CN103392510B - Method for increasing mycelium ganodenic acid content of ganoderma lucidum liquid in fermentation - Google Patents
Method for increasing mycelium ganodenic acid content of ganoderma lucidum liquid in fermentation Download PDFInfo
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- CN103392510B CN103392510B CN201310348015.2A CN201310348015A CN103392510B CN 103392510 B CN103392510 B CN 103392510B CN 201310348015 A CN201310348015 A CN 201310348015A CN 103392510 B CN103392510 B CN 103392510B
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Abstract
The invention discloses a method for increasing the mycelium ganodenic acid content of ganoderma lucidum liquid in fermentation. Fermentation of the ganoderma lucidum liquid is divided into two stages, wherein the first-stage fermentation facilitates fast growth of ganoderma lucidum hyphae, and the second-stage fermentation induces the mycelium of the ganoderma lucidum to increase the ganodenic acid content by adding acetic acid into fermentation liquid obtained by the first-stage fermentation. The method for increasing the mycelium ganodenic acid content of ganoderma lucidum liquid in fermentation can obviously increase the ganodenic acid content of the ganoderma lucidum mycelium liquid in fermentation, has the advantages of being safe in food grade, low in cost, free of pollution, simple in method, capable of easily achieving industrial production and the like, and therefore lays the foundation of industrial economical production for the ganodenic acid.
Description
Technical field
The present invention relates to a kind of method utilizing acetic acid to carry out two benches fermentation raising liquid fermentation mycelium of lucid ganoderma ganoderic acid content.Be specifically related to a kind of in ganoderma lucidum mycelium liquid fermentation, the ramp of first stage promotion Ganoderma lucidum mycelium, subordinate phase induces Ganoderma mycelium to improve ganoderic acid content by adding acetic acid.
Background technology
Glossy ganoderma (Ganoderma lucidum) is Basidiomycetes, polyporaceae, glossy ganoderma Pseudomonas fungi, is the foremost high medicinal fungi of China.The history of more than 2000 year is had as medicine.In glossy ganoderma, study more chemical composition has the materials such as polyose, triterpenes, ucleosides, sterols, alkaloids, furans, amino acid polypeptide class, lipid, trace element at present.Ganodenic acid is one of main active ingredient of glossy ganoderma, meanwhile, is also primary medicinal component.According to the literature, Ganodenic acid has antitumor, hepatoprotective effect, suppresses histamine release, angiotensin-converting enzyme isoreactivity.And impel by various method (as ultraviolet mutagenesis seed selection) the effective active composition obtaining more glossy ganoderma, the focus having become Chinese scholars research and paid close attention to.Improve ganoderic acid content, have great meaning to the medicinal effect improving glossy ganoderma.
Current research shows, methyl jasmonate, phenylethyl barbituric acid, acetylsalicylic acid and cupric ion can both induce Ganodenic acid to synthesize.These work provides good material and phenotype for studying the biosynthetic regulatory mechanism of Ganodenic acid.But Ganodenic acid leavened prod is as food and medicine, its security of fermenting and economy must be considered.Methyl jasmonate itself is expensive, and phenylethyl barbituric acid is a kind of ubiquity central nervous depressant, and acetylsalicylic acid is a kind of antipyretic and analgesic, and cupric ion is a heavy metal species.In Ganodenic acid fermentation actual production, need one have aliment security level other, cheap inductor, and acetic acid is the main component of edible vinegar, has edible safety and price is lower.
Summary of the invention
The object of the present invention is to provide a kind of method improving liquid fermentation mycelium of lucid ganoderma ganoderic acid content.
Object of the present invention can be achieved through the following technical solutions:
A kind of method improving liquid fermentation mycelium of lucid ganoderma ganoderic acid content, this Ganoderma lucidum submerged fermentation is divided into two stage fermentations: first stage fermentation promotes Ganoderma lucidum mycelium ramp, and subordinate phase ferment and added acetic acid in the fermented liquid that obtains by fermenting to the first stage and induce Ganoderma mycelium raising ganoderic acid content.
Above-mentioned method, comprises the following steps:
(1) to CYM inoculation of medium Liquid Strain of Ganoderma Lucidum, temperature 28 ~ 30 DEG C, under rotating speed 120 ~ 150 r/min condition, carry out first stage fermentation, fermentation time is 120 ~ 168 hours;
(2) after first stage fermentation ends, in fermented liquid, add acetic acid, proceed subordinate phase fermentation, fermentation time is 8 ~ 48 hours.
The interpolation concentration of described acetic acid is 2 mM ~ 14mM; Be preferably 5 mM ~ 14mM.
The inoculum size of described liquid spawn is 3% ~ 15% of CYM culture volume.
The formula of 1 liter of described CYM substratum is: maltose 10 grams, glucose 20 grams, yeast extract 2 grams, peptone 2 grams, magnesium sulfate heptahydrate 0.5 gram, and potassium primary phosphate 1 gram, is settled to 1 liter with water, 115 DEG C of sterilizings 25 ~ 40 minutes.
The method of raising liquid fermentation mycelium of lucid ganoderma ganoderic acid content of the present invention, adds acetic acid and induces glossy ganoderma to produce more Ganodenic acid when ganoderma lucidum mycelium liquid ferments.The inductor acetic acid safety used, and cheap, the output of Ganodenic acid can be significantly improved.
Beneficial effect of the present invention
By method of the present invention, ganoderic acid content when ganoderma lucidum mycelium liquid ferments can be significantly improved, and it is safe and reliable, method is simple, with low cost, adopt the inventive method that the content of Ganodenic acid in the Ganoderma mycelium of liquid fermenting can be made to reach 2.77 ~ 5.74 mg/100 mg DW compared with conventional culture methods, relative to comparative example ganoderic acid content 2.17 mg/100 mg DW, improve 27.6 % ~ 156.2 %.
Embodiment
Below in conjunction with example, the present invention is described in further details:
Embodiment one
1. configure CYM substratum, filling a prescription is: maltose 10 grams, glucose 20 grams, yeast extract 2 grams, peptone 2 grams, magnesium sulfate heptahydrate 0.5 gram, and potassium primary phosphate 1 gram, is settled to 1 liter with water, and sterilising conditions is 115 DEG C of sterilizings 30 minutes.
2. in the CYM substratum configured, access Liquid Strain of Ganoderma Lucidum with the inoculum size of 5 %, temperature 28 DEG C, under rotating speed 150 r/min condition, carry out the fermentation of first stage, fermentation time is respectively 128, and 136,144,152,160,168 hours.
3., respectively after first stage fermentation ends, aseptically respectively to adding the acetic acid that ultimate density is 5 mM in fermented liquid, carry out subordinate phase fermentation.
4. after subordinate phase fermentation ends, collect mycelium pellet with copper filter screen, and with distilled water flushing 5 ~ 6 times, dry in 60 DEG C of baking ovens to constant weight, spectrophotometry ganoderic acid content.Get hypha powder 0.50 about the g of oven dry, be placed in 25 mL volumetric flasks and add 95% ethanol constant volume, in sonicator, ultrasonication is after 2 hours, get centrifugal 10 min of supersound extraction liquid 4000 r/min, get supernatant liquor 0.10 mL in 10 mL tool plug test tubes, add Vanillin 0.20 mL successively, perchloric acid 0.50 mL, mixing, 60 DEG C of water-bath 20 min, take out and cool immediately, add 5.00 mL Glacial acetic acid, measure absorbancy in 550 nm places after mixing, measure ganoderic acid content.Detected result is in table 1.Content increases obviously compared with comparative example, and ganoderic acid content increases per-cent in table 1.
Embodiment two
1. configure CYM substratum, filling a prescription is: maltose 10 grams, glucose 20 grams, yeast extract 2 grams, peptone 2 grams, magnesium sulfate heptahydrate 0.5 gram, and potassium primary phosphate 1 gram, is settled to 1 liter with water, and sterilising conditions is 115 DEG C of sterilizings 30 minutes.
2. in the CYM substratum configured, access Liquid Strain of Ganoderma Lucidum with the inoculum size of 5 %, temperature 28 DEG C, under rotating speed 150 r/min condition, carry out the fermentation of first stage, fermentation time is 120 hours.
3. when first stage fermentation ends, aseptically respectively to the acetic acid adding different concns in fermented liquid, carry out subordinate phase fermentation, fermentation time is 48 hours.The ultimate density of adding acetic acid in secondary fermentation liquid is respectively 2 mM, 4 mM, 6 mM, 8 mM, 10 mM, 12 mM, 14 mM.
4. after subordinate phase fermentation ends, collect mycelium pellet with copper filter screen, and with distilled water flushing 5 ~ 6 times, dry in 60 DEG C of baking ovens to constant weight, spectrophotometry ganoderic acid content.Get hypha powder 0.50 about the g of oven dry, be placed in 25 mL volumetric flasks and add 95% ethanol constant volume, in sonicator, ultrasonication is after 2 hours, get centrifugal 10 min of supersound extraction liquid 4000 r/min, get supernatant liquor 0.10 mL in 10 mL tool plug test tubes, add Vanillin 0.20 mL successively, perchloric acid 0.50 mL, mixing, 60 DEG C of water-bath 20 min, take out and cool immediately, add 5.00 mL Glacial acetic acid, measure absorbancy in 550 nm places after mixing, measure ganoderic acid content.Detected result is in table 1.Content increases obviously compared with comparative example, and ganoderic acid content increases per-cent in table 2.
Comparative example
1. according to the method preparation CYM substratum in embodiment 1, if three repetitions.115 DEG C of sterilizings 30 minutes.With 5 % inoculum size access Liquid Strain of Ganoderma Lucidums, 28 DEG C, 150 r/min cultivate 168 hours.
2. after fermentation ends, collect mycelium pellet with copper filter screen, and with distilled water flushing 5 ~ 6 times, dry in 60 DEG C of baking ovens to constant weight, spectrophotometry ganoderic acid content.Get hypha powder 0.50 about the g of oven dry, be placed in 25 mL volumetric flasks and add 95% ethanol constant volume, in sonicator, ultrasonication is after 2 hours, get centrifugal 10 min of supersound extraction liquid 4000 r/min, get supernatant liquor 0.10 mL in 10 mL tool plug test tubes, add Vanillin 0.20 mL successively, perchloric acid 0.50 mL, mixing, 60 DEG C of water-bath 20 min, take out and cool immediately, add 5.00 mL Glacial acetic acid, measure absorbancy in 550 nm places after mixing, measure ganoderic acid content.Recording ganoderic acid content is 2.17mg/100mg.
The culture medium culturing ganoderic acid content of table 1 first stage incubation time and the different induction time of 5 mM acetic acid
The culture medium culturing ganoderic acid content of table 2 subordinate phase different concns acetic acid
| Subordinate phase acetic acid concentration mM | Ganoderic acid content mg/100 mg | Ganoderic acid content improves per-cent % |
| 0 | 2.17 | - |
| 2 | 2.77 | 27.6 |
| 4 | 3.02 | 39.2 |
| 6 | 4.73 | 118.0 |
| 8 | 5.32 | 145.2 |
| 10 | 5.56 | 156.2 |
| 12 | 5.17 | 138.2 |
| 14 | 4.32 | 99.1 |
Claims (4)
1. one kind is improved the method for liquid fermentation mycelium of lucid ganoderma ganoderic acid content, it is characterized in that this Ganoderma lucidum submerged fermentation is divided into two stage fermentations: first stage fermentation promotes Ganoderma lucidum mycelium ramp, subordinate phase ferment and is added acetic acid in the fermented liquid that obtains by fermenting to the first stage and induce Ganoderma mycelium raising ganoderic acid content;
Comprise the following steps:
(1) to CYM inoculation of medium Liquid Strain of Ganoderma Lucidum, temperature 28 ~ 30 DEG C, under rotating speed 120 ~ 150r/min condition, carry out first stage fermentation, fermentation time is 120 ~ 168 hours;
(2) after first stage fermentation ends, in fermented liquid, add acetic acid, proceed subordinate phase fermentation, fermentation time is 8 ~ 48 hours;
The interpolation concentration of described acetic acid is 2mM ~ 14mM.
2. method according to claim 1, is characterized in that the interpolation concentration of described acetic acid is 5mM ~ 14mM.
3. method according to claim 1, is characterized in that the inoculum size of described liquid spawn is 3% ~ 15% of CYM culture volume.
4. method according to claim 1, is characterized in that the formula of 1 liter of described CYM substratum is: maltose 10 grams, glucose 20 grams, yeast extract 2 grams, peptone 2 grams, magnesium sulfate heptahydrate 0.5 gram, potassium primary phosphate 1 gram, is settled to 1 liter with water, 115 DEG C of sterilizings 25 ~ 40 minutes.
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| CN107603987B (en) * | 2017-10-16 | 2020-06-09 | 南京农业大学 | Application of ganoderma lucidum transcription factor SKN7 gene in regulation and control of ganoderma lucidum triterpene biosynthesis |
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