CN107916230A - A kind of Ganoderma lucidum mycelium zymotic fluid of high ganoderma polyoses content and preparation method thereof - Google Patents
A kind of Ganoderma lucidum mycelium zymotic fluid of high ganoderma polyoses content and preparation method thereof Download PDFInfo
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- CN107916230A CN107916230A CN201711249253.2A CN201711249253A CN107916230A CN 107916230 A CN107916230 A CN 107916230A CN 201711249253 A CN201711249253 A CN 201711249253A CN 107916230 A CN107916230 A CN 107916230A
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Abstract
The present invention relates to fungi fermentation field, and in particular to a kind of Ganoderma lucidum mycelium zymotic fluid of high ganoderma polyoses content and preparation method thereof, this method includes：Ganoderma strain is obtained into mycelia block after slant tube culture, takes the inoculated by hypha block to be transferred to malt medium after carrying out cultural hypha to plating medium, 25~29 DEG C of liquid fermentation and cultures 3~7 days.This method uses mycelium liquid fermentation method, and ganoderma polyoses content is high, and fermentation process is more environmentally friendly, and cost is lower, and required time is shorter, and fermentation process is simple, easy to operation, is adapted to large-scale industrial production, convenient to promote.
Description
Technical field
The present invention relates to fungi fermentation field, ferments in particular to a kind of Ganoderma lucidum mycelium of high ganoderma polyoses content
Liquid and preparation method thereof.
Background technology
Ganoderma lucidum Ganoderma lucidum [(Leyss.ex.Fr.) Karst] belong to Basidiomycetes, Polyporaceae, ganoderma lucidum
Belong to higher fungus.Ganoderma lucidum has the history of more than one thousand years in China, early in《Sheng Nong's herbal classic》In just it is on the books, ganoderma lucidum consolidates training
Member, eats not old, macrobiosis of making light of one's life by commiting suicide long.《Compendium of Materia Medica》It is classified as top grade medicine, it is indicated that and red sesame " it is bitter, put down, it is nontoxic, cure mainly in the heart
Knot, invigorating heart gas, bowl spares, increases wisdom, does not forget ", purple sesame " nontoxic, good color, controls consumptive disease, control hemorrhoid ".
Mainly there is contained active material in ganoderma lucidum：Polysaccharide, triterpenes, ucleosides, sterols, alkaloids, furans
Derivative, amino acid polypeptide class, trace element, aliphatic acid etc., wherein, ganoderma lucidum polysaccharide is the main bioactive ingredients of ganoderma lucidum.
Ganoderma lucidum polysaccharide has many pharmacological activity：Immunity of organisms can be improved, accelerates blood microcirculation, improves blood oxygen ability,
Invalid oxygen demand under reduction body inactive state, elimination interior free yl, the closed stratum of raising body cell film, anti-radiation,
Liver, marrow, blood synthetic DNA, RNA, the ability of protein are improved, extends service life etc..
Wild ganoderma is very rare in nature, and wherein ganoderma polyoses content is not high enough, make exploitation ganoderma lucidum product into
This is excessive.Secondly the content that Ganoderma Lucidum cycle is long, extracts polysaccharide is relatively low, and product quality is easily by amblent air temperature and insect pest
Influence, it has not been convenient to promote.It is raw material that the ganoderma active such as ganoderma lucidum polysaccharide material production at present, which relies primarily on artificial cultivation fructification,
Obtained in the method for extraction.Publication number CN105294875A, publication date are that the application documents of on 2 3rd, 2016 disclose one kind
The method that ganoderma lucidum polysaccharide is extracted from ganoderma lucidum fruitbody, comprises the following steps：Ganoderma lucidum crushes, sodium chlorite-glacial acetic acid system oxygen
Change is handled, and residue adds water ultrasonic extraction, and supernatant centrifuges to obtain Thick many candies through alcohol precipitation.The invention utilizes sodium chlorite-glacial acetic acid body
It is to clear up the lignin and cellulose in ganoderma lucidum fruitbody, polysaccharide therein is more discharged and is extracted with ultrasonic wave added
Combination is taken to extract polysaccharide.The yield of this method ganoderma lucidum polysaccharide is 13.51%.But this method needs to obtain ganoderma lucidum in fact
Body, production cycle length, cost are higher.Need first to extract Linzhi's Thick many candies at the same time, then purify, complex steps, and can produce very
More byproducts, environment easy to pollute.
Publication number CN106431699A, publication date are that the application documents of on 2 22nd, 2017 disclose raising ganoderma lucidum polysaccharide
The culture medium of content, comprises the following steps：Walnut shell is a, and soybean stalk is a, and rice bran is a, and corn flour is a, dregs of beans one
Part, Chinese medicine additive is a, and sucrose is a, and land plaster is a, and precipitated calcium carbonate is a；The Chinese medicine additive is by following heavy
The raw material of amount part is made：Cattail pollen is a, and palm fibre puts portion, and the root of purple-flowered peucedanum is a, and the vine of multiflower knotweed is a, and muskmelon pedicel is a.This method prepares culture
The method and step of base is cumbersome, its culture medium is the culture medium of cultured mycelia, and materials are too inconvenient, are not easy industrialized production.
Publication number CN105017441A, publication date are that the application documents of on 2 22nd, 2017 disclose a kind of ganoderma lucidum polysaccharide
Extracting method, comprise the following steps：Ganoderma lucidum ethanol is extracted, takes filter residue spare；Filter residue water is carried, it is spare to collect filtrate；Will
Filtrate concentrated extract, ethanol precipitation, centrifuging to precipitate；Precipitation successively absolute ethyl alcohol, acetone is washed respectively, will be heavy after washing
Shallow lake is dissolved with distilled water, is crossed the hollow-fibre membrane that molecular cut off is 50,000, is collected concentrate, concentrate then is crossed retention point
Son amount is 80,000 hollow-fibre membrane, collects permeate centrifugation, filtering, takes the spare past filtrate of filtrate to add activated carbon decolorizing, mistake
Filter, takes filtrate to be spray-dried, and is ganoderma lucidum polysaccharide.Its obtained polyoses content is 92.3%.This method step is complicated, and material holds
Environment easy to pollute is, it is necessary to obtain ganoderma lucidum fruitbody, it is necessary to which time length, of high cost.
Publication number CN103073651A, publication date are that the application documents of on 2 22nd, 2017 disclose a kind of ganoderma lucidum polysaccharide
Extracting method, comprise the following steps：The present invention discloses a kind of ganoderma lucidum polysaccharide method and its application.The present invention uses ultrasonic wave knot
Enzymatic Extraction ganoderma lucidum polysaccharide is closed, after ganoderma lucidum crushes, is separated by extraction, ultrafiltration, concentration and ethanol precipitation and obtains ganoderma lucidum polysaccharide.Spirit
Sesame polysaccharide extract rate is 3.14-4.21%.This method also needs to obtain ganoderma lucidum fruitbody, it is necessary to which the time is grown, and step is complicated, no
Easy large-scale production.
In view of this, it is special to propose the present invention.
The content of the invention
It is an object of the invention to provide a kind of preparation method of the Ganoderma lucidum mycelium zymotic fluid of high ganoderma polyoses content, with solution
Certainly the ganoderma lucidum cultural method production cycle is longer, it is necessary to which adopted son's entity, cost is higher in the prior art, and ganoderma lucidum polysaccharide yield is not
Active ingredient composition and content have the problems such as larger difference between enough high, large-scale production difficulties and different batches.
In order to realize the above-mentioned purpose of the present invention, spy uses following technical scheme：
According to an aspect of the present invention, the present invention relates to a kind of preparation of the Ganoderma lucidum mycelium zymotic fluid of high ganoderma polyoses content
Method, including：
Ganoderma strain is obtained into mycelia block after slant tube culture, takes the inoculated by hypha block to be carried out to plating medium
It is transferred to malt medium after cultural hypha, 25~29 DEG C of liquid fermentation and cultures 3~7 days.
Ganoderma lucidum polysaccharide was both present in mycelial cell, can also secrete in extracellular, therefore can be from ganoderma lucidum fruitbody, spore
Separation and Extraction in son, can also extract from the mycelium of liquid fermentation.The prior art usually cultivates ganoderma lucidum fruitbody to obtain
Ganoderma lucidum polysaccharide, but time-consuming for this method, is not suitable for large-scale production, and it is present in the ganoderma lucidum polysaccharide in fructification in extraction
More difficult exudation.
Ganoderma lucidum mycelium growth cycle is short, from mycelium extract polysaccharide have save ground, it is time saving, laborsaving, can intensive metaplasia
The advantages that producing polysaccharide.After ganoderma lucidum is carried out liquid fermentation, directly zymotic fluid can be concentrated, the conventional technical means such as alcohol precipitation is from fermentation
Thick many candies are obtained in liquid or mycelium therein, it is more simple that extraction process compares fructification extraction.
The invention further relates to the Ganoderma lucidum mycelium fermentation for the high ganoderma polyoses content being prepared by method as described above
Liquid.
Compared with prior art, beneficial effects of the present invention are：
It is side of the raw material to extract that the ganoderma active such as ganoderma lucidum polysaccharide material production at present, which relies primarily on artificial cultivation fructification,
Method obtains, contained active substance higher in Ganoderma lucidum mycelium zymotic fluid of the present invention, and the liquid fermentation time
Compared with it is short, possibility of pollution is small, be more advantageous to ganoderma lucidum polysaccharide industrial expansion.It is resulting by preferred culture medium and incubation time
Ganoderma polyoses content it is very high.
Embodiment
The present invention relates to a kind of preparation method of the Ganoderma lucidum mycelium zymotic fluid of high ganoderma polyoses content, including：
Ganoderma strain is obtained into mycelia block after slant tube culture, takes the inoculated by hypha block to be carried out to plating medium
It is transferred to malt medium after cultural hypha, 25~29 DEG C of liquid fermentation and cultures 3~7 days.
Preferably, when the ganoderma strain is U.S. ganoderma lucidum, incubation time is 3 days；When the ganoderma strain is deer horn
During ganoderma lucidum, incubation time is 4 days；When the ganoderma strain is Chinese red sesame, incubation time is 6 days；When the ganoderma strain
For black sesame when, incubation time be 4 days.
This method uses mycelium liquid fermentation method, and ganoderma polyoses content is high, and fermentation process is more environmentally friendly, and cost is lower, institute
Take time shorter, fermentation process is simple, easy to operation, is adapted to large-scale industrial production, convenient to promote.
Preferably, the preparation method of the Ganoderma lucidum mycelium zymotic fluid of high ganoderma polyoses content as described above, the tablet training
Supporting base includes：
25~35g/L of malt extract, 2~4g/L of soy peptone, 2~4w/v% of agar malt extract, soy peptone
0.2~0.4w/v%, 1.3~1.7w/v% of agar, solvent are water, pH5.4~5.8；
It is furthermore preferred that the preparation method of the Ganoderma lucidum mycelium zymotic fluid of high ganoderma polyoses content as described above, the tablet
Culture medium includes：
It is 27~33g/L of malt extract, 2.5~3.5g/L of soy peptone, 2.5~3.5w/v% of agar malt extract, big
Legumin 0.25~0.35w/v% of peptone, 1.4~1.6w/v% of agar, solvent are water, pH5.5~5.7；
Most preferably, the plating medium includes：
Malt extract 30g/L, soy peptone 3g/L, agar malt extract 3w/v%, soy peptone 0.3w/v%, fine jade
Fat 1.5w/v%, solvent are water, pH5.6.
Preferably, the preparation method of the Ganoderma lucidum mycelium zymotic fluid of high ganoderma polyoses content as described above, takes the mycelia
The inoculum concentration that block is inoculated into plating medium is each 0.7~1.3cm of plating2Mycelia block.
Preferably, the preparation method of the Ganoderma lucidum mycelium zymotic fluid of high ganoderma polyoses content as described above, the mycelia training
Foster condition of culture is cultivated 6~8 days for 28~32 DEG C；
It is furthermore preferred that the condition of culture of the cultural hypha is cultivated 7 days for 30 DEG C.
Preferably, the preparation method of the Ganoderma lucidum mycelium zymotic fluid of high ganoderma polyoses content as described above, the malt training
Supporting base includes：
1.5~2.5w/v% of fructus hordei germinatus leaching powder, 1.5~2.5w/v% of glucose, 0.07~0.13w/v% of peptone, yeast
0.07~0.13w/v% of cream, solvent are water；
It is furthermore preferred that the preparation method of the Ganoderma lucidum mycelium zymotic fluid of high ganoderma polyoses content as described above, the malt
Culture medium includes：
1.7~2.3w/v% of fructus hordei germinatus leaching powder, 1.7~2.3w/v% of glucose, 0.09~0.11w/v% of peptone, yeast
0.09~0.11w/v% of cream, solvent are water；
Most preferably, the malt medium includes：
Fructus hordei germinatus leaching powder 2w/v%, glucose 2w/v%, peptone 0.1w/v%, yeast extract 0.1w/v%, solvent are water.
Preferably, the preparation method of the Ganoderma lucidum mycelium zymotic fluid of high ganoderma polyoses content as described above, is transferred to malt
Inoculum concentration when culture medium carries out liquid fermentation culture is per 1.3~1.7cm of 100ml culture medium inoculateds2Mycelia block.
Preferably, the preparation method of the Ganoderma lucidum mycelium zymotic fluid of high ganoderma polyoses content as described above, the Ganoderma Lucidum
Strain includes any of U.S. ganoderma lucidum, Chinese red sesame, Ganoderma lucidum and black sesame；
It is furthermore preferred that the ganoderma strain is selected from U.S. ganoderma lucidum or Ganoderma lucidum；Most preferably Ganoderma lucidum.
The invention further relates to the Ganoderma lucidum mycelium fermentation for the high ganoderma polyoses content being prepared by method as described above
Liquid.
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the invention.It is not specified in embodiment specific
Condition person, the condition suggested according to normal condition or manufacturer carry out.Reagents or instruments used without specified manufacturer, is
Can be with conventional products that are commercially available.
Embodiment
1st, the preparation of culture medium and solution
Plating medium：MEA (fructus hordei germinatus leaching powder agar medium Malt Extract Ager)：Malt extract 30g, soybean
Peptone 3g, agar malt extract 3w/v%, soy peptone 0.3w/v%, agar 1.5w/v%, H2O 1L constant volumes, pH5.6,
121 DEG C of sterilizing 15min.
Malt medium：Fructus hordei germinatus leaching powder 2w/v%, glucose 2w/v%, peptone 0.1w/v%, yeast extract 0.1w/v%,
H2O 1L constant volumes, 115 DEG C of sterilizing 20min.
CYM culture mediums：Glucose 20g, peptone 2g, dusty yeast 2g, MgSO4﹒ 7H2O0.5g、K2HPO4 1g、KH2PO4
0.46g, H2O 1L。
2. ganoderma strain liquid fermentation culturing method
Each bacterial strain takes 1 piece of 1cm after slant tube culture2Inoculated by hypha block, in 30 DEG C, is cultivated 7 days to plating medium
Afterwards.Then 1.5cm is taken from plating medium2Be inoculated into respectively malt medium (300ml conical flasks, liquid amount 100ml) and
CYM culture mediums, in 27 DEG C, after cultivating three days, since the 4th day, while culture, while record.
3rd, during ganoderma strain liquid fermentation polysaccharide yield measure
Liquid 1ml is sampled in 50ml centrifuge tubes, adds 95% alcohol chromatography 24h of 3 times of volumes or so；5000r/min, from
Heart 20min, abandons supernatant；Dry 2-3h will be deposited in cryogenic temperature freezing drying machine；Again plus 20ml ultra-pure waters dissolve sediment
Suitable concentration is diluted to, fully after dissolving, takes 1ml to be measured as prepare liquid, its polyoses content with phend-sulphuric acid.
Phend-sulphuric acid assay method：Take 1ml prepare liquids to be added in test tube, then add 1.0ml into test tube thereto
5% phenol solution, be then quickly added into the 5.0ml concentrated sulfuric acids (it is vertical with liquid level to add, do not contact test tube wall, so as to reaction solution
It is sufficiently mixed), reaction 30min is stood after vibration, 490nm surveys absorbance, then calculates prepare liquid by glucose mark song and obtain polysaccharide
Content.
4th, the screening of species of fermenting fermentation number of days and ganoderma lucidum species
(1), the selection of ganoderma lucidum species and the number of days that ferments during ganoderma lucidum fruitbody ferments
Ganoderma lucidum species is enriched, and the chemical composition of the ganoderma lucidum of different cultivars is also different.Conventional selection is Black Ganoderma
Entity zymotic fluid.Nowadays we select U.S. ganoderma lucidum, Chinese red sesame, Ganoderma lucidum.
Selection U.S. ganoderma lucidum, Chinese red sesame, Ganoderma lucidum, black sesame, using the content of polysaccharide in fermentation liquid as Testing index,
Screening more suitably ganoderma lucidum species and fermentation number of days.1.5cm is taken from mycelial plating medium2Liquid training is inoculated into respectively
Base and CYM culture mediums are supported, in 27 DEG C, after cultivating three days, since the 4th day, culture, and meanwhile record, until the 7th day.
As following table understands that the polyoses content in mycelium fermentation broth in malt medium is than more in CYM culture medium
Sugared content is high.So malt medium is best medium.In malt medium, wherein fermentation of the Ganoderma lucidum at the 4th day
Polyoses content highest in liquid, extraction polysaccharide effect is best, and polyoses content is up to 3.615mg/ml.
1 mycelium fermentation broth testing result of table
Finally it should be noted that：The above embodiments are only used to illustrate the technical solution of the present invention., rather than its limitations；To the greatest extent
Pipe is described in detail the present invention with reference to foregoing embodiments, but it will be understood by those of ordinary skill in the art that：Its
It can still modify to the technical solution described in foregoing embodiments, either to which part or all technical characteristic
Carry out equivalent substitution；And these modifications or replacement, the essence of appropriate technical solution is departed from various embodiments of the present invention skill
The scope of art scheme.
Claims (10)
- A kind of 1. preparation method of the Ganoderma lucidum mycelium zymotic fluid of high ganoderma polyoses content, it is characterised in that including：Ganoderma strain is obtained into mycelia block after slant tube culture, takes the inoculated by hypha block to carry out mycelia to plating medium It is transferred to malt medium after culture, 25~29 DEG C of liquid fermentation and cultures 3~7 days.
- 2. the preparation method of the Ganoderma lucidum mycelium zymotic fluid of high ganoderma polyoses content according to claim 1, it is characterised in that The plating medium includes：25~35g/L of malt extract, 2~4g/L of soy peptone, 2~4w/v% of agar malt extract, soy peptone 0.2~ 0.4w/v%, 1.3~1.7w/v% of agar, solvent are water, pH5.4~5.8.
- 3. the preparation method of the Ganoderma lucidum mycelium zymotic fluid of high ganoderma polyoses content according to claim 2, it is characterised in that The plating medium includes：27~33g/L of malt extract, 2.5~3.5g/L of soy peptone, 2.5~3.5w/v% of agar malt extract, soybean egg White 0.25~0.35w/v% of peptone, 1.4~1.6w/v% of agar, solvent are water, pH5.5~5.7.
- 4. the preparation method of the Ganoderma lucidum mycelium zymotic fluid of the high ganoderma polyoses content according to Claims 2 or 3, its feature exist In the inoculum concentration for taking the inoculated by hypha block to plating medium is each 0.7~1.3cm of plating2Mycelia block.
- 5. the preparation method of the Ganoderma lucidum mycelium zymotic fluid of the high ganoderma polyoses content according to Claims 2 or 3, its feature exist In the condition of culture of the cultural hypha is cultivated 6~8 days for 28~32 DEG C.
- 6. the preparation method of the Ganoderma lucidum mycelium zymotic fluid of high ganoderma polyoses content according to claim 1, it is characterised in that The malt medium includes：1.5~2.5w/v% of fructus hordei germinatus leaching powder, 1.5~2.5w/v% of glucose, 0.07~0.13w/v% of peptone, yeast extract 0.07~0.13w/v%, solvent are water.
- 7. the preparation method of the Ganoderma lucidum mycelium zymotic fluid of high ganoderma polyoses content according to claim 6, it is characterised in that The malt medium includes：1.7~2.3w/v% of fructus hordei germinatus leaching powder, 1.7~2.3w/v% of glucose, 0.09~0.11w/v% of peptone, yeast extract 0.09~0.11w/v%, solvent are water.
- 8. the preparation method of the Ganoderma lucidum mycelium zymotic fluid of the high ganoderma polyoses content according to claim 6 or 7, its feature exist In the inoculum concentration being transferred to when malt medium carries out liquid fermentation culture is per 1.3~1.7cm of 100ml culture medium inoculateds2's Mycelia block.
- 9. the preparation method of the Ganoderma lucidum mycelium zymotic fluid of high ganoderma polyoses content according to claim 1, it is characterised in that The ganoderma strain includes any of U.S. ganoderma lucidum, Chinese red sesame, Ganoderma lucidum and black sesame.
- 10. the Ganoderma lucidum mycelium zymotic fluid for the high ganoderma polyoses content that claim 1~9 any one of them method is prepared.
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| CN109777847A (en) * | 2019-03-01 | 2019-05-21 | 宁德师范学院 | The method of body cell is affine ganoderma strain to common fermentation production strong anti-oxidative activity polysaccharide |
| CN110903982A (en) * | 2019-11-14 | 2020-03-24 | 江苏大学 | Method for improving ganoderma lucidum mycelium yield and triterpene yield by using low-intensity ultrasonic waves |
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| CN110903982A (en) * | 2019-11-14 | 2020-03-24 | 江苏大学 | Method for improving ganoderma lucidum mycelium yield and triterpene yield by using low-intensity ultrasonic waves |
| CN110903982B (en) * | 2019-11-14 | 2023-05-05 | 江苏大学 | Method for improving ganoderma lucidum mycelium yield and triterpene yield by using low-intensity ultrasonic waves |
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