CN105586272A - Ganoderma amboinense strain and application thereof - Google Patents
Ganoderma amboinense strain and application thereof Download PDFInfo
- Publication number
- CN105586272A CN105586272A CN201610058137.1A CN201610058137A CN105586272A CN 105586272 A CN105586272 A CN 105586272A CN 201610058137 A CN201610058137 A CN 201610058137A CN 105586272 A CN105586272 A CN 105586272A
- Authority
- CN
- China
- Prior art keywords
- bacterial strain
- ganoderma lucidum
- ncpslz1
- mycelium
- hour
- 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
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/145—Fungal isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
- A61K36/07—Basidiomycota, e.g. Cryptococcus
- A61K36/074—Ganoderma
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Mycology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Botany (AREA)
- Biochemistry (AREA)
- Medicinal Chemistry (AREA)
- General Engineering & Computer Science (AREA)
- Medical Informatics (AREA)
- Tropical Medicine & Parasitology (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Alternative & Traditional Medicine (AREA)
- Epidemiology (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Medicines Containing Plant Substances (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Mushroom Cultivation (AREA)
Abstract
The invention relates to a Ganoderma amboinense strain and application thereof. The Ganoderma amboinense strain is named NCPSLZ1, and the collection number of the strain NCPSLZ1 is CCTCC No.M2015796; the collection unit is China Center for Type Culture Collection; the collection date is December 29th, 2015; and the collection address is Wuhan University, Wuhan, PRC. The mycelia and fermentation liquid of the Ganoderma amboinense strain can be used for preparing antineoplastic drugs.
Description
Technical field
The invention provides strain Ganoderma lucidum bacterial strain and an application thereof, be specifically related to this bacterial strain and resist swollen in preparationApplication in tumor medicine, belongs to microorganism field.
Background technology
10000 years gill fungus of glossy ganoderma another name or polyporus lucidus, belong to Basidiomycetes, Aphyllophorales, Polyporaceae, glossy ganodermaPseudomonas, can form similar deer horn in the growth of Ganoderma lucidum (Ganodermaamboinense) whereinForm, belongs to the famous and precious kind in glossy ganoderma. Modern medicine research finds that Ganoderma lucidum has the immunity of enhancing, shortEnter metabolism, antitumor, the effect such as delay senility, its crucial active component is GL-B and glossy ganodermaTriterpene.
Traditional Man cultivating ganoderma, the cycle is long, and production efficiency is low, affected by environment large. And pass through glossy ganodermaLiquid fermentation technology, not only can turn out a large amount of mycelium in the short time, shorten the production time, Er QienengObtain quality good and stable targeted activity product, nutrition and medical value are all not less than fructification, just byStep is applied to the fields such as medicine, food, cosmetics. The key of Ganoderma lucidum submerged fermentation technology is to seekLook for applicable bacterial strain, will have higher GL-B and ganoderma lucidum triterpene compounds output, a side on the one handFace is wanted to adapt to simple cultural method and cheap medium component.
Summary of the invention
The object of the present invention is to provide strain Ganoderma lucidum bacterial strain and an application thereof, Ganoderma lucidum bacterium of the present inventionStrain gathers from tai shan Wild ganoderma, called after NCPSLZ1, and bacterial strain NCPSLZ1 passes through after fermentation,The GL-B that contains high level and ganoderma lucidum triterpene compounds.
GL-B has pharmacologically active widely, can improve immunity of organisms, improves body hypoxia-bearing energyPower, eliminates free radical, suppresses tumour, radioresistance, improve liver, marrow, blood synthetic DNA, RNA,Protein ability, life-extending, GL-B also has the non-specific resistance of stimulation of host, immunity is specialThe characteristic of reaction and inhibition transplantation tumor physiologically active. To angiocardiopathy, asthma, allergy, nerveWeak, stomach energy etc. has remarkable result. Also there is hypotensive, reducing blood lipid, fall blood stasis, improve blood circulation,The effects such as beautifying skin.
The pharmacological action of ganoderma lucidum triterpene compounds is: hepatoprotective effect; Antitumor action; Anti-HIV-1 ` andHIV-1 proteinase activity; Suppress histamine release; Angiotensin-converting enzyme inhibition (ACE); Suppress courage solidAlcohol is synthetic; Analgesic activity; On the impact of blood platelet polychlorostyrene; The inhibition of albumen Acacia transesterify enzyme is doneWith; Antioxidation; Suppress eukaryotic cell dna polymerase activity.
The deposit number of Ganoderma lucidum bacterial strain NCPSLZ1 of the present invention is CCTCCNo.M2015796; ProtectHiding unit is Chinese Typical Representative culture collection center; Preservation date is December 29 in 2015Day; Preservation address is Wuhan, China Wuhan University, and this Ganoderma lucidum bacterial strain NCPSLZ1 is GanodermaamboinenseNCPSLZ1。
Bacterial strain NCPSLZ1 of the present invention by gold only intelligence biotechnology (Beijing) Co., Ltd carry out 18sDNABacterial classification qualification, result shows that this bacterial strain belongs to Ganoderma lucidum (GanodermaamboinenseNCPSLZ1),And have any different with published Ganoderma lucidum bacterial strain 18sDNA sequence, homology is lower than 100%.
The method of bacterial strain NCPSLZ1 Liquid Culture of the present invention, concrete steps are as follows:
(1) female kind increased
Under aseptic condition, cut open and get fritter Ganoderma lucidum bacterial strain NCPSLZ1, move and receive on slant medium, addPlug, to constant incubator, cultivates 7 days in 25-29 DEG C; Preferably, bacterial strain NCPSLZ1 moves and receivesOn slant medium, cultivate 7 days in 27 DEG C;
Slant medium: potato is cleaned, peeling, takes 200g, is cut into small pieces, with 30g wheat bran,The 1000mL that adds water boils, and keeps 30min, filters, and adds agar 20g in filtrate, and heating makes them wholeAfter thawing, add glucose 30g, potassium dihydrogen phosphate 2g, magnesium sulfate 1g, pH is 7.0, is distributed into examinationPipe, puts into high-pressure sterilizing pot sterilizing, and temperature is 121 DEG C, and the time is 30min, makes test tube slant(18mm × 180mm) is for subsequent use;
(2) seed liquor preparation
On slant medium, get 1cm2Mycelium, be inoculated in seed liquor culture medium, be placed in shaking table25-29 DEG C, 140-160r/min are cultivated 7 days; Be preferably, mycelium is inoculated in seed liquor culture medium,Be placed in 27 DEG C of shaking tables, 150r/min cultivation 7 days
Seed liquor culture medium, by mass fraction: 15% murphy juice, 3% wheat bran, 3% glucose, 0.2%Potassium dihydrogen phosphate, 0.1% magnesium sulfate, pH is 7.0, nutrient solution is sub-packed in 500mL triangular flask, every bottleDress 250mL, 121 DEG C of sterilizing 30min, cooling rear taking-up is for subsequent use.
(3) ferment tank
Seed liquor is inoculated in fermentation tank by 10% (v/v) inoculum concentration, 25-29 DEG C of constant temperature culture 80h,Wherein mixing speed 90-110r/min in 0-36 hour mycelium lag phase, 36-70 hour mycelium logarithmMixing speed 110-130r/min in growth period, 70-80 hour mycelial growth interior mixing speed stationary phase90-110r/min; Preferably, mixing speed 100r/min in 0-36 hour mycelium lag phase; 36-70 is littleTime mycelium exponential phase in mixing speed 120r/min, in 70-80 hour mycelial growth stationary phase, stirMix speed 100r/min.
Fermentation tank culture medium (mass fraction): 20% murphy juice, 3% wheat bran, 3% glucose, 0.2%Potassium dihydrogen phosphate, 0.1% magnesium sulfate, pH is 7.0.
The mycelium of Ganoderma lucidum bacterial strain of the present invention or zymotic fluid can be used for preparing anti-tumor drug.
The present invention compared with prior art has the following advantages:
Ganoderma lucidum bacterial strain NCPSLZ1 of the present invention by fermentation after, the GL-B that contains high level andGanoderma lucidum triterpene compounds, can be used for preparing anti-tumor drug. In the situation that not carrying out any optimization,Ganoderma lucidum bacterial strain NCPSLZ1 liquid fermentation of the present invention produces exocellular polysaccharide, intracellular polyse and triterpenes chemical combinationThing content has also reached respectively 4.07g/L, 85mg/L and 28.8mg/L, in similar ganoderma strain capable inHigher level. After optimization, biomass is 12.5g/L, and intracellular polyse is 137mg/L, triterpene compoundFor 55mg/L, the shake flask fermentation before output is not optimized is greatly improved again.
Brief description of the drawings
Fig. 1 is the impacts of different carbon sources on Ganoderma lucidum mycelial biomass of the present invention;
Fig. 2 is the impact of different nitrogen sources on Ganoderma lucidum mycelial biomass of the present invention;
Fig. 3 is Ganoderma lucidum mycelial growth curve map of the present invention;
Fig. 4 is the impact of initial pH on Ganoderma lucidum mycelial biomass of the present invention;
Fig. 5 is the impact of murphy juice concentration on Ganoderma lucidum mycelial biomass of the present invention;
Fig. 6 is the impact of inoculum concentration on Ganoderma lucidum mycelial biomass of the present invention;
Fig. 7 is Ganoderma lucidum mycelium pellet aspect graph of the present invention;
Fig. 8 is the mycelia form of Ganoderma lucidum of the present invention under Olympus photoelectric microscope AH-2 oil mirrorFigure.
Detailed description of the invention
Further describe the present invention below in conjunction with specific embodiment, advantage and disadvantage of the present invention will be withDescribe and more clear. But embodiment is only exemplary, do not form any to scope of the present inventionRestriction. It will be understood by those skilled in the art that under without departing from the spirit and scope of the present invention passableDetails to technical solution of the present invention and form are modified or are replaced, but these amendments and replacement all fall intoIn protection scope of the present invention.
Embodiment 1 bacterial classification qualification
The deposit number of Ganoderma lucidum bacterial strain NCPSLZ1 of the present invention is CCTCCNo.M2015796; ProtectHiding unit is Chinese Typical Representative culture collection center; Preservation date is December 29 in 2015Day; Preservation address is Wuhan, China Wuhan University. The mycelium of Ganoderma lucidum bacterial strain of the present invention and zymotic fluidCan be used for preparing anti-tumor drug.
The method of embodiment 2 bacterial strain NCPSLZ1 of the present invention Liquid Culture, concrete steps are as follows:
(1) female kind increased
Under aseptic condition, cut open and get fritter Ganoderma lucidum bacterial strain NCPSLZ1, move and receive on slant medium, addPlug, to constant incubator, cultivates 7 days in 27 DEG C;
Slant medium: potato is cleaned, peeling, takes 200g, is cut into small pieces, with 30g wheat bran,The 1000mL that adds water boils, and keeps 30min, filters, and adds agar 20g in filtrate, and heating makes them wholeAfter thawing, add glucose 30g, potassium dihydrogen phosphate 2g, magnesium sulfate 1g, pH is 7.0, is distributed into examinationPipe, puts into high-pressure sterilizing pot sterilizing, and temperature is 121 DEG C, and the time is 30min, makes test tube slant(18mm × 180mm) is for subsequent use;
(2) seed liquor preparation
On slant medium, get 1cm2Mycelium, be inoculated in seed liquor culture medium, be placed in shaking table27 DEG C, 150r/min are cultivated 7 days.
Seed liquor culture medium, by mass fraction: 15% murphy juice, 3% wheat bran, 3% glucose, 0.2%Potassium dihydrogen phosphate, 0.1% magnesium sulfate, pH is 7.0, nutrient solution is sub-packed in 500mL triangular flask, every bottleDress 250mL, 121 DEG C of sterilizing 30min, cooling rear taking-up is for subsequent use.
(3) ferment tank
Seed liquor is inoculated in fermentation tank by 10% (v/v) inoculum concentration, 25-29 DEG C of constant temperature culture 80h,Wherein mixing speed 100r/min in 0-36 hour mycelium lag phase, 36-70 hour mycelium logarithmic growthMixing speed 120r/min in phase, 70-80 hour mycelial growth interior mixing speed 100r/min stationary phase.
Fermentation tank culture medium (mass fraction): 20% murphy juice, 3% wheat bran, 3% glucose, 0.2%Potassium dihydrogen phosphate, 0.1% magnesium sulfate, pH is 7.0.
Embodiment 3 Ganoderma lucidum bacterial strain of the present invention NCPSLZ1 produces GL-B and triterpene compound energyForce estimation
The liquid fermentation medium using when assessment, the formula (by mass percentage) of culture medium is 15%Murphy juice, 3% wheat bran, 3% glucose, 0.2% potassium dihydrogen phosphate, 0.1% magnesium sulfate, pH is 7.0.Inoculum concentration is 5% (V/V), 500mL shaking flask (dress liquid 250mL), 150r/min, 27 ± 2 DEG C of constant temperature trainingsFoster 70h must assess with zymotic fluid, the further freeze-drying hypha powder that obtains.
In the present invention, the mensuration of bacterial strain NCPSLZ1 mycelial biomass adopts dry weight method: zymotic fluid is existed4 DEG C, high speed centrifugation 10min under 8000r/min condition; Precipitation is repeatedly washed to freeze-drying, meter with distilled waterThe biomass of calculation ganoderma lucidum mycelium (mycelium dry weight g/L zymotic fluid or mycelium dry weight/g), survey by each indexDetermine twice, average.
In the present invention, GL-B quantitatively detects and adopts phenolsulfuric acid method: with glucose drawing standard curve;Bacterial strain fermentation liquor is evaporated to 1/5th of original volume, by the absolute ethyl alcohol precipitation of 4 times of volumes,In 4 DEG C of refrigerators, leave standstill 12h, centrifugal, precipitation adding distil water dissolves, and high speed centrifugation, after supernatant freeze-dryingFor glossy ganoderma exocellular polysaccharide powder; By 1g glossy ganoderma freeze-drying hypha powder and 50mL distilled water, boiling water bath 1h, filters,Precipitation again with 50mL distilled water boiling water bath 1h, merges twice filtrate, mix with the absolute ethyl alcohol of 4 times of volumesClose, leave standstill 12h in 4 DEG C of refrigerators, centrifugal, precipitation adding distil water dissolves, high speed centrifugation, and supernatant freezesDo to obtain glossy ganoderma intracellular polyse powder; Use above-mentioned calibration curve and polysaccharide powder to measure ganoderma polyoses content.
Triterpene compound detects and adopts vanillic acid-ice acetic acid method: use oleanolic acid drawing standard curve;Get 1g hypha powder and mix with 25mL absolute ethyl alcohol, after ultrasonic 1.5h, filter, collect filtrate. 78 DEG C of waterBathe and concentrate, be settled to 10mL; Use the measured in solution triterpene compound of above-mentioned calibration curve and institute's constant volumeContent.
Result: in the situation that not carrying out any optimization, Ganoderma lucidum bacterial strain NCPSLZ1 liquid of the present inventionExocellular polysaccharide is produced in fermentation, and intracellular polyse and triterpene compound content have also reached respectively 4.07g/L,85mg/L and 28.8mg/L, in similar ganoderma strain capable in higher level.
Embodiment 4 liquid fermentation condition optimizations
4.1 female kinds are increased
Potato is cleaned, and peeling, takes 200g, is cut into small pieces, and with 30g wheat bran, 1000mL adds waterBoil, keep 30min, filter, add agar 20g in filtrate, heating adds after it is all meltedGlucose 30g, potassium dihydrogen phosphate 2g, magnesium sulfate 1g, pH is 7.0, is distributed into test tube, puts into high pressureAutoclave sterilizing (121 DEG C of temperature, time 30min), makes test tube slant (18mm × 180mm) standbyWith.
Under aseptic condition, cut open and get the female kind of fritter, move and receive on slant medium, jump a queue, to constant incubatorIn, 27 DEG C of cultivations, after mycelia is covered with inclined-plane, the mother after expansion is cultivated plants to place in 4 DEG C of refrigerators and preservesStand-by.
4.2 seed liquor preparations
15% murphy juice, 3% wheat bran, 3% glucose, 0.2% potassium dihydrogen phosphate, 0.1% magnesium sulfate, pHBe 7.0, nutrient solution is sub-packed in 500mL triangular flask, every bottled 250mL, and 121 DEG C of sterilizing 30min,Cooling rear taking-up is for subsequent use.
Plant test tube and cut the inoculated by hypha block of broad bean size in the middle part of slant medium from mother, in 27 ± 2 DEG CCultivate 7 days.
Lucidum strain after activation is cut into 1cm2The inclined-plane mycelium of size, is inoculated in seed liquor culture mediumIn, be placed in 27 ± 2 DEG C of shaking tables, 150r/min cultivation 7 days.
4.3 trophic factors screenings
Taking the Medium Proportion chosen as basis (15% murphy juice, 3% wheat bran, 3% glucose, 0.2%Potassium dihydrogen phosphate, 0.1% magnesium sulfate, pH is 7.0, inquires into the impact of single factors on deep glossy ganoderma fermenting.
For ensureing that inoculum concentration is consistent, generation seed liquor cultivation inoculum concentration is identical with condition of culture. Bacterium is covered with in choosingThe Ganoderma lucidum by submerged culture base of ball, connects respectively 5% seed liquor in screening and culturing base, is placed in shaking table, rotating speed150r/min, 27 ± 2 DEG C of constant temperature culture 70h, are shown in Fig. 7 and Fig. 8.
In experiment, all unit of weights are all with the glossy ganoderma of 500mL shaking flask (dress liquid 250mL) nutrient solution outputMycelia weight is calculated and is processed.
4.3.1 carbon source
Change respectively the glucose in seed liquor into monose (sorbierite), disaccharide (sucrose, maltose, breastSugar), polysaccharide (corn flour, soluble starch), all the other components unchanged.
Result: as shown in Figure 1, the optimal carbon source of bacterial strain NCPSLZ1 liquid fermentation is glucose, rightCheap polysaccharide corn flour, soluble starch adaptability common in fermentation are also relatively good, more expensive without adoptingThe carbon source such as maltose, lactose, sorbierite.
4.3.2 nitrogenous source
Change respectively the wheat bran in seed liquor into inorganic nitrogen (ammonium sulfate), organic nitrogen (yeast extract, analysis for soybean powder),All the other components unchanged.
Result: as shown in Figure 2, the optimal nitrogenous source of bacterial strain NCPSLZ1 liquid fermentation is wheat bran, to sending outCheap nitrogenous source ammonium sulfate, yeast extract, analysis for soybean powder adaptability common in ferment are also relatively good, can in fermentationUse flexibly according to price.
4.4 other fermentation conditions
4.4.1 ganoderma lucidum mycelium growth curve
With culture medium prescription (15% murphy juice, 3% wheat bran, 3% glucose, 0.2% potassium dihydrogen phosphate,0.1% magnesium sulfate, pH is 7.0) preparation culture medium, do culture vessel with 500mL triangular flask, fill 250mLCulture medium, totally 30 bottles, by 10% inoculum concentration inoculation, 27 ± 2 DEG C, 150r/min constant temperature culture, every6h samples once, and each sample point is got 3 bottles, measures zymotic fluid pH and mycelia dry weight, altogether 82h.
Result: as shown in Figure 3, lag phase is at about 0-36 hour, logarithmic phase is at about 36-70 hour, itsBe the stable growth phase afterwards, the suitableeest fermentation time is about 70-80 hour.
4.4.2pH experiment
With culture medium prescription (15% murphy juice, 3% wheat bran, 3% glucose, 0.2% potassium dihydrogen phosphate,0.1% magnesium sulfate) preparation culture medium, before sterilizing, will cultivate with 0.1mol/LNaOH and 0.1mol/LHClKeynote becomes pH to be respectively 3,4,5,6,7,8,9,10,11 totally 9 gradients, repeats 3 times, seesExamine the impact of different pH on Mycelium Growth of Ganoderma lucidum.
Result: as shown in Figure 4, the suitableeest initial pH approximately 7, little with nature (approximately 6) difference,Both all can use.
4.4.3 murphy juice concentration
With culture medium prescription (3% wheat bran, 3% glucose, 0.2% potassium dihydrogen phosphate, 0.1% magnesium sulfate,PH is 7.0) preparation culture medium, the cultivation of preparing respectively murphy juice concentration 5%, 10%, 15%, 20%Base, all the other condition of culture are constant, repeat 3 times.
Result: as shown in Figure 5, adaptability of soil fermented bean drink concentration is 20%.
4.4.4 inoculum concentration
With culture medium prescription (15% murphy juice, 3% wheat bran, 3% glucose, 0.2% potassium dihydrogen phosphate,0.1% magnesium sulfate, pH is 7.0) preparation culture medium, inoculum concentration is respectively 5%, 10%, 15%, 20%,All the other condition of culture are constant, repeat 3 times.
Result: as shown in Figure 6, the suitableeest inoculum concentration is 10% (V/V) when higher (broth viscosity excessive).
The preliminary test of embodiment 5 ferment tanks
Produce GL-B and triterpene compound metering method and see embodiment 2 parts, enforcement is shown in seed liquor preparationExample 3 parts.
On embodiment 4 bases, determine that fermentative medium formula is: 20% murphy juice, 3% wheat bran, 3%Glucose, 0.2% potassium dihydrogen phosphate, 0.1% magnesium sulfate, pH.
The 10L that feeds intake in 16L stainless steel fermentation tank, 10%, 27 ± 2 DEG C of constant temperature culture 80h of inoculum concentration, whereinMixing speed 90-110r/min in 0-36 hour mycelium lag phase, 36-70 hour mycelium exponential phaseInterior mixing speed 110-130r/min, 70-80 hour mycelial growth interior mixing speed stationary phase90-110r/min。
Result: biomass: 12.5g/L, intracellular polyse: 137mg/L, triterpene compound: 55mg/L,Shake flask fermentation before output is not optimized is greatly improved again.
Claims (8)
1. a strain Ganoderma lucidum bacterial strain, is characterized in that, Ganoderma lucidum bacterial strain called after NCPSLZ1,The deposit number of bacterial strain NCPSLZ1 is CCTCCNo.M2015796; Depositary institution is Chinese Typical Representative trainingSupport thing preservation center; Preservation date is on December 29th, 2015; Preservation address is that Wuhan, China Wuhan is largeLearn.
2. the method for Ganoderma lucidum bacterial strain NCPSLZ1 Liquid Culture as claimed in claim 1, specifically stepRapid as follows:
(1) female kind increased
Under aseptic condition, cut open and get fritter Ganoderma lucidum bacterial strain NCPSLZ1, move and receive on slant medium, addPlug, to constant incubator, cultivates 7 days in 25-29 DEG C;
Slant medium: potato is cleaned, peeling, takes 200g, is cut into small pieces, with 30g wheat bran,The 1000mL that adds water boils, and keeps 30min, filters, and adds agar 20g in filtrate, and heating makes them wholeAfter thawing, add glucose 30g, potassium dihydrogen phosphate 2g, magnesium sulfate 1g, pH is 7.0, is distributed into examinationPipe, puts into high-pressure sterilizing pot sterilizing, and temperature is 121 DEG C, and the time is 30min, makes test tube slant for subsequent use;
(2) seed liquor preparation
On slant medium, get 1cm2Mycelium, be inoculated in seed liquor culture medium, be placed in shaking table25-29 DEG C, 140-160r/min are cultivated 7 days;
Seed liquor culture medium, by mass fraction: 15% murphy juice, 3% wheat bran, 3% glucose, 0.2%Potassium dihydrogen phosphate, 0.1% magnesium sulfate, pH is 7.0, nutrient solution is sub-packed in 500mL triangular flask, every bottleDress 250mL, 121 DEG C of sterilizing 30min, cooling rear taking-up is for subsequent use;
(3) ferment tank
Seed liquor is inoculated in fermentation tank by 10% (v/v) inoculum concentration, 25-29 DEG C of constant temperature culture 80h,Wherein mixing speed 90-110r/min in 0-36 hour mycelium lag phase, 36-70 hour mycelium logarithmMixing speed 110-130r/min in growth period, 70-80 hour mycelial growth interior mixing speed stationary phase90-110r/min；
Fermentation tank culture medium, by mass fraction: 20% murphy juice, 3% wheat bran, 3% glucose, 0.2%Potassium dihydrogen phosphate, 0.1% magnesium sulfate, pH is 7.0.
3. the method for Ganoderma lucidum bacterial strain NCPSLZ1 Liquid Culture according to claim 2, its spyLevy and be, in described step (1), Ganoderma lucidum bacterial strain NCPSLZ1 moves and receives on slant medium, 27 DEG CMiddle cultivation 7 days.
4. the method for Ganoderma lucidum bacterial strain NCPSLZ1 Liquid Culture according to claim 2, its spyLevy and be, mycelium is inoculated in seed liquor culture medium in described step (2), be placed in 27 DEG C of shaking tables,150r/min cultivates 7 days.
5. the method for Ganoderma lucidum bacterial strain NCPSLZ1 Liquid Culture according to claim 2, its spyLevy and be, mixing speed 100r/min in 0-36 hour mycelium lag phase in described step (3).
6. the method for Ganoderma lucidum bacterial strain NCPSLZ1 Liquid Culture according to claim 2, its spyLevy and be, mixing speed 120r/min in 36-70 hour mycelium exponential phase in described step (3).
7. the method for Ganoderma lucidum bacterial strain NCPSLZ1 Liquid Culture according to claim 2, its spyLevy and be, 70-80 hour mycelial growth interior mixing speed 100r/min stationary phase in described step (3).
8. the mycelium of Ganoderma lucidum bacterial strain or zymotic fluid are antineoplastic in preparation as claimed in claim 1Application in medicine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610058137.1A CN105586272B (en) | 2016-01-28 | 2016-01-28 | One plant of Ganoderma lucidum bacterial strain and its application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610058137.1A CN105586272B (en) | 2016-01-28 | 2016-01-28 | One plant of Ganoderma lucidum bacterial strain and its application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105586272A true CN105586272A (en) | 2016-05-18 |
| CN105586272B CN105586272B (en) | 2018-04-20 |
Family
ID=55926180
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610058137.1A Active CN105586272B (en) | 2016-01-28 | 2016-01-28 | One plant of Ganoderma lucidum bacterial strain and its application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105586272B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107603886A (en) * | 2017-09-29 | 2018-01-19 | 福建绿宝食品集团有限公司 | A kind of preparation method of Ganoderma lucidum liquid spawn |
| CN112662566A (en) * | 2020-12-15 | 2021-04-16 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Ganoderma lucidum spore-less variety with high yield of polysaccharide and artificial cultivation method thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102726665A (en) * | 2012-06-13 | 2012-10-17 | 山东省农业科学院农产品研究所 | Method for producing corn flour by the solid state fermentation of maize pulp by using edible fungi |
-
2016
- 2016-01-28 CN CN201610058137.1A patent/CN105586272B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102726665A (en) * | 2012-06-13 | 2012-10-17 | 山东省农业科学院农产品研究所 | Method for producing corn flour by the solid state fermentation of maize pulp by using edible fungi |
Non-Patent Citations (2)
| Title |
|---|
| 汤坤鹏: "灰色关联度法筛选适合液体深层发酵灵芝菌株", 《核农学报》 * |
| 鉏晓艳: "灵芝真菌的液体深层发酵技术", 《饲料工业》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107603886A (en) * | 2017-09-29 | 2018-01-19 | 福建绿宝食品集团有限公司 | A kind of preparation method of Ganoderma lucidum liquid spawn |
| CN112662566A (en) * | 2020-12-15 | 2021-04-16 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Ganoderma lucidum spore-less variety with high yield of polysaccharide and artificial cultivation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105586272B (en) | 2018-04-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105586273B (en) | A kind of Ganoderma lucidum bacterium solution fermentation culture method | |
| CN100432212C (en) | Grifola frondosa strain, culture method and application thereof | |
| CN106222098B (en) | One plant of monascus strain and its application | |
| CN103343152B (en) | The method of polysaccharide produced by a kind of Grifola frondosa mutagenic fungi liquid state fermentation rice bran wheat bran complete feed | |
| CN102119631B (en) | Grifola frondosa strain for producing polysaccharide with composite raw material of rice bran and wheat bran | |
| CN102864111B (en) | Schizochytrium limacinum strain for producing docosahexaenoic acid | |
| CN103477994B (en) | Bacterial strain used for producing ganoderma lucidum polysaccharides by complete feed liquid fermentation of rice bran and wheat bran | |
| CN108410744B (en) | Fusion strain for producing polysaccharide, adenosine and cordycepin | |
| CN102895685B (en) | Sterilization system for medium, sterilization method using same, and culture method for cordyceps militaris | |
| CN109234331A (en) | A kind of Ganoderma lucidum submerged fermentation is naturally cultivated and tunning complete utilization method | |
| CN105969673A (en) | Fermentation method for increasing yield of haematochrome and uranidin in monascus purpureus | |
| CN103190576A (en) | Cordyceps militaris soybean and preparation method thereof | |
| CN102925527A (en) | Method for mixing and fermenting flammulina velutipes and lucid ganoderma | |
| CN104087512A (en) | Mortierella alpine producing polyunsaturated fatty acids and application thereof | |
| CN102816806A (en) | Production method of grifolan selenium compound | |
| CN102816701B (en) | Strain used for fermenting rice bran and wheat bran extracts for producing grifolan | |
| CN1450168A (en) | Process for raising glutathion yield by fermentation of tornla yeast | |
| CN103525748B (en) | The chlamydosporic production method of trichoderma harziarum | |
| CN102766663B (en) | Preparation method of active polysaccharides from phellinus linteus | |
| CN108410854B (en) | Method for preparing cordyceps militaris and morchella fused strain | |
| CN104911135B (en) | A kind of trehalose synthase production bacterial strain and its application | |
| CN105441334B (en) | Produce bacterial strain and its application of grifolan | |
| CN105586272A (en) | Ganoderma amboinense strain and application thereof | |
| CN103060210B (en) | Polymorphic hansenula polymorpha mutant strain and application of polymorphic hansenula polymorpha mutant strain in glutathione biosynthesis | |
| CN101302480B (en) | High yield gamma-reanal monascus ruber Mr-5 bacterial strain, screening method and use thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |