CN101353676A - Ganoderma lucidum mycelium liquid fermentation method improving ganoderic acid content with fungal elicitor - Google Patents
Ganoderma lucidum mycelium liquid fermentation method improving ganoderic acid content with fungal elicitor Download PDFInfo
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Abstract
The invention relates to a Ganoderma lucidum mycelium liquid fermentation method for improving ganoderic acid content by utilizing Fungal Elicitors. The method comprises the steps of the preparation of the Fungal Elicitors, the liquid fermentation culture of the Ganoderma lucidum and the co-culture of the Ganoderma lucidum and the Fungal Elicitors. The method has the advantages of simple process, low cost, obvious effect, high content of the ganoderic acid, and being a manufacturing method with huge industrial prospect.
Description
Technical field:
The invention belongs to technical field of bioengineering, be specifically related to a kind of liquid fermentation process that utilizes fungal elicitor production high yield Ganodenic acid.
Background technology:
Glossy ganoderma (Ganoderma lucidum) is Basidiomycetes, polyporaceae, glossy ganoderma Pseudomonas fungi.Modern pharmacology studies show that glossy ganoderma have antitumor, anti-ageing, prevent effects such as arteriosclerosis, and cardiovascular and cerebrovascular diseases is had higher pharmacologically active.Compositions such as the polysaccharide in the glossy ganoderma, triterpene substance (being mainly Ganodenic acid), alkaloid, amino acid polypeptide, trace element are the effective substances of glossy ganoderma.Wherein Ganodenic acid is glossy ganoderma treatment hypertension, hyperlipidemia, insomnia and various respiratory systemic disease (as go phlegm, cough-relieving, relieving asthma) and antitumor etc. basic substance.When the glossy ganoderma pharmaceutical use was extensively disclosed, the output of glossy ganoderma became the limiting factor that suppresses the glossy ganoderma application, because very rare at the occurring in nature wild Ganoderma, and wherein ganoderic acid content only is 1 milligram/100 milligrams dry weights, far can not satisfy people's needs.Compare with the glossy ganoderma artificial culture, the fermentative Production effective component of glossy ganoderma since with short production cycle, labor force economize and be subjected to advantages such as external environment influence is little to be considered to a kind of effective means.
In recent years, the extract that utilizes specific fungal bacterial strain improves the accumulation volume of some secondary metabolite in the vegetable cell as elicitor, has obtained immense success in the vegetable cell suspension culture.But, the report that fungal elicitor is used for the edible and medicinal fungi cell cultures seldom, at present rarely seen about improving the report that the cordycepin liquid fermenting is produced with fungal elicitor, do not appear in the newspapers as yet with the research that improves secondary meta-bolitess such as Ganodenic acid output and be used for the glossy ganoderma fermentation cultivation.
Summary of the invention:
The objective of the invention is to overcome the deficiency of above-mentioned prior art and provide that a kind of technology is simple, cost is low, effect is remarkable, the liquid fermentation process that utilizes fungal elicitor production high yield Ganodenic acid that ganoderic acid content is high, present method can be used for the suitability for industrialized production of Ganodenic acid.
Purpose of the present invention can reach by following measure: a kind of ganoderma lucidum mycelium liquid fermentation method that utilizes fungal elicitor to improve ganoderic acid content is characterized in that comprising the steps:
(1) preparation of fungal elicitor:
1. the cultivation of fungal elicitor bacterial strain: with the fungal elicitor bacterial classification inoculation on the PDA slant medium activation after, be transferred on the PDA plate culture medium, cultivated 3～5 days for 25～30 ℃, getting the bacterium piece is inoculated in the triangular flask that the PDA liquid nutrient medium is housed, after in 25～30 ℃, 100～180r/min shaking table, cultivating 5～7 days, filter that to obtain mycelium standby;
2. fungal elicitor preparation: the fungal elicitor mycelium is dried down in 60～70 ℃, grind broken, 1: 2～1: 5 ratio of volume ratio adding concentration is 65～85% ethanol by weight, at room temperature soaked 10～15 hours, decompress filter, collect filter residue, filter residue 1: 2～1: 5 ratio of volume ratio by weight adds the mixed solution of chloroform and propyl carbinol, the volume ratio of chloroform and propyl carbinol is 3: 1～5: 1, filter wash slag 3～5 times, then filter residue by weight 1: 3～1: 5 ratio of volume ratio with acetone rinsing after, filter residue 1: 3～1: 5 ratio of volume ratio water flushing more by weight, natural air drying, dry-matter 1: 5～1: 7 ratio of volume ratio adding concentration by weight is 5～15% trifluoroacetic acids, boiling water bath acidolysis 0.5～2 hour, and hydrolyzate filters, get filtrate, being neutralized to pH with NaOH is 4～7, leaves standstill 10～15 hours filtration sterilization under 3 ℃～8 ℃, make fungal elicitor, freezing preservation;
(2) liquid fermentation and culture of glossy ganoderma:
The Ganderma lucidum strain of slant preservation is inoculated on the PDA plate culture medium, carry out activation culture, 25～30 ℃ of culture temperature, incubation time 5～10 days, getting the bacterium piece then is inoculated in the intermediate cam bottle that liquid fermentation medium is housed and carries out fermentation culture, shaking speed 100～180r/min, 25～30 ℃ of temperature, 5～8 days time;
Liquid fermentation medium is formed: glucose 20～50g/L, yeast extract paste 2～8g/L, peptone 2～8g/L, potassium primary phosphate 0.5～2.0g/L, sal epsom 0.1～1.0g/L, vitamins B
10.01～0.10g/L, surplus is a distilled water;
(3) glossy ganoderma and fungal elicitor are cultivated altogether:
Ganoderma lucidum liquid fermentation culture the 1st～5 day, add fungal elicitor, make fungal elicitor concentration reach 40～120 μ g/mL, continue then to cultivate 2～7 days, obtain Ganoderma mycelium.
In order further to realize purpose of the present invention, described fungal elicitor bacterial classification is Acremonium fungi top spore (Acremonium strictum), bacterium numbering CGMCC No.3.2058; Or fungus Trichoderma lignin wood mould (Trichoderma viride), bacterium numbering is CGMCC No.3.2196; Or Verticillium genus fungi mushroom wheel branch spore (Verticillium psalliotae), bacterium numbering is CGMCC No.3.4423.
In order further to realize purpose of the present invention, described fermentation Ganderma lucidum strain is common cultivation kind (Ganoderma lucidum), available from Chinese common micro-organisms DSMZ, is numbered CGMCC No.5.644.
In order further to realize purpose of the present invention, 3～5 of the bacterium pieces of cut-off footpath 0.3～0.6cm are inoculated in the PDA liquid nutrient medium the bottled PDA liquid nutrient medium of every 500mL triangle 100～200mL in the culturing step of described fungal elicitor bacterial strain.
In order further to realize purpose of the present invention, 3～5 of the bacterium pieces of cut-off footpath 0.3～0.6cm are inoculated in the liquid fermentation medium the bottled PDA liquid nutrient medium of every 500mL triangle 100～200mL in the liquid fermentation and culture step of described glossy ganoderma.
In order further to realize purpose of the present invention, fungal elicitor adding concentration is 80 μ g/mL in the common culturing step of described glossy ganoderma and fungal elicitor.
In order to realize that further purpose of the present invention, described glossy ganoderma and fungal elicitor are total to the adding in the 1st day that fungal elicitor is cultivated in glossy ganoderma fermentation in the culturing step, then with the glossy ganoderma co-cultivation.
In order to realize that further purpose of the present invention, described glossy ganoderma and fungal elicitor are total to the adding in the 3rd day that fungal elicitor is cultivated in glossy ganoderma fermentation in the culturing step, then with the glossy ganoderma co-cultivation.
In order to realize that further purpose of the present invention, described glossy ganoderma and fungal elicitor are total to the adding in the 5th day that fungal elicitor is cultivated in glossy ganoderma fermentation in the culturing step, then with the glossy ganoderma co-cultivation.
The present invention can produce following positively effect compared with the prior art: the present invention is used for fungal elicitor the liquid fermenting production of Ganodenic acid, increase substantially the content (bringing up to the 3.2mg/100mg mycelia) of Ganodenic acid by the 0.8mg/100mg mycelia, this novel method operating procedure is simple, with low cost, have good industrial applications prospect.
Embodiment:
Below the specific embodiment of the present invention is elaborated:
Embodiment 1:
Bacterial classification: the fungal elicitor bacterial classification is Acremonium fungi top spore (Acremoniumstrictum), is bought by Chinese common micro-organisms DSMZ, and bacterium numbering is CGMCCNo.3.2058.
Ganderma lucidum strain is common cultivation kind (Ganoderma lucidum), available from Chinese common micro-organisms DSMZ, is numbered CGMCC No.5.644.
A kind of ganoderma lucidum mycelium liquid fermentation method that utilizes fungal elicitor to improve ganoderic acid content is characterized in that comprising the steps:
(1) preparation of fungal elicitor:
1. the cultivation of fungal elicitor bacterial strain: with the fungal elicitor bacterial classification inoculation on the PDA slant medium activation after, be transferred on the PDA plate culture medium, cultivated 5 days for 25 ℃, beat 3 of the bacterium pieces of cut-off footpath 0.3cm at colony edge with punch tool, be inoculated in the PDA liquid nutrient medium, the bottled PDA liquid nutrient medium of every 500mL triangle 100mL cultivated in 25 ℃, 100r/min shaking table after 7 days, filtered that to obtain mycelium standby;
2. fungal elicitor preparation: the fungal elicitor mycelium is dried down in 60 ℃, grind broken, 1: 2 ratio of volume ratio adding concentration is 65% ethanol by weight, at room temperature soaked 10 hours, decompress filter is collected filter residue, and filter residue 1: 2 ratio of volume ratio by weight adds the mixed solution of chloroform and propyl carbinol, the volume ratio of chloroform and propyl carbinol is 3: 1, filter wash slag 3～5 times, then filter residue by weight 1: 3 ratio of volume ratio with acetone rinsing after, filter residue 1: 3 ratio of volume ratio water flushing more by weight, natural air drying, dry-matter 1: 5 ratio of volume ratio adding concentration by weight is 5% trifluoroacetic acid, boiling water bath acidolysis 0.5 hour, and hydrolyzate filters, get filtrate, being neutralized to pH with NaOH is 5.8, leaves standstill filtration sterilization under 3 ℃ 10 hours, make fungal elicitor, freezing preservation;
(2) liquid fermentation and culture of glossy ganoderma:
The Ganderma lucidum strain of slant preservation is inoculated on the PDA plate culture medium, carry out activation culture, 25 ℃ of culture temperature, incubation time 10 days is beaten 3 of the bacterium pieces of cut-off footpath 0.3cm then at colony edge with punch tool, be inoculated in and carry out fermentation culture in the liquid fermentation medium, the bottled substratum 100mL of every 500mL triangle,, shaking speed 100r/min, 25 ℃ of temperature, 8 days time;
Liquid fermentation medium is formed: glucose 20g/L, yeast extract paste 2g/L, peptone 2g/L, potassium primary phosphate 0.5g/L, sal epsom 0.1g/L, vitamins B
10.01g/L surplus is a distilled water;
(3) glossy ganoderma and fungal elicitor are cultivated altogether:
Ganoderma lucidum liquid fermentation culture the 1st day, add fungal elicitor, make the fungal elicitor final concentration reach 40 μ g/mL, continue then to cultivate 7 days, obtain Ganoderma mycelium.
(4) extraction of Ganodenic acid and detection:
With the Ganoderma mycelium oven dry, get 0.5g sample (dry weight) grinding powder, add 30mL methyl alcohol supersound extraction 2 hours, filter, the supernatant evaporate to dryness adds methanol constant volume to 2mL, and ultraviolet colorimetric method for determining ganoderic acid content is 3.2mg/100mg mycelia (dry weight).
Embodiment 2:
Bacterial classification: the fungal elicitor bacterial classification is fungus Trichoderma lignin wood mould (Trichodermaviride), is bought by Chinese common micro-organisms DSMZ, and bacterium numbering is CGMCCNo.3.2196.
Ganderma lucidum strain is common cultivation kind (Ganoderma lucidum), available from Chinese common micro-organisms DSMZ, is numbered CGMCC No.5.644.
A kind of ganoderma lucidum mycelium liquid fermentation method that utilizes fungal elicitor to improve ganoderic acid content is characterized in that comprising the steps:
(1) preparation of fungal elicitor:
1. the cultivation of fungal elicitor bacterial strain: with the fungal elicitor bacterial classification inoculation on the PDA slant medium activation after, be transferred on the PDA plate culture medium, cultivated 3 days for 30 ℃, beat 5 of the bacterium pieces of cut-off footpath 0.6cm at colony edge with punch tool, be inoculated in the PDA liquid nutrient medium, the bottled PDA liquid nutrient medium of every 500mL triangle 200mL cultivated in 30 ℃, 180r/min shaking table after 5 days, filtered that to obtain mycelium standby;
2. fungal elicitor preparation: the fungal elicitor mycelium is dried down in 70 ℃, grind broken, 1: 5 ratio of volume ratio adding concentration is 85% ethanol by weight, at room temperature soaked 15 hours, decompress filter is collected filter residue, and filter residue 1: 5 ratio of volume ratio by weight adds the mixed solution of chloroform and propyl carbinol, the volume ratio of chloroform and propyl carbinol is 5: 1, filter wash slag 5 times, then filter residue by weight 1: 5 ratio of volume ratio with acetone rinsing after, filter residue 1: 5 ratio of volume ratio water flushing more by weight, natural air drying, dry-matter 1: 7 ratio of volume ratio adding concentration by weight is 15% trifluoroacetic acid, boiling water bath acidolysis 2 hours, and hydrolyzate filters, get filtrate, being neutralized to pH with NaOH is 4, leaves standstill filtration sterilization under 8 ℃ 15 hours, make fungal elicitor, freezing preservation;
(2) liquid fermentation and culture of glossy ganoderma:
The Ganderma lucidum strain of slant preservation is inoculated on the PDA plate culture medium, carry out activation culture, 30 ℃ of culture temperature, incubation time 5 days is beaten 5 of the bacterium pieces of cut-off footpath 0.6cm then at colony edge with punch tool, be inoculated in and carry out fermentation culture in the liquid fermentation medium, the bottled substratum 200mL of every 500mL triangle,, shaking speed 180r/min, 30 ℃ of temperature, 5 days time;
Liquid fermentation medium is formed: glucose 50g/L, yeast extract paste 8g/L, peptone 8g/L, potassium primary phosphate 2.0g/L, sal epsom 1.0g/L, vitamins B
10.10g/L surplus is a distilled water;
(3) glossy ganoderma and fungal elicitor are cultivated altogether:
Ganoderma lucidum liquid fermentation culture the 5th day, add fungal elicitor, make the fungal elicitor final concentration reach 120 μ g/mL, continue then to cultivate 2 days, obtain Ganoderma mycelium.
(4) extraction of Ganodenic acid and detection:
With the Ganoderma mycelium oven dry, get 1g sample (dry weight) grinding powder, add 50mL methyl alcohol supersound extraction 5 hours, filter, the supernatant evaporate to dryness adds methanol constant volume to 5mL, and ultraviolet colorimetric method for determining ganoderic acid content is 2.5mg/100mg mycelia (dry weight).
Embodiment 3:
Bacterial classification: the fungal elicitor bacterial classification is that Verticillium belongs to fungi mushroom wheel branch spore (Verticilliumpsalliotae), is bought by Chinese common micro-organisms DSMZ, and bacterium numbering is CGMCC No.3.4423.
Ganderma lucidum strain is common cultivation kind (Ganoderma lucidum), available from Chinese common micro-organisms DSMZ, is numbered CGMCC No.5.644.
A kind of ganoderma lucidum mycelium liquid fermentation method that utilizes fungal elicitor to improve ganoderic acid content is characterized in that comprising the steps:
(1) preparation of fungal elicitor:
1. the cultivation of fungal elicitor bacterial strain: with the fungal elicitor bacterial classification inoculation on the PDA slant medium activation after, be transferred on the PDA plate culture medium, cultivated 4 days for 28 ℃, beat 4 of the bacterium pieces of cut-off footpath 0.4cm at colony edge with punch tool, be inoculated in the PDA liquid nutrient medium, the bottled PDA liquid nutrient medium of every 500mL triangle 150mL cultivated in 28 ℃, 150r/min shaking table after 6 days, filtered that to obtain mycelium standby;
2. fungal elicitor preparation: the fungal elicitor mycelium is dried down in 65 ℃, grind broken, 1: 3 ratio of volume ratio adding concentration is 75% ethanol by weight, at room temperature soaked 12 hours, decompress filter is collected filter residue, and filter residue 1: 4 ratio of volume ratio by weight adds the mixed solution of chloroform and propyl carbinol, the volume ratio of chloroform and propyl carbinol is 4: 1, filter wash slag 4 times, then filter residue by weight 1: 4 ratio of volume ratio with acetone rinsing after, filter residue 1: 4 ratio of volume ratio water flushing more by weight, natural air drying, dry-matter 1: 6 ratio of volume ratio adding concentration by weight is 10% trifluoroacetic acid, boiling water bath acidolysis 1.5 hours, and hydrolyzate filters, get filtrate, being neutralized to pH with NaOH is 7, leaves standstill filtration sterilization under 5 ℃ 12 hours, make fungal elicitor, freezing preservation;
(2) liquid fermentation and culture of glossy ganoderma:
The Ganderma lucidum strain of slant preservation is inoculated on the PDA plate culture medium, carry out activation culture, 28 ℃ of culture temperature, incubation time 8 days is beaten 4 of the bacterium pieces of cut-off footpath 0.4cm then at colony edge with punch tool, be inoculated in and carry out fermentation culture in the liquid fermentation medium, the bottled substratum 150mL of every 500mL triangle,, shaking speed 150r/min, 28 ℃ of temperature, 7 days time;
Liquid fermentation medium is formed: glucose 35g/L, yeast extract paste 5g/L, peptone 5g/L, potassium primary phosphate 1g/L, sal epsom 0.5g/L, vitamins B
10.05g/L surplus is a distilled water;
(3) glossy ganoderma and fungal elicitor are cultivated altogether:
Ganoderma lucidum liquid fermentation culture the 3rd day, add fungal elicitor, make the fungal elicitor final concentration reach 80 μ g/mL, continue then to cultivate 5 days, obtain Ganoderma mycelium.
(4) extraction of Ganodenic acid and detection:
With the Ganoderma mycelium oven dry, get 0.8g sample (dry weight) grinding powder, add 40mL methyl alcohol supersound extraction 3 hours, filter, the supernatant evaporate to dryness adds methanol constant volume to 4mL, and ultraviolet colorimetric method for determining ganoderic acid content is 2.3mg/100mg mycelia (dry weight).
Fungal elicitor bacterial classification of the present invention also can adopt other similar bacterial classifications, and Ganderma lucidum strain also can adopt other common cultivation kinds.
Claims (9)
1, a kind of ganoderma lucidum mycelium liquid fermentation method that utilizes fungal elicitor to improve ganoderic acid content is characterized in that comprising the steps:
(1) preparation of fungal elicitor:
1. the cultivation of fungal elicitor bacterial strain: with the fungal elicitor bacterial classification inoculation on the PDA slant medium activation after, be transferred on the PDA plate culture medium, cultivated 3～5 days for 25～30 ℃, getting the bacterium piece is inoculated in the triangular flask that the PDA liquid nutrient medium is housed, after in 25～30 ℃, 100～180r/min shaking table, cultivating 5～7 days, filter that to obtain mycelium standby;
2. fungal elicitor preparation: the fungal elicitor mycelium is dried down in 60～70 ℃, grind broken, 1: 2～1: 5 ratio of volume ratio adding concentration is 65～85% ethanol by weight, at room temperature soaked 10～15 hours, decompress filter, collect filter residue, filter residue 1: 2～1: 5 ratio of volume ratio by weight adds the mixed solution of chloroform and propyl carbinol, the volume ratio of chloroform and propyl carbinol is 3: 1～5: 1, filter wash slag 3～5 times, then filter residue by weight 1: 3～1: 5 ratio of volume ratio with acetone rinsing after, filter residue 1: 3～1: 5 ratio of volume ratio water flushing more by weight, natural air drying, dry-matter 1: 5～1: 7 ratio of volume ratio adding concentration by weight is 5～15% trifluoroacetic acids, boiling water bath acidolysis 0.5～2 hour, and hydrolyzate filters, get filtrate, being neutralized to pH with NaOH is 4～7, leaves standstill 10～15 hours filtration sterilization under 3 ℃～8 ℃, make fungal elicitor, freezing preservation;
(2) liquid fermentation and culture of glossy ganoderma:
The Ganderma lucidum strain of slant preservation is inoculated on the PDA plate culture medium, carry out activation culture, 25～30 ℃ of culture temperature, incubation time 5～10 days, getting the bacterium piece then is inoculated in the intermediate cam bottle that liquid fermentation medium is housed and carries out fermentation culture, shaking speed 100～180r/min, 25～30 ℃ of temperature, 5～8 days time;
Liquid fermentation medium is formed: glucose 20～50g/L, yeast extract paste 2～8g/L, peptone 2～8g/L, potassium primary phosphate 0.5～2.0g/L, sal epsom 0.1～1.0g/L, vitaminB10 .01～0.10g/L, and surplus is a distilled water;
(3) glossy ganoderma and fungal elicitor are cultivated altogether:
Ganoderma lucidum liquid fermentation culture the 1st～5 day, add fungal elicitor, make fungal elicitor concentration reach 40～120 μ g/mL, continue then to cultivate 2～7 days, obtain Ganoderma mycelium.
2, a kind of ganoderma lucidum mycelium liquid fermentation method that utilizes fungal elicitor to improve ganoderic acid content according to claim 1, it is characterized in that described fungal elicitor bacterial classification is Acremonium fungi top spore (Acremonium strictum), bacterium numbering CGMCCNo.3.2058; Or fungus Trichoderma lignin wood mould (Trichoderma viride), bacterium numbering is CGMCC No.3.2196; Or Verticillium genus fungi mushroom wheel branch spore (Verticilliumpsalliotae), bacterium numbering is CGMCC No.3.4423.
3, a kind of ganoderma lucidum mycelium liquid fermentation method that utilizes fungal elicitor to improve ganoderic acid content according to claim 1, it is characterized in that described fermentation Ganderma lucidum strain is common cultivation kind (Ganoderma lucidum), available from Chinese common micro-organisms DSMZ, be numbered CGMCC No.5.644.
4, a kind of ganoderma lucidum mycelium liquid fermentation method that utilizes fungal elicitor to improve ganoderic acid content according to claim 1,3～5 of bacterium pieces that it is characterized in that cut-off footpath 0.3～0.6cm in the culturing step of described fungal elicitor bacterial strain, be inoculated in the PDA liquid nutrient medium the bottled PDA liquid nutrient medium of every 500mL triangle 100～200mL.
5, a kind of ganoderma lucidum mycelium liquid fermentation method that utilizes fungal elicitor to improve ganoderic acid content according to claim 1,3～5 of bacterium pieces that it is characterized in that cut-off footpath 0.3～0.6cm in the liquid fermentation and culture step of described glossy ganoderma, be inoculated in the liquid fermentation medium the bottled PDA liquid nutrient medium of every 500mL triangle 100～200mL.
6, a kind of ganoderma lucidum mycelium liquid fermentation method that utilizes fungal elicitor to improve ganoderic acid content according to claim 1 is characterized in that fungal elicitor adding concentration is 80 μ g/mL in the common culturing step of described glossy ganoderma and fungal elicitor.
7, a kind of ganoderma lucidum mycelium liquid fermentation method that utilizes fungal elicitor to improve ganoderic acid content according to claim 1, it is characterized in that described glossy ganoderma and fungal elicitor are total to the adding in the 1st day that fungal elicitor is cultivated in glossy ganoderma fermentation in the culturing step, then with the glossy ganoderma co-cultivation.
8, a kind of ganoderma lucidum mycelium liquid fermentation method that utilizes fungal elicitor to improve ganoderic acid content according to claim 1, it is characterized in that described glossy ganoderma and fungal elicitor are total to the adding in the 3rd day that fungal elicitor is cultivated in glossy ganoderma fermentation in the culturing step, then with the glossy ganoderma co-cultivation.
9, a kind of ganoderma lucidum mycelium liquid fermentation method that utilizes fungal elicitor to improve ganoderic acid content according to claim 1, it is characterized in that described glossy ganoderma and fungal elicitor are total to the adding in the 5th day that fungal elicitor is cultivated in glossy ganoderma fermentation in the culturing step, then with the glossy ganoderma co-cultivation.
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| CN101928699A (en) * | 2010-04-20 | 2010-12-29 | 山东省农业科学院土壤肥料研究所 | Fermentation process for improving yield of lucid ganoderma laccase |
| CN103392510A (en) * | 2013-08-09 | 2013-11-20 | 南京农业大学 | Method for increasing mycelium ganodenic acid content of ganoderma lucidum liquid in fermentation |
| CN104904491A (en) * | 2014-03-10 | 2015-09-16 | 廉美兰 | Method for improving saponin content in American ginseng adventive roots cultured in reactor with ginseng pathogenic bacterium elicitor |
| CN105002095A (en) * | 2014-04-15 | 2015-10-28 | 大自然生机股份有限公司 | Composite ganoderma strain co-culture method and applications thereof |
| CN106086147A (en) * | 2016-06-30 | 2016-11-09 | 浙江大学 | Fungal elicitor is utilized to induce the method extracting betulic acid from Inonqqus obliquus |
| CN107881120A (en) * | 2017-12-07 | 2018-04-06 | 中国农业科学院麻类研究所 | Ganoderma lucidum mycelium protoplasm with whitening active and preparation method thereof, there are the cosmetics of white-skinned face function |
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| CN108018213A (en) * | 2017-12-01 | 2018-05-11 | 中国农业科学院麻类研究所 | A kind of method for preparing glossy ganoderma mycelium fermentation liquid, fermentate and its application |
| CN110982868A (en) * | 2019-11-29 | 2020-04-10 | 贵州中医药大学 | Co-culture method for improving triterpene content of ganoderma lucidum and application |
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| CN101928699B (en) * | 2010-04-20 | 2012-09-19 | 山东省农业科学院土壤肥料研究所 | Fermentation process for improving yield of lucid ganoderma laccase |
| CN101928699A (en) * | 2010-04-20 | 2010-12-29 | 山东省农业科学院土壤肥料研究所 | Fermentation process for improving yield of lucid ganoderma laccase |
| CN103392510A (en) * | 2013-08-09 | 2013-11-20 | 南京农业大学 | Method for increasing mycelium ganodenic acid content of ganoderma lucidum liquid in fermentation |
| CN104904491A (en) * | 2014-03-10 | 2015-09-16 | 廉美兰 | Method for improving saponin content in American ginseng adventive roots cultured in reactor with ginseng pathogenic bacterium elicitor |
| CN105002095A (en) * | 2014-04-15 | 2015-10-28 | 大自然生机股份有限公司 | Composite ganoderma strain co-culture method and applications thereof |
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| CN108018213A (en) * | 2017-12-01 | 2018-05-11 | 中国农业科学院麻类研究所 | A kind of method for preparing glossy ganoderma mycelium fermentation liquid, fermentate and its application |
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| CN113318014A (en) * | 2021-06-18 | 2021-08-31 | 广州市暨源生物科技有限公司 | Ganoderma lucidum fermentation product, preparation method thereof and application thereof in cosmetics |
| CN113318014B (en) * | 2021-06-18 | 2022-07-05 | 广州市暨源生物科技有限公司 | Ganoderma lucidum fermentation product, preparation method thereof and application thereof in cosmetics |
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