriched with organic selenium；(3) it opens and builds, physical method mutagenesis grifola frondosus liquid fermentation rice bran wheat bran produces
The research (2010) of polysaccharide；(4) Guo Chunmei, induction mutation of bacterium, the liquid fermentation rice bran wheat bran of grifola frondosus produce polysaccharide and selenium-rich is ground
Study carefully (2011)；(5) Li Yanan, grifola frondosus induction mutation of bacterium and fermentation and performance study (2013)；(6) Liu Lili, high level turn
Change the Ganoderma lucidum submerged fermentation and induction mutation of bacterium (2012) of rice bran wheat bran；(7) Guo Tianlong, lucidum strain mutagenesis and liquid state fermentation are high
Value converts full rice bran wheat bran research (2013)；(8) Chen Jing, cordyceps sinensis liquid fermentation higher value application rice bran wheat bran produce more
The research (2014) of sugar；(9) Zhang Xiaofei, ganoderma strain (Ganoderma lucidum CFCC6043) full rice bran of liquid fermentation
The research (2014) of wheat bran and active polysaccharide, etc..Liu Weimin has as the patent of invention of first invention people: (1) using rice bran
The method of wheat bran compound material and grifola frondosus mutagenic strain production polysaccharide, 20101010579048.4；(2) rice bran and wheat bran are used for
The Grifola frondosa strain of compound material production polysaccharide, 201010579078.5；(3) for ferment rice bran and wheat bran extracting solution production ash
The bacterial strain of tree flower polysaccharide, 201110150888.3；(4) bacterium for rice bran and wheat bran complete feed liquid state fermentation production grifolan
Strain, 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, the production method of (9) 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 is efficiently high-valued all around trans-utilization rice bran and wheat bran
Production this special topic of Rare edible fungus fermentation material is studied, and realizes the imagination of innovation and creation, and is continuously improved various rare true
The production method of bacterium fermentation material.
The paper and invention relevant to the invention patent in the past that the inventor Liu Wei people comb instructed Master degree candidate
Patent consults institute's unanswerable problem when related ends are transferred the possession of in conjunction with enterprise, such as can not provide bacterial strain qualitative gene map
Critical support material is composed, the hope that enterprise carries out industrialization development is not strong etc., has been found that Industrialization to be moved towards, related
Patented technology and research paper there are still many problems needs research and solve and innovated.These problems include having difference
The acquisition and confirmation and exploitation of the rare fungus in high efficiency new strains of performance, the formulation of rice bran bran feedstock standard, rare fungi hair
The innovation of ferment object fermentation process, rare fungal fermentate isolate and purify new method use and these need to innovate the combination side of object
Formula etc..Patent of invention to uniqueness necessary to Grant Patent Right for Invention, acquirement substantive distinguishing features outstanding and is shown in design
The innovation and practicality for writing progress will give abundant consideration.So the combined innovation of above-mentioned these problems should be ensured that phase
The patent of invention of pass reaches uniqueness, innovation and practicality necessary to Grant Patent Right for Invention.
Inventor based on the above considerations and the research to Ganoderma fermented product production method, proposes the invention patent
Application is utilized the different ganoderma lucidum original strain of event fermenting property in ganoderma strain source different from the past, carries out mutagenic obtained new
Bacterial strain, and the ganoderma lucidum polysaccharide product that the production of ferment rice bran wheat bran is qualified.
In conclusion field of the present invention is that be used to ferment by ganoderma lucidum new strains wheat bran and rice bran complete feed produces ganoderma lucidum
Fermentation material simultaneously becomes qualified products after deep processing is handled.Be different from the achievement of previous research and acquirement: the present invention will solve
One of critical issue certainly is the method needed through mutagenesis, promotes the fermentability of original strain, obtains one kind and has not been reported
Ganoderma lucidum new strains, and can to this new strains carry out gene sequencing, it is ensured that strain used is correct；The invention solves key ask
The two of topic are to make wheat bran raw rice bran standard because wheat bran and rice bran source are complicated, wherein pollutant kind and its it is horizontal not
Together, inventor's previous studies and patent do not give enough concerns in this regard, bring indefinite consequence to subsequent product exploitation；
The invention solves critical issue three become qualified products after deep processing is handled for fermentation material mycelia and fermentation liquid
Production method.Above three promise the invention patent and other inventors and the present inventor Liu Weimin research team it is all kinds of
Achievement is 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 strain, obtains one kind and does not appear in the newspapers
The ganoderma lucidum new strains in road, and gene sequencing can be carried out to this new strains, it is ensured that strain used is correct.The background of this problem is,
Inventor team it is expected to obtain one plant from unused and with good performance ganoderma strain, and it is expected that it can be in wheat bran rice bran
There is good behaviour in terms of bioconversion, therefore considers the method for research glossy ganoderma fermentation wheat bran rice bran production Ganoderma fermented product.According to hair
The previous research experience of bright people, this kind of rare fungi fermentation wheat bran rice bran is firstly the need of the adaptability problem for solving strain.Therefore,
We need the fermentability to the lucidum strain in this production method to investigate, and are improved by the method for induction mutation of bacterium
Its fermenting property.In addition in the course of the research, to guarantee that mistake does not occur in strain after a large amount of strain processing step, it should into
Row gene sequencing.This does not attach great importance to this thing in previous research, meets again when to previous bacterial strain, row measures gene order again
Problem is arrived.In this way, this patent dig-ins the measurement of ganoderma lucidum gene series from the beginning, guarantee that bacterial strain is correct.
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
Enclose interior some exceptional values, such as certain determining heavy metals result excessive problem that previous studies do not measure, trace it to its cause be because
Complicated for wheat bran and raw rice bran source, quality is difficult to keep similar level, and unknown pollutant eventually enters into end product.In this way
Originally the invention production method designed for certain specific pollutants and its possible content, it is possible to occur it in the product
Its pollutant copes with the new pollutant being likely to occur so to redesign the production method of fermentation material.In this way, wheat bran and
The control of raw rice bran quality and standard formulation problem just become the problem of new invention Patent design must be taken into consideration.The present invention proposes
The use standard of raw material wheat bran and rice bran.
It is proposed the invention solves critical issue three for fermentation material carry out deep processing processing become qualification
The production method of product, background are that the conjunction of entire fermentation material production method is considered on the basis of the first two critical issue solves
Manage reasonability.Since wheat bran raw rice bran is possible to bring unknown pollutant into, in the medium in addition to being contemplated that addition pollutant
Complexing agent or the additive for facilitating degradation of contaminant can also after fermentation wash tunning citric acid complex, alcohol precipitation alcohol
Process for separation and purification obtains qualified ganoderma lucidum crude polysaccharide 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 ganoderma lucidum new strains by the method for mutagenesis.The selection of microorganism mainly has physics to lure
Change, chemical mutagenesis, genetic recombination etc., the present invention be avoid using poisonous and hazardous chemical mutagen and field of food also not by
The gene engineering method that masses approve will utilize relatively easy easy ultraviolet mutagenesis technology, starting strain made to be in mutagenesis
A possibility that extreme environment expands the range in mutational site to greatest extent, and raising obtains direct mutation bacterial strain.The present invention is using ultraviolet
This common method of mutagenesis of mutagenesis, although the method does not have novelty, the bacterial strain performance that the method obtains is created
The variation of new property, 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 ganoderma lucidum new strains in the liquid for meeting the rice bran wheat bran complete feed of contaminants in foods through examining
It ferments in state culture medium, the method for preparing ganoderma lucidum polysaccharide, concrete composition, fermentation condition including culture medium are extracted and gone
The measured value of miscellaneous method, mycelia and polysaccharide yield and the polysaccharide index of quality.The mycotoxin of ganoderma lucidum total starches obtained is yellow bent
The limitation of mould toxin B1, B2 and vomitoxin, heavy metal lead, arsenic, mercury, cadmium, the limitation of chromium and pesticide carbendazim, avermectin,
The residue limits of butachlor are limited the quantity not higher than as defined in GB2761-2017, GB2762-2017, GB2763-2016 by measurement
It is worth or is not detected.
Summary of the invention
A kind of method for producing ganoderma lucidum polysaccharide by mutagenic obtained ganoderma lucidum new strains of the present invention, in the method ganoderma lucidum
New strains ferment in the liquid culture medium through examining the rice bran wheat bran complete feed for meeting contaminants in foods, produce spirit
After the fermentation material of camphorata mycelium raw material and fermentation liquid, by multiple frequency ultrasonic assisted extraction, freeze concentration, citric acid extraction displacement weight
Metal ion, alcohol precipitation alcohol washes, is lyophilized and obtains qualified ganoderma lucidum polysaccharide product.
The technical solution used in the present invention is as follows: using a kind of ganoderma lucidum new strains obtained through ultraviolet mutagenesis, through examining
It tests in the liquid culture medium for meeting the rice bran wheat bran complete feed of contaminants in foods and ferments, produce ganoderma lucidum mycelium raw material
After the fermentation material of fermentation liquid, by multiple frequency ultrasonic assisted extraction, freeze concentration, citric acid extraction displacement heavy metal ion, alcohol
Heavy alcohol washes, is lyophilized and obtains qualified ganoderma lucidum polysaccharide product.
In one aspect of the invention, the method for the ganoderma lucidum new strains production ganoderma lucidum polysaccharide generated using mutagenesis, according to
Following step carries 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) rice bran and wheat bran complete feed ganoderma strain Ganoderma lucidum is directly used in as carbon source, nitrogen source to save
The fermentation medium of number CCTCC NO:M 2018908, rice bran concentration are 0.1-10g/100mL, and wheat bran concentration is 0.1-10g/
The total concentration of 100mL, rice bran and wheat bran is 10.1g/100mL, adds potassium dihydrogen phosphate 0.015-0.25g/100mL, seven water sulphur
Sour magnesium 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 rotating speed of agitator 50-150r/min, fermentation time 5-12d；
(3) liquid culture is centrifugally separating to obtain mycelium and fermentation liquid, and measures the production of exocellular polysaccharide in fermentation liquid
Amount；
(4) the ultrasonic wave added liquid circulation of 35kHz 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；
(5) 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 ganoderma lucidum polysaccharide product；
(6) gained Ganoderma lucidum mycelium polysaccharide and exocellular polysaccharide are all made of Phenol sulfuric acid procedure measurement, and gained ganoderma lucidum polysaccharide is mycelia
The sum of polysaccharide and exocellular polysaccharide are total starches.
(7) referring to national standard, pollutant lead, arsenic, mercury, cadmium, chromium, aflatoxin B1, the deoxidation snow in ganoderma lucidum polysaccharide are measured
Rotten sickle-like bacteria enol, carbendazim, avermectin, the content of butachlor.
In one aspect of the invention, ganoderma lucidum new strains used in step (3) of the present invention are inventors by laboratory
The ganoderma strain of preservation through Sangon Biotech's DNA sequencing through determining obtained by Uv-induced screening
For one plant of ganoderma lucidum new strains, the Chinese Typical Representative culture positioned at Wuhan, China Wuhan University is deposited on December 19th, 2018
Object collection (CCTCC), bacterial strain deposit number be CCTCC NO:M 2018908, it is proposed that classification naming be ganoderma lucidum JSU
LIUYU18:Ganoderma lucidum JSU LIUYU18, the subsequent ganoderma lucidum new strains of this specification refer 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 ganoderma lucidum new strains, in the liquid culture for meeting the rice bran wheat bran complete feed of contaminants in foods through examining
Fermenting and producing ganoderma lucidum polysaccharide in base, when in rice bran wheat bran complete feed culture medium the total concentration of rice bran and wheat bran reach 10.1g/100mL,
KH2PO4For 0.25g/100mL, MgSO4·7H2When O is 0.25g/100mL, dry mycelial weight, mycelia polysaccharide, exocellular polysaccharide concentration
5.722g/100mL, 669.861mg/100mL and 1402.065mg/100mL are respectively reached when shake flask fermentation, total starches are
2071.926mg/100mL；And it then can reach 5.384g/100mL, 602.3mg/100mL, 1327.8mg/ when the fermentation of upper tank
The level of 100mL and 1930.1mg/100mL.Through detecting, the mycotoxin for the total starches that shake flask fermentation and upper tank ferment is yellow
The limitation of aspertoxin B1, B2 and deoxynivalenol, heavy metal lead, arsenic, mercury, cadmium, the limitation of chromium and the more bacterium of pesticide
Spirit, avermectin, butachlor residue limits be not higher than as defined in GB2761-2017, GB2762-2017, GB2763-2016
Limited Doses are not detected.
The method that the present invention realizes the ganoderma lucidum new strains production ganoderma lucidum polysaccharide for the first time, ganoderma lucidum new strains exist in the method
It ferments in liquid culture medium through examining the rice bran wheat bran complete feed for meeting contaminants in foods, tunning is super through multifrequency
Sound extraction, freeze concentration, citric acid extraction displacement heavy metal ion, alcohol precipitation alcohol remove the miscellaneous ganoderma lucidum polysaccharide product for obtaining qualification.
Used ganoderma lucidum new strains are one plant of ganoderma lucidum new strains that inventor oneself mutagenesis obtains, which can be in liquid rice bran wheat bran
The new culture medium of complete feed is effectively grown, and Efficient Conversion rice bran wheat bran is ganoderma lucidum polysaccharide, and the bacterial strain is unique, thus the present invention
Also unique.
The present invention also achieves the novelty of substantive distinguishing features and marked improvement.The rice bran wheat bran complete feed liquid of ganoderma lucidum new strains
When the total concentration of rice bran and wheat bran reaches 10.1g/100mL in state culture medium, dry weight concentrations, mycelia polysaccharide concentration and the fermentation of mycelia
Thick many candies concentration in liquid respectively reaches 5.722g/100mL, 669.861mg/100mL and 1402.065mg/100mL, total starches
For 2071.926mg/100mL.Total starches are final ganoderma lucidum polysaccharide product, reach the production level of 2071.926mg/100mL,
Illustrate that ganoderma lucidum new strains fermenting property in Optimal Medium is good and high yield ganoderma lucidum polysaccharide, the invention patent are achieved with essence
The novelty of property feature and marked improvement.
The rice bran wheat bran of low value is converted to the ganoderma lucidum polysaccharide with plurality of health care functions of high level by the present invention, is given birth to reducing
Cost, the reasonable higher value application of low side resource high-efficiency, protection health of masses are produced, all there is beneficial society and economic effect, because
And 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
The shaking flask liquid state fermentation rice bran wheat bran complete feed that Fig. 1 is ganoderma lucidum new strains deposit number CCTCC NO:M 2018908 is trained
Support outside drawing when base.
Fig. 2 is ganoderma lucidum new strains deposit number CCTCC NO:M 2018908 in liquid state fermentation rice bran wheat bran complete feed culture medium
Resulting mycelia product figure.
Fig. 3 is the chadogram of ganoderma lucidum new strains.
Specific embodiment
The present invention provides a kind of methods using mutagenic obtained ganoderma lucidum new strains production ganoderma lucidum polysaccharide, in the method
Ganoderma lucidum new strains ferment in the liquid culture medium through examining the rice bran wheat bran complete feed for meeting contaminants in foods, produce
Out after the fermentation material of ganoderma lucidum mycelium raw material and fermentation liquid, set by multiple frequency ultrasonic assisted extraction, freeze concentration, citric acid extraction
Change heavy metal ion, alcohol precipitation alcohol is washed, is lyophilized and is obtained qualified ganoderma lucidum polysaccharide product, the 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；
The present invention provides by ultraviolet mutagenesis, breeding on wheat bran rice bran complete feed liquid culture medium the speed of growth faster,
The method of the higher ganoderma lucidum new strains of polysaccharide yield, the method includes the following steps:
Taking the original strain Ganoderma lucidum JSU LIU17 of laboratory preservation is starting 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 on base
It is protected from light culture, 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 inheritance stability
Property examination
It tests, secondary screening selects 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 30W apart from power
30s 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 using 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 ganoderma lucidum new strains and original starting strain that secondary screening is selected carry out shake flask fermentation test, continuously ferment for 5 generations, determine target by index
Mutagenic fungi.
In one embodiment, the ganoderma lucidum mutagenesis new strains Ganoderma lucidum is No. 2 bacterial strain, multiple
Sieving the growth rate on the 5th generation plate is 1.18cm/d, hence it is evident that is faster than 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 and the hair of 100mL fermentation volume
Zymotic fluid exocellular polysaccharide 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 ganoderma lucidum mutagenesis new strains growth on the right is dense, thick and solid, it is seen that its growth performance is better than the performance of starting strain, heredity
Beneficial variation has occurred in shape.
In one embodiment, ganoderma lucidum new strains, deposit number CCTCC NO:M 2018908, Jing Shenggong bioengineering
(Shanghai) limited liability company DNA sequencing is determined as one plant of ganoderma lucidum new strains.
The ganoderma lucidum new strains are deposited in the Chinese Typical Representative positioned at Wuhan, China Wuhan University on December 19th, 2018
Culture collection (CCTCC), deposit number be CCTCC NO:M 2018908, it is proposed that classification naming be ganoderma strain
JSU LIUYU18Ganoderma lucidum JSU LIUYU18, ganoderma lucidum new strains of the present invention refer both to this bacterial strain (see figure
3)。
In one embodiment, ganoderma lucidum new strains deposit number CCTCC NO:M 2018908 is through raw work bioengineering
(Shanghai) limited liability company DNA sequencing is determined as one plant of ganoderma lucidum new strains, is used as fermentation strain；
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 10.1g/100mL, add potassium dihydrogen phosphate
0.015-0.25g/100mL, epsom salt 0.015-0.25g/100mL, pH 100mL, fermentation naturally, 250mL shaking flask charges
Tank 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 ganoderma lucidum mycelium of Liquid Culture is centrifuged, respectively obtains mycelia
Body and separating liquid measure crude extracellular polysaccharide to separation clear liquid Phenol sulfuric acid procedure；
In one embodiment, by ganoderma lucidum 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 results for screening obtained ganoderma lucidum new strains and fermentation medium being utilized to obtain
Are as follows: the Thick many candies concentration in the dry weight concentrations of mycelia, mycelia polysaccharide concentration and fermentation liquid respectively reach 5.722g/100mL,
669.861mg/100mL and 1402.065mg/100mL, ganoderma lucidum polysaccharide 2071.926mg/100mL.
In one embodiment, ganoderma lucidum polysaccharide yield reaches 2071.926mg/100mL, illustrates ganoderma lucidum new strains excellent
Change the good simultaneously high yield ganoderma lucidum polysaccharide of fermenting property in culture medium, the invention patent is achieved with substantive distinguishing features and marked improvement
It is innovative.
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 ganoderma lucidum new strains.Wheat bran and rice bran is examined to meet use standard of the invention.The usage amount of rice bran is
0.1g/100mL culture medium, the usage amount of wheat bran are 10.0g/100mL culture medium, add KH2PO40.25g/100mL,
MgSO4·7H2Naturally, adding water to required volume, 121 DEG C of sterilizing 30min are used to send out as culture medium by O0.25g/100mL, pH
Ferment, shaking flask sample-loading amount are 40%, inoculum concentration 10%, 28 DEG C of cultivation temperature, revolving speed 150r/min, incubation time 5d.It presses below
Implement step (3)~(7) in technical solution described by foregoing summary.As a result are as follows: dry mycelial weight 5.722g/100mL training
Base is supported, mycelia polysaccharide is 669.861mg/100mL culture medium, and exocellular polysaccharide is 1402.065mg/100mL culture medium, and ganoderma lucidum is total
Polysaccharide is 2071.926mg/100mL culture medium.Through detecting, mycotoxin aflatoxin B1, B2 and the deoxidation snow corruption of total starches
Limitation, heavy metal lead, arsenic, mercury, cadmium, the limitation of chromium and pesticide carbendazim, avermectin, the residual of butachlor of sickle-like bacteria enol
Limitation not higher than Limited Doses as defined in GB2761-2017, GB2762-2017, GB2763-2016 or is not detected.
Embodiment two
Using ganoderma lucidum 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 KH2PO40.015g/100mL, MgSO4·
7H2Naturally, adding water to required volume, 121 DEG C of sterilizing 30min shake as culture medium for fermenting by O0.015g/100mL, pH
Bottle sample-loading amount is 40%, inoculum concentration 10%, 24 DEG C of cultivation temperature, revolving speed 150r/min, incubation time 12d.Below by aforementioned
Implement step (3)~(7) in technical solution described by summary of the invention.As a result are as follows: dry mycelial weight 2.4g/100mL culture medium, bacterium
Silk polysaccharide is 158mg/100mL culture medium, and exocellular polysaccharide is 1150.8mg/100mL culture medium, and ganoderma lucidum total starches are
1308.8mg/100mL culture medium；Through detecting, mycotoxin aflatoxin B1, B2 and the deoxynivalenol bacterium alkene of total starches
Limitation, heavy metal lead, arsenic, mercury, cadmium, the limitation of chromium and the pesticide carbendazim of alcohol, avermectin, butachlor residue limits not
Higher than Limited Doses as defined in GB2761-2017, GB2762-2017, GB2763-2016 or it is not detected.
Embodiment three
Using ganoderma lucidum new strains.Wheat bran and rice bran is examined to meet use standard of the invention.Rice bran usage amount is 2.1g/
100mL culture medium, the usage amount of wheat bran are 8.0g/100mL culture medium, add KH2PO40.15g/100mL, MgSO4·7H2O
0.05g/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%, 27 DEG C of cultivation temperature, revolving speed 150r/min, incubation time 6d.Below by aforementioned invention
Implement step (3)~(7) held in described technical solution.As a result are as follows: dry mycelial weight 2.8g/100mL culture medium, mycelia polysaccharide
For 206mg/100mL culture medium, exocellular polysaccharide is 1336.8mg/100mL culture medium, and ganoderma lucidum total starches are 1542.8mg/100mL
Culture medium；Through detecting, the mycotoxin aflatoxin B1 of total starches, the limitation of B2 and deoxynivalenol, a huge sum of money
Belong to lead, arsenic, mercury, cadmium, the limitation of chromium and pesticide carbendazim, avermectin, butachlor residue limits be not higher than GB2761-
2017, it Limited Doses as defined in GB2762-2017, GB2763-2016 or is not detected.
Example IV
Using ganoderma lucidum new strains.Wheat bran and rice bran is examined to meet use standard of the invention.Rice bran usage amount is 1.1g/
100mL culture medium, the usage amount of wheat bran are 9.0g/100mL culture medium, add KH2PO40.2g/100mL, MgSO4·
7H2Naturally, adding water to required volume, 121 DEG C of sterilizing 30min shake as culture medium for fermenting by O0.18g/100mL, pH
Bottle sample-loading amount is 40%, inoculum concentration 10%, 28 DEG C of cultivation temperature, revolving speed 150r/min, incubation time 7d.Aforementioned hair is pressed below
Implement step (3)~(7) in technical solution described by bright content.As a result are as follows: dry mycelial weight 3.3g/100mL culture medium, mycelia
Polysaccharide is 239.4mg/100mL culture medium, and exocellular polysaccharide is 1368.3mg/100mL culture medium, and ganoderma lucidum total starches are
1607.7mg/100mL culture medium；Through detecting, mycotoxin aflatoxin B1, B2 and the deoxynivalenol bacterium alkene of total starches
Limitation, heavy metal lead, arsenic, mercury, cadmium, the limitation of chromium and the pesticide carbendazim of alcohol, avermectin, butachlor residue limits not
Higher than Limited Doses as defined in GB2761-2017, GB2762-2017, GB2763-2016 or it is not detected.
Embodiment five
Using ganoderma lucidum new strains.Wheat bran and rice bran is examined to meet use standard of the invention.Rice bran usage amount is 10g/
100mL culture medium, the usage amount of wheat bran are 0.1g/100mL culture medium, add KH2PO40.05g/100mL, MgSO4·
7H2Naturally, adding water to required volume, 121 DEG C of sterilizing 30min shake as culture medium for fermenting by O0.05g/100mL, pH
Bottle sample-loading amount is 40%, inoculum concentration 10%, 28 DEG C of cultivation temperature, revolving speed 150r/min, incubation time 6d.Aforementioned hair is pressed below
Implement step (3)~(7) in technical solution described by bright content.As a result are as follows: dry mycelial weight 3g/100mL culture medium, mycelia are more
Sugar is 221.9mg/100mL culture medium, and exocellular polysaccharide is 1305.8mg/100mL culture medium, and ganoderma lucidum total starches are 1527.7mg/
100mL culture medium；Through detecting, the mycotoxin aflatoxin B1s 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
Limited Doses as defined in GB2761-2017, GB2762-2017, GB2763-2016 are not detected.
Embodiment six
Using ganoderma lucidum new strains.Wheat bran and rice bran is examined to meet use standard of the invention.Fermentor sample-loading amount is hair
The 75% of fermentation tank volume, fermentation temperature are 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 rice bran 1g/L, wheat bran 100g/L, KH2PO40.15g/L,
MgSO4·7H2O2.5g/L, pH are natural.Implement below by step (3)~(7) in technical solution described by foregoing summary.
As a result are as follows: dry mycelial weight 5.384g/100mL culture medium, mycelia polysaccharide are 602.3mg/100mL culture medium, and exocellular polysaccharide is
1327.8mg/100mL culture medium, ganoderma lucidum total starches are 1930.1mg/100mL culture medium；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 regulation
Limited Doses or be not detected.
Embodiment seven
Using ganoderma lucidum new strains.Wheat bran and rice bran is examined to meet use standard of the invention.Fermentor sample-loading amount is hair
The 75% of fermentation tank volume, fermentation temperature are 24 DEG C, ventilation quantity 1:0.8v/v/mim, mixing speed 150r/min, tank gauge pressure
0.05MPa, inoculum concentration 10%, incubation time 12d, fermentation medium are rice bran 100g/L, wheat bran 1g/L, KH2PO42.5g/L
MgSO4·7H2O0.15g/L, pH are natural.It is real by step (3)~(7) in technical solution described by foregoing summary below
It applies.As a result are as follows: dry mycelial weight 3.2g/100mL culture medium, mycelia polysaccharide are 249.3mg/100mL culture medium, and exocellular polysaccharide is
1323.5mg/100mL culture medium, ganoderma lucidum total starches are 1571.8mg/100mL culture medium；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 regulation
Limited Doses or be not detected.
Embodiment eight
Using ganoderma lucidum new strains.Wheat bran and rice bran is examined to meet use standard of the invention.Fermentor sample-loading amount is hair
The 75% of fermentation tank volume, fermentation temperature are 26 DEG C, ventilation quantity 1:0.8v/v/mim, mixing speed 100r/min, tank gauge pressure
0.05MPa, inoculum concentration 10%, incubation time 7.5d, fermentation medium are rice bran 26g/L, wheat bran 75g/L, KH2PO41.8g/L
MgSO4·7H2O 1.8g/L, pH are natural.It is real by step (3)~(7) in technical solution described by foregoing summary below
It applies.As a result are as follows: dry mycelial weight 2.9g/100mL culture medium, mycelia polysaccharide are 216.5mg/100mL culture medium, and exocellular polysaccharide is
1234.6mg/100mL culture medium, ganoderma lucidum total starches are 1451.1mg/100mL culture medium；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 regulation
Limited Doses or be not detected.
Claims (1)
1. being carried out as steps described below using the method for the ganoderma lucidum new strains production ganoderma lucidum polysaccharide that mutagenesis generates:
(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) rice bran and wheat bran complete feed are directly used in ganoderma strain Ganoderma lucidum as carbon source, nitrogen source and save number
The fermentation medium of CCTCC NO:M 2018908, rice bran concentration are 0.1-10g/100mL, and wheat bran concentration is 0.1-10g/
The total concentration of 100mL, rice bran and wheat bran is 10.1g/100mL, adds potassium dihydrogen phosphate 0.015-0.25g/100mL, seven water sulphur
Sour magnesium 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 rotating speed of agitator 50-150r/min, fermentation time 5-12d；
(3) liquid culture is centrifugally separating to obtain mycelium and fermentation liquid, and measures the yield of exocellular polysaccharide in fermentation liquid；
(4) 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；
(5) 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 ganoderma lucidum polysaccharide product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910608915.3A CN110283861B (en) | 2019-07-05 | 2019-07-05 | Method for producing ganoderan by using ganoderma lucidum strains generated by mutagenesis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910608915.3A CN110283861B (en) | 2019-07-05 | 2019-07-05 | Method for producing ganoderan by using ganoderma lucidum strains generated by mutagenesis |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110283861A true CN110283861A (en) | 2019-09-27 |
| CN110283861B CN110283861B (en) | 2023-05-02 |
Family
ID=68021115
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910608915.3A Active CN110283861B (en) | 2019-07-05 | 2019-07-05 | Method for producing ganoderan by using ganoderma lucidum strains generated by mutagenesis |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110283861B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20010010192A (en) * | 1999-07-16 | 2001-02-05 | 이신영 | Exo-polysaccharide production from submerged mycelial culture of Ganoderma lucidum |
| CN103477994A (en) * | 2013-06-28 | 2014-01-01 | 江苏大学 | Bacterial strain used for producing ganoderma lucidum polysaccharides by complete feed liquid fermentation of rice bran and wheat bran |
| CN105586267A (en) * | 2015-12-25 | 2016-05-18 | 江苏大学 | Ganoderma lucidum mutant strain for production of Ganoderma lucidum mycelia |
| CN105754865A (en) * | 2015-12-25 | 2016-07-13 | 江苏大学 | Method for producing ganoderma lucidum mycelium material by induced mutant strain |
-
2019
- 2019-07-05 CN CN201910608915.3A patent/CN110283861B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20010010192A (en) * | 1999-07-16 | 2001-02-05 | 이신영 | Exo-polysaccharide production from submerged mycelial culture of Ganoderma lucidum |
| CN103477994A (en) * | 2013-06-28 | 2014-01-01 | 江苏大学 | Bacterial strain used for producing ganoderma lucidum polysaccharides by complete feed liquid fermentation of rice bran and wheat bran |
| CN105586267A (en) * | 2015-12-25 | 2016-05-18 | 江苏大学 | Ganoderma lucidum mutant strain for production of Ganoderma lucidum mycelia |
| CN105754865A (en) * | 2015-12-25 | 2016-07-13 | 江苏大学 | Method for producing ganoderma lucidum mycelium material by induced mutant strain |
Non-Patent Citations (2)
| Title |
|---|
| 冯东英 等: "液体发酵高产灵芝多糖培养基的筛选与配方优化", 《食用菌》 * |
| 夏志兰 等: "灵芝深层发酵优良菌株的筛选", 《菌物研究》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110283861B (en) | 2023-05-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Chang et al. | The cultivation and environmental impact of mushrooms | |
| CN103145462B (en) | Enoki mushroom stock culture material and making method thereof | |
| CN106222098B (en) | One plant of monascus strain and its application | |
| CN103477994B (en) | Bacterial strain used for producing ganoderma lucidum polysaccharides by complete feed liquid fermentation of rice bran and wheat bran | |
| CN102876587A (en) | Cordyceps militaris strain for producing cordycepin with high yield | |
| CN102119631B (en) | Grifola frondosa strain for producing polysaccharide with composite raw material of rice bran and wheat bran | |
| CN101991043A (en) | Method for processing compound oat nutritious food with liquid fermentation tricholoma matsutake mycelium polysaccharide | |
| CN103891523B (en) | A kind of Inonotus obliquus artificial cultivation method | |
| CN101463339B (en) | Inocula for preventing apple fungous diseases and increasing yield, and special strain therefore and use thereof | |
| CN102925527A (en) | Method for mixing and fermenting flammulina velutipes and lucid ganoderma | |
| Singh et al. | Characterization and development of cultivation technology of wild split gill Schizophyllum commune mushroom in India | |
| CN102174414B (en) | Application of new morchella costata M8-13 liquid fermentation substance in development of health-care products and medicaments | |
| CN104611236B (en) | Cunninghamella echinulata FAR3 and fermentation thereof are for the method for gamma-linolenic acid oil | |
| CN105586269B (en) | The method for producing maitake mushroom mycelia raw material using mutagenesis Grifola frondosa strain | |
| CN103421861A (en) | Method of manufacturing cordyceps sinensis (Berk.) sacc polysaccharide by liquid state fermentation of rice bran and bran complete feed | |
| CN105754865B (en) | Utilize the method for mutagenic strain production ganoderma lucidum mycelium raw material | |
| CN105586267B (en) | Produce the ganoderma lucidum mutagenic strain of ganoderma lucidum mycelium | |
| CN110527702A (en) | Utilize the method for mutagenic obtained Inonotus obliquus bacterial strain production Fuscoporia obliqua polysaccharide | |
| CN110438010A (en) | One plant of ganoderma strain obtained by ultraviolet mutagenesis | |
| CN110423788A (en) | A method of grifolan is produced using the Grifola frondosa strain that mutagenesis generates | |
| CN110283861A (en) | A method of ganoderma lucidum polysaccharide is produced using the ganoderma strain that mutagenesis generates | |
| CN105586266B (en) | A method of producing selenium-rich Chinese caterpillar fungus bacterial filament raw material | |
| CN110396476A (en) | One plant of Grifola frondosa strain obtained by ultraviolet mutagenesis | |
| CN114181837B (en) | New selenium-enriched Coulomb mushroom strain and artificial cultivation method thereof | |
| CN105754867B (en) | A method of producing selenium-rich grifola frondosus mycelia |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |