CN106119313A - A kind of liquid fermentation method of phellinus igniarius mycelium - Google Patents
A kind of liquid fermentation method of phellinus igniarius mycelium Download PDFInfo
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Abstract
The invention provides the liquid fermentation method of a kind of phellinus igniarius mycelium, the method its be the four water magnesium acetates adding 0.7 1.1g/L in the liquid fermentation and culture of phellinus igniarius mycelium.The liquid fermentation method production technology of the phellinus igniarius mycelium of the present invention is simple, additive is with low cost, fermentation time is relatively short, and in the phellinus igniarius mycelium of generation, the content of flavone is high.
Description
Technical field
The invention belongs to technical field of biological fermentation, more particularly to the liquid fermentation method of a kind of phellinus igniarius mycelium.
Background technology
Phellinus igniarius (L. ex Fr.) Quel. (Sanghuangporus sanghuang), is commonly called as Phellinus igniarius (L. ex Fr.) Quel., Mulberry, records the earliest in Li Shizhen (1518-1593 A.D.) " book on Chinese herbal medicine guiding principle
Mesh " in, it is a kind of perennial medicinal fungi.The crucial active ingredient of Phellinus igniarius (L. ex Fr.) Quel. is phellin polysaccharides and flavonoids from phellinus, and wherein Phellinus igniarius (L. ex Fr.) Quel. is yellow
Ketone has the effects such as antitumor, antioxidation, enhancing immunity, is one of internationally recognized optimal anticancer epiphyte.
Present stage, the yield of Phellinus igniarius (L. ex Fr.) Quel. thalline and crucial active ingredient thereof becomes the bottleneck hindering Phellinus igniarius (L. ex Fr.) Quel. products application.Due to
Liquid submerged fermentation technology produce Phellinus igniarius (L. ex Fr.) Quel. there is the features such as quality controllable, with short production cycle, the most become acquisition Phellinus igniarius (L. ex Fr.) Quel. and
The effective ways of active substance.In recent years, the liquid fermentation technology of Phellinus igniarius (L. ex Fr.) Quel. has made great progress, but improves Mulberry the most further
The yield of yellow mycelium or crucial active ingredient, to improve its added value, is the Phellinus igniarius (L. ex Fr.) Quel. problem that develops that letter is to be solved.
Current study show that, coumarin, naphthalene acetic acid, cinnamic acid, sodium acetate can induce the synthesis of flavonoids from phellinus.These works
Good material and phenotype is provided as the research biosynthetic regulatory mechanism of flavonoids from phellinus.But above-mentioned inducer is to Phellinus igniarius (L. ex Fr.) Quel.
The effect of increasing production of flavone is the most relatively low, increases production flavonoids from phellinus the most in a large number and remains a need for probing into more effective inducer.
Summary of the invention
It is an object of the invention to provide the liquid fermentation method of a kind of phellinus igniarius mycelium, it is the liquid at phellinus igniarius mycelium
Body fermentation culture is added the four water magnesium acetates of 0.7-1.1g/L.
Wherein add four water magnesium acetates, can arbitrary sky in the 0th day, the 1st day, the 2nd day or the 3rd day in fermentation culture
Add.
Specifically, the liquid fermentation method of the phellinus igniarius mycelium of the present invention comprises the steps:
(1) Phellinus igniarius (L. ex Fr.) Quel. strain is inoculated in seed culture medium, after inoculation in 24-30 DEG C, cultivate 5-9 under 100-180r/min
My god, obtain seed culture fluid；
(2) by seed culture fluid, access in fermentation medium according to the inoculum concentration of 5-15% (relative to fermentation medium),
After inoculation in 24-30 DEG C, cultivate 6-10 days under 100-180r/min, and in incubation, add the four water second of 0.7-1.1g/L
Acid magnesium, cultivates and obtains phellinus igniarius mycelium.
Phellinus igniarius (L. ex Fr.) Quel. strain in wherein said step (1) is the inclined-plane mycelium Phellinus igniarius (L. ex Fr.) Quel. bacterium in slant medium after activation
Strain, wherein slant medium is: 39g potato dextrose agar is dissolved in 1L deionized water after sterilizing and get final product；
The component of the seed culture medium in described step (1) is: potato dextrose broth 24g/L, mulberry powder 10g/L,
PH is natural；
The component of the fermentation medium in described step (2) is: glucose 20g/L, yeast autolysis powder 10g/L, seven water sulfur
Acid magnesium 1g/L, potassium dihydrogen phosphate 1g/L, mulberry powder 10g/L, pH are natural；
Described step (2) is added in incubation the four water magnesium acetates of 0.7-1.1g/L, can be in fermentation culture
The 0th day, the 1st day, the 2nd day or the 3rd day in arbitrary day interpolation.
The mycelium of fermentation ends, is centrifuged 10min in 5000r/min, abandons supernatant, with distilled water wash centrifugation 3-
4 times, collection is deposited in 60 DEG C and is dried to constant weight.Measure the flavones content in mycelium, find to ferment far above conventional method
To mycelium in flavones content.
The present invention is directed to the problem that existing flavonoids from phellinus fermentation yield is the highest, it is provided that one can significantly improve phellinus liteus
The liquid fermentation method of flavones content in body, and production technology is simple, additive is with low cost, fermentation time is relatively short, raw
In the phellinus igniarius mycelium become, the content of flavone is high, can be applicable in the industrialized production of flavonoids from phellinus.
Detailed description of the invention
Raw material sources:
Phellinus igniarius (L. ex Fr.) Quel. bacterial strain be Phellinus igniarius (L. ex Fr.) Quel. Sanghuangporus sanghuang (this bacterial strain at fungus journal, 2016,35 (5):
1-12 delivers), by Edible Fungus Inst., Shanghai Academy of Agriculture's process technology and the preservation of fermentation engineering research department.
Potato dextrose agar and potato dextrose broth are purchased from U.S. company BD；
Embodiment 1
(1) prepared by seed culture medium: potato dextrose broth 24g/L, and mulberry powder 10g/L, pH are natural.
(2) by phellinus liteus inoculation 6-8 block good for slant culture, (every piece of area is about 0.03cm2-0.05cm2) in seed
In culture medium, in temperature 26 DEG C, rotating speed is to cultivate after 7 days under 150rpm, obtains seed culture fluid.
(3) by cultured seed culture fluid, the inoculum concentration according to 10% accesses fermentation medium (glucose 20g/L, ferment
Female self-dissolving powder 10g/L, Magnesium sulfate heptahydrate 1g/L, potassium dihydrogen phosphate 1g/L, mulberry powder 10g/L, pH are natural) in, in 26 after inoculation
DEG C, cultivate 6 days under 150rpm, within the 0th day, add four water magnesium acetates, interpolation concentration 0.9g/L of four water magnesium acetates in fermentation, collect
Mycelium after fermentation.
The mycelium collected is centrifuged 10min in 5000r/min, abandons supernatant, with distilled water wash centrifugation 3-4
Secondary, collection is deposited in 60 DEG C and is dried to constant weight.
Measuring flavonoids from phellinus content: pulverize last by dry phellinus igniarius mycelium, 69-69mg mycelium adds 2mL
Volume fraction is the ethanol of 80%, after extracting 2 hours in 40 DEG C ultrasonic (40KHz, 100W), extracting solution in 10000r/min from
Heart 8min, supernatant is testing sample solution.Add testing sample solution 1mL and (set three repetitions) in test tube, then it is sub-to add 5%
Sodium nitrate solution 1mL, after fully 6min is placed in mixing, then adds 10% 9 water aluminum nitrate solution 1mL, shakes up placement 6min；Add again
4% sodium hydroxide solution 10mL, afterwards with the ethanol constant volume that volume fraction is 80%, measures sample at 510nm after mixing
Absorbance, the flavones content of sample can be calculated according in rutin standard curve.The method is used to record the content of flavonoids from phellinus
For 7.48mg/100mg, compared with the 3.80mg/100mg of comparison (being not added with four water magnesium acetates), improve 96.84%.
Embodiment 2
(1) prepared by seed culture medium: potato dextrose broth 24g/L, and mulberry powder 10g/L, pH are natural..
(2) by phellinus liteus inoculation 6-8 block good for slant culture, (every piece of area is about 0.03cm2-0.05cm2) in seed
In culture medium, in temperature 26 DEG C, rotating speed is to cultivate after 7 days under 150rpm, obtains seed culture fluid.
(3) by cultured seed culture fluid, the inoculum concentration according to 10% accesses fermentation medium (glucose 20g/L, ferment
Female self-dissolving powder 10g/L, Magnesium sulfate heptahydrate 1g/L, potassium dihydrogen phosphate 1g/L, mulberry powder 10g/L, pH are natural) in, in 26 after inoculation
DEG C, cultivate 6 days under 150rpm, ferment the 1st day and add four water magnesium acetates, interpolation concentration 0.9g/L of four water magnesium acetates, collect and send out
Yeast-like fungi filament.
Collect the mycelium of fermentation ends in step (3), be centrifuged 10min in 5000r/min, abandon supernatant, use distilled water
Washing centrifugation 3-4 time, collection is deposited in 60 DEG C and is dried to constant weight.
Measure flavonoids from phellinus content: flavonoids from phellinus assay method is with embodiment 1.The method is used to record containing of flavonoids from phellinus
Amount is 8.86mg/100mg, compared with the 3.80mg/100mg of comparison (being not added with four water magnesium acetates), improves 133.16%.Implement
Example 3
(1) prepared by seed culture medium: potato dextrose broth 24g/L, and mulberry powder 10g/L, pH are natural.
(2) by phellinus liteus inoculation 6-8 block good for slant culture, (every piece of area is about 0.03cm2-0.05cm2) in seed
In culture medium, in temperature 26 DEG C, rotating speed is to cultivate after 7 days under 150rpm, obtains seed culture fluid.
(3) by cultured seed culture fluid, the inoculum concentration according to 10% accesses fermentation medium (glucose 20g/L, ferment
Female self-dissolving powder 10g/L, Magnesium sulfate heptahydrate 1g/L, potassium dihydrogen phosphate 1g/L, mulberry powder 10g/L, pH are natural) in, in 26 after inoculation
DEG C, to cultivate 6 days under 150rpm, ferment the 2nd day and add four water magnesium acetates, the interpolation concentration of four water magnesium acetates is 0.9g/L, collects
Mycelium after fermentation.
By the mycelium of fermentation ends, it is centrifuged 10min in 5000r/min, abandons supernatant, use distilled water wash centrifugation
3-4 time, collection is deposited in 60 DEG C and is dried to constant weight.
Measure flavonoids from phellinus content: flavonoids from phellinus assay method is with embodiment 1.The method is used to record containing of flavonoids from phellinus
Amount is 7.74mg/100mg, compared with the 3.80mg/100mg of comparison (being not added with four water magnesium acetates), improves 103.68%.Implement
Example 4
(1) prepared by seed culture medium: potato dextrose broth 24g/L, and mulberry powder 10g/L, pH are natural.
(2) by phellinus liteus inoculation 6-8 block good for slant culture, (every piece of area is about 0.03cm2-0.05cm2) in seed
In culture medium, in temperature 26 DEG C, rotating speed is to cultivate after 7 days under 150rpm, obtains seed culture fluid.
(3) by cultured seed culture fluid, the inoculum concentration according to 10% accesses fermentation medium (glucose 20g/L, ferment
Female self-dissolving powder 10g/L, Magnesium sulfate heptahydrate 1g/L, potassium dihydrogen phosphate 1g/L, mulberry powder 10g/L, pH are natural) in, in 26 after inoculation
DEG C, cultivate 6 days under 150rpm, ferment the 3rd day and add four water magnesium acetates, interpolation concentration 0.9g/L of four water magnesium acetates, collect and send out
Mycelium after ferment.
Mycelium after fermentation, is centrifuged 10min in 5000r/min, abandons supernatant, with distilled water wash centrifugation 3-4
Secondary, collection is deposited in 60 DEG C and is dried to constant weight.
Measure flavonoids from phellinus content: flavonoids from phellinus assay method is with embodiment 1.The method is used to record containing of flavonoids from phellinus
Amount is 6.12mg/100mg, compared with the 3.80mg/100mg of comparison (being not added with four water magnesium acetates), improves 61.05%.
Claims (4)
1. a liquid fermentation method for phellinus igniarius mycelium, is characterized in that adding in the liquid fermentation and culture of phellinus igniarius mycelium
The four water magnesium acetates of 0.7-1.1g/L.
The liquid fermentation method of phellinus igniarius mycelium the most according to claim 1, it is characterised in that wherein add 0.7-1.1g/
The four water magnesium acetates of L, are the interpolations in arbitrary day in the 0th day in fermentation culture, the 1st day, the 2nd day or the 3rd day.
The liquid fermentation method of phellinus igniarius mycelium the most according to claim 1, it is characterised in that comprise the steps:
(1) Phellinus igniarius (L. ex Fr.) Quel. strain is inoculated in seed culture medium, after inoculation in 24-30 DEG C, cultivate 5-9 days under 100-180r/min,
Obtain seed culture fluid；
(2) by seed culture fluid, access in fermentation medium according to the inoculum concentration of 5-15%, in 24-30 DEG C, 100-after inoculation
Cultivate 6-10 days under 180r/min, and in incubation, add the four water magnesium acetates of 0.7-1.1g/L, cultivate and obtain Phellinus igniarius (L. ex Fr.) Quel. bacterium
Filament.
The liquid fermentation method of phellinus igniarius mycelium the most according to claim 3, it is characterised in that wherein add 0.7-1.1g/
The four water magnesium acetates of L, are the interpolations in arbitrary day in the 0th day in fermentation culture, the 1st day, the 2nd day or the 3rd day.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112322572A (en) * | 2020-11-19 | 2021-02-05 | 陕西省微生物研究所 | Liquid fermentation method for increasing yield of phellinus igniarius mycelium |
| CN116240114A (en) * | 2023-01-31 | 2023-06-09 | 山东蓬勃生物科技有限公司 | Phellinus linteus YX2, extract and application thereof |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112322572A (en) * | 2020-11-19 | 2021-02-05 | 陕西省微生物研究所 | Liquid fermentation method for increasing yield of phellinus igniarius mycelium |
| CN116240114A (en) * | 2023-01-31 | 2023-06-09 | 山东蓬勃生物科技有限公司 | Phellinus linteus YX2, extract and application thereof |
| CN116240114B (en) * | 2023-01-31 | 2023-11-03 | 山东蓬勃生物科技有限公司 | Phellinus linteus YX2, extract and application thereof |
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