CN105586272B - One plant of Ganoderma lucidum bacterial strain and its application - Google Patents
One plant of Ganoderma lucidum bacterial strain and its application Download PDFInfo
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Abstract
The present invention relates to one plant of Ganoderma lucidum bacterial strain and its application, the deposit number of Ganoderma lucidum strain was named NCPSLZ1 of the present invention, bacterial strain NCPSLZ1 are CCTCC No.M2015796；Depositary institution is China typical culture collection center；Preservation date is on December 29th, 2015；Preservation address is Wuhan, China Wuhan University.The mycelium and zymotic fluid of Ganoderma lucidum bacterial strain of the present invention can be used for preparing anti-tumor drug.
Description
Technical field
The present invention provides one plant of Ganoderma lucidum bacterial strain and its application, and in particular to the bacterial strain is in antitumor drug is prepared
Application, belong to microorganism field.
Background technology
Ganoderma lucidum alias 10000 years gill fungus or polyporus lucidus, belong to Basidiomycetes, Aphyllophorales, Polyporaceae, and ganoderma lucidum Pseudomonas is therein
The form of similar deer horn can be formed in Ganoderma lucidum (Ganoderma amboinense) growth, belongs to the rare product in ganoderma lucidum
Kind.Modern medicine research finds that Ganoderma lucidum has enhancing immune, boosts metabolism, antitumor, and the effect such as anti-aging, it is closed
Key active ingredient is ganoderma lucidum polysaccharide and ganodenic acid.
Traditional artificial cultivating ganoderma, cycle length, low production efficiency are affected by environment big.And pass through Ganoderma lucidum submerged fermentation skill
Art, can not only turn out a large amount of mycelium in the short time, shorten the production time, and can obtain the target work that quality is good and stablizes
Property product, nutritive and medicinal value are all not less than fructification, are just progressively being applied to medicine, food, the field such as cosmetics.Spirit
The key of sesame liquid fermentation technology is to find suitable bacterial strain on the one hand to have higher ganoderma lucidum polysaccharide and ganoderma lucidum triterpene
Compound production, on the one hand will adapt to simple cultural method and cheap medium component.
The content of the invention
It is an object of the invention to provide one plant of Ganoderma lucidum bacterial strain and its application, Ganoderma lucidum bacterial strain collection of the present invention is certainly
Tai shan Wild ganoderma, is named as NCPSLZ1, after bacterial strain NCPSLZ1 is by fermentation, the ganoderma lucidum polysaccharide containing high level and
Ganoderma lucidum triterpene compounds.
Ganoderma lucidum polysaccharide has extensive pharmacological activity, can improve immunity of organisms, improves body's hypoxia tolerance, eliminates certainly
By base, suppress tumour, radioresistance, improve liver, marrow, blood synthetic DNA, RNA, protein-energy, extend the service life, ganoderma lucidum is more
Sugar also has stimulation of host non-specific resistance, immune idiosyncrasy and the characteristic for suppressing transplantation tumor physiological activity.To the heart
Vascular diseases, asthma, allergy, neurasthenia, stomach energy etc. have remarkable result.Also there is lowering blood pressure and blood fat, drop blood stasis, improve
Blood circulation, beautifying skin etc. act on.
The pharmacological action of ganoderma lucidum triterpene compounds is：Hepatoprotective effect；Antitumor action；Anti-HIV-1 ` and HIV-1 albumen
Enzymatic activity；Suppress histamine release；Suppress angiotensin converting enzyme (ACE)；Suppress cholesterol biosynthesis；Analgesic activity；It is small to blood
The influence of plate polychlorostyrene；To the inhibitory action of albumen Acacia transesterify enzyme；Antioxidation；Suppress eukaryotic cell dna polymerase
Activity.
The deposit number of Ganoderma lucidum bacterial strain NCPSLZ1 of the present invention is CCTCC No.M2015796；Depositary institution is China
Type Tissue Collection；Preservation date is on December 29th, 2015；Preservation address is Wuhan, China Wuhan University, the deer horn
Ganoderma strain NCPSLZ1 is Ganoderma amboinense NCPSLZ1.
Bacterial strain NCPSLZ1 of the present invention carries out 18sDNA strain idenfications, knot by gold only intelligence biotechnology (Beijing) Co., Ltd
Fruit shows that the bacterial strain belongs to Ganoderma lucidum (Ganoderma amboinense NCPSLZ1), and with published Ganoderma lucidum bacterium
Strain 18sDNA sequences are had any different, and homology is less than 100%.
The method of bacterial strain NCPSLZ1 Liquid Cultures of the present invention, comprises the following steps that：
(1) parent species expand
Take fritter Ganoderma lucidum bacterial strain NCPSLZ1 under aseptic condition, in transposing to slant medium, jump a queue, to constant temperature
In incubator, cultivated 7 days in 25-29 DEG C；Preferably, bacterial strain NCPSLZ1 transposings are cultivated 7 days on slant medium in 27 DEG C；
Slant medium：Potato is cleaned, and peeling, weighs 200g, be cut into small pieces, with 30g wheat bran, add water 1000mL to boil
Boiling, keeps 30min, filters, and agar 20g is added in filtrate, after heating makes it all melt, addition glucose 30g, and biphosphate
Potassium 2g, magnesium sulfate 1g, pH 7.0, is distributed into test tube, is put into high-pressure sterilizing pot sterilizing, and temperature is 121 DEG C, time 30min,
It is spare that test tube slant (18mm × 180mm) is made；
(2) prepared by seed liquor
1cm is taken on slant medium2Mycelium, be inoculated in seed liquid culture medium, be placed in shaking table 25-29 DEG C,
140-160r/min is cultivated 7 days；Preferably, mycelium is inoculated in seed liquid culture medium, is placed in 27 DEG C, 150r/min in shaking table
Culture 7 days
Seed liquid culture medium, as mass fraction：15% murphy juice, 3% wheat bran, 3% glucose, 0.2% biphosphate
Potassium, 0.1% magnesium sulfate, pH 7.0, nutrient solution are sub-packed in 500mL triangular flasks, per bottled 250mL, 121 DEG C of sterilizing 30min,
Taken out after cooling spare.
(3) ferment tank
Seed liquor is inoculated into fermentation tank by 10% (v/v) inoculum concentration, 25-29 DEG C of constant temperature incubation 80h, wherein 0-36 are small
When mycelium lag phase in mixing speed 90-110r/min, mixing speed 110- in mycelium exponential phase when 36-70 is small
Mixing speed 90-110r/min in mycelial growth stationary phase when 130r/min, 70-80 are small；Preferably, mycelium when 0-36 is small
Mixing speed 100r/min in lag phase；Mixing speed 120r/min, 70-80 are small in mycelium exponential phase when 36-70 is small
When mycelial growth stationary phase in mixing speed 100r/min.
Fermentation tank culture medium (mass fraction)：20% murphy juice, 3% wheat bran, 3% glucose, 0.2% biphosphate
Potassium, 0.1% magnesium sulfate, pH 7.0.
The mycelium or zymotic fluid of Ganoderma lucidum bacterial strain of the present invention can be used for preparing anti-tumor drug.
The present invention has the following advantages compared with prior art：
After Ganoderma lucidum bacterial strain NCPSLZ1 of the present invention is by fermentation, ganoderma lucidum polysaccharide and ganodenic acid containing high level
Class compound, available for preparing anti-tumor drug.In the case where not carrying out any optimization, Ganoderma lucidum bacterial strain of the present invention
The extracellular polysaccharide of NCPSLZ1 liquid fermentations, intracellular polyse and triterpene compound content have also respectively reached 4.07g/L, 85mg/
L and 28.8mg/L, higher level is in similar ganoderma strain.After optimization, biomass 12.5g/L, intracellular polyse is
137mg/L, triterpene compound 55mg/L, yield are greatly improved again compared with the shake flask fermentation before being not optimised.
Brief description of the drawings
Fig. 1 is influence of the different carbon source to Ganoderma lucidum mycelial biomass of the present invention；
Fig. 2 is influence of the different nitrogen sources to Ganoderma lucidum mycelial biomass of the present invention；
Fig. 3 is Ganoderma lucidum mycelial growth curve map of the present invention；
Fig. 4 is influences of the initial pH to Ganoderma lucidum mycelial biomass of the present invention；
Fig. 5 is influence of the potato juice concentration to Ganoderma lucidum mycelial biomass of the present invention；
Fig. 6 is influence of the inoculum concentration to Ganoderma lucidum mycelial biomass of the present invention；
Fig. 7 is Ganoderma lucidum mycelium pellet aspect graph of the present invention；
Fig. 8 is hypha form figure of the Ganoderma lucidum of the present invention under Olympus photoelectric microscope AH-2 oil mirrors.
Embodiment
The invention will now be further described with reference to specific embodiments, the advantages and features of the present invention will be with description and
It is apparent.But embodiment is only exemplary, does not form any restrictions to the scope of the present invention.Those skilled in the art should
It should be appreciated that the details and form of technical solution of the present invention can be repaiied without departing from the spirit and scope of the invention
Change or replace, but these modifications and replacement are each fallen within protection scope of the present invention.
1 strain idenfication of embodiment
The deposit number of Ganoderma lucidum bacterial strain NCPSLZ1 of the present invention is CCTCC No.M2015796；Depositary institution is China
Type Tissue Collection；Preservation date is on December 29th, 2015；Preservation address is Wuhan, China Wuhan University.The present invention
The mycelium and zymotic fluid of Ganoderma lucidum bacterial strain can be used for preparing anti-tumor drug.
The method of the bacterial strain NCPSLZ1 Liquid Cultures of the present invention of embodiment 2, comprises the following steps that：
(1) parent species expand
Take fritter Ganoderma lucidum bacterial strain NCPSLZ1 under aseptic condition, in transposing to slant medium, jump a queue, to constant temperature
In incubator, cultivated 7 days in 27 DEG C；
Slant medium：Potato is cleaned, and peeling, weighs 200g, be cut into small pieces, with 30g wheat bran, add water 1000mL to boil
Boiling, keeps 30min, filters, and agar 20g is added in filtrate, after heating makes it all melt, addition glucose 30g, and biphosphate
Potassium 2g, magnesium sulfate 1g, pH 7.0, is distributed into test tube, is put into high-pressure sterilizing pot sterilizing, and temperature is 121 DEG C, time 30min,
It is spare that test tube slant (18mm × 180mm) is made；
(2) prepared by seed liquor
1cm is taken on slant medium2Mycelium, be inoculated in seed liquid culture medium, be placed in shaking table 27 DEG C,
150r/min is cultivated 7 days.
Seed liquid culture medium, as mass fraction：15% murphy juice, 3% wheat bran, 3% glucose, 0.2% biphosphate
Potassium, 0.1% magnesium sulfate, pH 7.0, nutrient solution are sub-packed in 500mL triangular flasks, per bottled 250mL, 121 DEG C of sterilizing 30min,
Taken out after cooling spare.
(3) ferment tank
Seed liquor is inoculated into fermentation tank by 10% (v/v) inoculum concentration, 25-29 DEG C of constant temperature incubation 80h, wherein 0-36 are small
When mycelium lag phase in mixing speed 100r/min, mixing speed 120r/min in mycelium exponential phase when 36-70 is small,
Mixing speed 100r/min in mycelial growth stationary phase when 70-80 is small.
Fermentation tank culture medium (mass fraction)：20% murphy juice, 3% wheat bran, 3% glucose, 0.2% biphosphate
Potassium, 0.1% magnesium sulfate, pH 7.0.
The Ganoderma lucidum bacterial strain NCPSLZ1 production ganoderma lucidum polysaccharide of the present invention of embodiment 3 and triterpene compound capability evaluation
The liquid fermentation medium used during 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 7.0.Inoculum concentration is 5% (V/V),
500mL shaking flasks (dress liquid 250mL), 150r/min, 27 ± 2 DEG C of constant temperature incubation 70h must be assessed with zymotic fluid, further freeze-dried vaccine
Silk powder.
The measure of bacterial strain NCPSLZ1 mycelial biomasses uses dry weight method in the present invention：By zymotic fluid at 4 DEG C, 8000r/
High speed centrifugation 10min under the conditions of min；Precipitation is repeatedly washed with distilled water, is freezed, calculates the biomass (bacterium of ganoderma lucidum mycelium
Filament dry weight g/L zymotic fluids or mycelium dry weight/g), each index determining twice, is averaged.
Ganoderma lucidum polysaccharide is quantitatively detected using phend-sulphuric acid in the present invention：Standard curve is drawn with glucose；Bacterial strain is sent out
Zymotic fluid is concentrated under reduced pressure into 1/5th of original volume, is precipitated with the absolute ethyl alcohol of 4 times of volumes, 12h is stood in 4 DEG C of refrigerators, from
The heart, precipitation plus distilled water dissolving, high speed centrifugation, supernatant freeze after be ganoderma lucidum exocellular polysaccharide powder；1g ganoderma lucidums are freezed into hypha powder
With 50mL distilled water boiling water bath 1h, merge filtrate twice, with 4 times of volumes again with 50mL distilled water, boiling water bath 1h, filtering, precipitation
Absolute ethyl alcohol mixing, 12h is stood in 4 DEG C of refrigerators, is centrifuged, precipitation plus distilled water dissolving, high speed centrifugation, supernatant freezes
Ganoderma lucidum intracellular polyse powder；Use above-mentioned standard curve and polysaccharide powder measure ganoderma polyoses content.
Triterpene compound detection uses vanillic acid-ice acetic acid method：Standard curve is drawn using oleanolic acid；Take 1g bacterium
Silk powder is mixed with 25mL absolute ethyl alcohols, and after ultrasonic 1.5h, filtering, collects filtrate.78 DEG C of water-bath concentrations, are settled to 10mL；Use
The solution of above-mentioned standard curve and institute's constant volume measures triterpene compound content.
As a result：In the case where not carrying out any optimization, Ganoderma lucidum bacterial strain NCPSLZ1 liquid fermentations of the present invention are extracellular
Polysaccharide, intracellular polyse and triterpene compound content have also respectively reached 4.07g/L, 85mg/L and 28.8mg/L, in similar spirit
Higher level is in sesame bacterial strain.
4 liquid fermentation condition of embodiment optimizes
4.1 parent species expand
Potato is cleaned, and peeling, weighs 200g, be cut into small pieces, with 30g wheat bran, add water 1000mL to boil, and keeps 30min,
Filtering, agar 20g is added in filtrate, after heating makes it all melt, adds glucose 30g, potassium dihydrogen phosphate 2g, magnesium sulfate
1g, pH 7.0, is distributed into test tube, is put into high-pressure sterilizing pot sterilizing (121 DEG C of temperature, time 30min), test tube slant is made
(18mm × 180mm) is spare.
Take fritter parent species under aseptic condition, in transposing to slant medium, jump a queue, to constant incubator, 27 DEG C of trainings
Support, after mycelia covers with inclined-plane, will be enlarged by preserving in the 4 DEG C of refrigerators of placement of the parent species after culture stand-by.
It is prepared by 4.2 seed liquors
15% murphy juice, 3% wheat bran, 3% glucose, 0.2% potassium dihydrogen phosphate, 0.1% magnesium sulfate, pH 7.0, culture
Liquid is sub-packed in 500mL triangular flasks, and per bottled 250mL, 121 DEG C of sterilizing 30min, take out spare after cooling.
Middle part of the inoculated by hypha block in slant medium of broad bean size is cut from parent species test tube, 7 are cultivated in 27 ± 2 DEG C
My god.
Lucidum strain after activation is cut into 1cm2The inclined-plane mycelium of size, is inoculated in seed liquid culture medium, is placed in and shakes
Bed in 27 ± 2 DEG C, 150r/min cultivate 7 days.
4.3 trophic factors screen
(15% murphy juice, 3% wheat bran, 3% glucose, 0.2% biphosphate based on the Medium Proportion chosen
Potassium, 0.1% magnesium sulfate, pH 7.0, inquires into influence of the single factors to deep glossy ganoderma fermenting.
To ensure that inoculum concentration is consistent, generation seed liquor culture inoculum concentration is identical with condition of culture.The ganoderma lucidum of bacterium ball is covered with choosing
Fluid nutrient medium, connects 5% seed liquor in screening and culturing medium, is placed in shaking table respectively, rotating speed 150r/min, 27 ± 2 DEG C of constant temperature
70h is cultivated, sees Fig. 7 and Fig. 8.
All unit of weights are all with the Ganoderma lucidum mycelium weight meter of 500mL shaking flasks (dress liquid 250mL) nutrient solution output in experiment
Calculate and handle.
4.3.1 carbon source
It is (beautiful to change the glucose in seed liquor into monose (sorbierite), disaccharide (sucrose, maltose, lactose), polysaccharide respectively
Rice flour, soluble starch), remaining components unchanged.
As a result：As shown in Figure 1, the carbon source that bacterial strain NCPSLZ1 liquid fermentations are most suitable for is glucose, to common in fermentation
Cheap polysaccharide corn flour, soluble starch adaptability are also relatively good, without carbon such as more expensive maltose, lactose, sorbierites
Source.
4.3.2 nitrogen source
Change the wheat bran in seed liquor into inorganic nitrogen (ammonium sulfate), organic nitrogen (yeast extract, analysis for soybean powder), remaining component respectively
It is constant.
As a result：As shown in Fig. 2, the nitrogen source that bacterial strain NCPSLZ1 liquid fermentations are most suitable for is wheat bran, to common honest and clean in fermentation
Valency nitrogen source ammonium sulfate, yeast extract, analysis for soybean powder adaptability are also relatively good, can flexibly be used according to price in fermentation.
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% sulfuric acid
Magnesium, pH 7.0) culture medium is prepared, do culture vessel with 500mL triangular flasks, fill 250mL culture mediums, totally 30 bottles, connect by 10%
Kind amount inoculation, 27 ± 2 DEG C, 150r/min constant temperature incubations, every 6h samplings once, each sample point take 3 bottles, measurement zymotic fluid pH
And dry mycelial weight, common 82h.
As a result：As shown in figure 3, lag phase, when about 0-36 is small, logarithmic phase is followed by stablizing growth when about 36-70 is small
Phase, when most suitable fermentation time is that about 70-80 is small.
4.4.2pH experiment
With culture medium prescription (15% murphy juice, 3% wheat bran, 3% glucose, 0.2% potassium dihydrogen phosphate, 0.1% sulfuric acid
Magnesium) prepare culture medium, before sterilizing with 0.1mol/L NaOH and 0.1mol/L HCl by culture medium be tuned into pH be respectively 3,4,5,
6th, 7,8,9,10,11 totally 9 gradients, are repeated 3 times, observe influences of the different pH to Mycelium Growth of Ganoderma lucidum.
As a result：As shown in figure 4, most suitable initial pH about 7, little with nature (about 6) difference, both can be used.
4.4.3 potato juice concentration
With culture medium prescription (3% wheat bran, 3% glucose, 0.2% potassium dihydrogen phosphate, 0.1% magnesium sulfate, pH 7.0)
Culture medium is prepared, prepares the culture medium of potato juice concentration 5%, 10%, 15%, 20% respectively, remaining condition of culture is constant, repeats
3 times.
As a result：As shown in figure 5, most 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% sulfuric acid
Magnesium, pH 7.0) culture medium is prepared, inoculum concentration is respectively 5%, 10%, 15%, 20%, remaining condition of culture is constant, repeats 3
It is secondary.
As a result：As shown in fig. 6, most suitable inoculum concentration is 10% (V/V) (broth viscosity is excessive during higher).
The preliminary test of 5 ferment tank of embodiment
Production ganoderma lucidum polysaccharide and triterpene compound metering method are shown in 2 part of embodiment, and prepared by seed liquor sees embodiment 3
Point.
Determine that fermentative medium formula is on the basis of embodiment 4：20% murphy juice, 3% wheat bran, 3% glucose,
0.2% potassium dihydrogen phosphate, 0.1% magnesium sulfate, pH.
Feed intake 10L in 16L stainless steel fermentation tanks, inoculum concentration 10%, 27 ± 2 DEG C of constant temperature incubation 80h, bacterium when wherein 0-36 is small
Mixing speed 90-110r/min in filament lag phase, mixing speed 110-130r/ in mycelium exponential phase when 36-70 is small
Mixing speed 90-110r/min in mycelial growth stationary phase when min, 70-80 are small.
As a result：Biomass：12.5g/L intracellular polyse：137mg/L, triterpene compound：55mg/L, yield are relatively not optimised
Preceding shake flask fermentation is greatly improved again.
Claims (8)
1. one plant of Ganoderma lucidum bacterial strain, it is characterised in that Ganoderma lucidum strain was named NCPSLZ1, the preservation of bacterial strain NCPSLZ1
Numbering is CCTCC No.M2015796；Depositary institution is China typical culture collection center；Preservation date is in December, 2015
29 days；Preservation address is Wuhan, China Wuhan University；The bacterial strain can be used for fermenting and producing ganoderma lucidum polysaccharide and triterpene compound, hair
When ferment 80 is small in zymotic fluid intracellular polyse and triterpene compound yield up to 137mg/L and 55mg/L.
2. the method for Ganoderma lucidum bacterial strain NCPSLZ1 Liquid Cultures as claimed in claim 1, comprises the following steps that：
(1) parent species expand
Take fritter Ganoderma lucidum bacterial strain NCPSLZ1 under aseptic condition, in transposing to slant medium, jump a queue, to constant temperature incubation
In case, cultivated 7 days in 25-29 DEG C；
Slant medium：Potato is cleaned, and peeling, weighs 200g, be cut into small pieces, with 30g wheat bran, add water 1000mL to boil, and protects
30min is held, is filtered, agar 20g is added in filtrate, after heating makes it all melt, adds glucose 30g, potassium dihydrogen phosphate 2g,
Magnesium sulfate 1g, pH 7.0, is distributed into test tube, is put into high-pressure sterilizing pot sterilizing, and temperature is 121 DEG C, and time 30min, is made examination
Pipe inclined-plane is spare；
(2) prepared by seed liquor
1cm is taken on slant medium2Mycelium, be inoculated in seed liquid culture medium, be placed in 25-29 DEG C, 140- in shaking table
160r/min is cultivated 7 days；
Seed liquid culture medium, as mass fraction：15% murphy juice, 3% wheat bran, 3% glucose, 0.2% potassium dihydrogen phosphate,
0.1% magnesium sulfate, pH 7.0, nutrient solution are sub-packed in 500mL triangular flasks, per bottled 250mL, 121 DEG C of sterilizing 30min, and cooling
After take out it is spare；
(3) ferment tank
Seed liquor is inoculated into fermentation tank by 10% (v/v) inoculum concentration, 25-29 DEG C of constant temperature incubation 80h, bacterium when wherein 0-36 is small
Mixing speed 90-110r/min in filament lag phase, mixing speed 110-130r/ in mycelium exponential phase when 36-70 is small
Mixing speed 90-110r/min in mycelial growth stationary phase when min, 70-80 are small；
Fermentation tank culture medium, as mass fraction：20% murphy juice, 3% wheat bran, 3% glucose, 0.2% potassium dihydrogen phosphate,
0.1% magnesium sulfate, pH 7.0.
3. the method for Ganoderma lucidum bacterial strain NCPSLZ1 Liquid Cultures according to claim 2, it is characterised in that the step
(1) Ganoderma lucidum bacterial strain NCPSLZ1 transposings are cultivated 7 days on slant medium in 27 DEG C in.
4. the method for Ganoderma lucidum bacterial strain NCPSLZ1 Liquid Cultures according to claim 2, it is characterised in that the step
(2) mycelium is inoculated in seed liquid culture medium in, be placed in shaking table 27 DEG C, 150r/min cultivate 7 days.
5. the method for Ganoderma lucidum bacterial strain NCPSLZ1 Liquid Cultures according to claim 2, it is characterised in that the step
(3) mixing speed 100r/min in mycelium lag phase when 0-36 is small in.
6. the method for Ganoderma lucidum bacterial strain NCPSLZ1 Liquid Cultures according to claim 2, it is characterised in that the step
(3) mixing speed 120r/min in mycelium exponential phase when 36-70 is small in.
7. the method for Ganoderma lucidum bacterial strain NCPSLZ1 Liquid Cultures according to claim 2, it is characterised in that the step
(3) mixing speed 100r/min in mycelial growth stationary phase when 70-80 is small in.
8. the mycelium or zymotic fluid of Ganoderma lucidum bacterial strain as claimed in claim 1 answering in anti-tumor drug is prepared
With.
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| CN112662566B (en) * | 2020-12-15 | 2021-08-31 | 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 |
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