CN105483197A - Method for improving content of triterpene in liquid fermentation of lucid ganoderma - Google Patents
Method for improving content of triterpene in liquid fermentation of lucid ganoderma Download PDFInfo
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- CN105483197A CN105483197A CN201510983745.9A CN201510983745A CN105483197A CN 105483197 A CN105483197 A CN 105483197A CN 201510983745 A CN201510983745 A CN 201510983745A CN 105483197 A CN105483197 A CN 105483197A
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Abstract
The invention relates to a method for improving the content of triterpene in liquid fermentation of lucid ganoderma. The method comprises the following steps: preparation of a CaCl2 water solution and a salicylic acid ethanol solution, liquid culture of hypha, calcium ion and salicylic acid induction, hypha collection as well as extraction and detection of triterpene of the hypha. Calcium ions and salicylic acid are added in a liquid fermentation process of the lucid ganoderma to carry out induction, such that the content of the triterpene in the lucid ganoderma hypha can be remarkably improved, the content of the triterpene in the hypha reaches 48.97mg/g which is improved by 48.26 percent compared with that of a control group, the quality of the lucid ganoderma is effectively improved, and the operation method is simple and low in cost.
Description
Technical field
The present invention relates to a kind of method improving triterpene content in edible fungus liquid fermentation, be specifically related to a kind of method improving triterpene content in Ganoderma lucidum submerged fermentation.
Technical background
Glossy ganoderma [Ganodermalucidum(Curtis:Fr.) P.Karst] Gu is called auspicious grass, have another name called for red sesame, 10000 years gill fungus, red sesame, Ganoderma lucidum seu Japonicumes etc., the classification position of fungi belongs to mycota (Mycota), Basidiomycotina (Basidiomycotina), layer bacterium subclass (Hymenomycetes), Aphyllophorales (Aphyllophorales), polyporaceae (Polyproraceae), Ganoderma (Ganoderma).Ganoderma triterpenoids has good pharmacological action, is mainly manifested in the growth of anticancer, liver protecting, hypotensive, AntiHIV1 RT activity etc.
The quality of glossy ganoderma quality and the content of Ganoderma triterpenoids closely related.The mycelia of the source current production obtaining ganoderma lucidum triterpene compounds mainly mature sporophore and liquid fermenting.The cycle of being extracted the triterpene of glossy ganoderma by liquid fermentation technology is shorter, and simple to operate.Along with the continuous release of ganoderma lucidum prod on market, just require to produce Ganoderma triterpenoids more efficiently.Therefore, explore the content how effectively improving triterpene in Ganoderma lucidum submerged fermentation, be with a wide range of applications.
Summary of the invention
The object of this invention is to provide a kind of method improving triterpene content in Ganoderma lucidum submerged fermentation, meeting the market requirement, also for effective component of glossy ganoderma analysis provides foundation.
A kind of method improving triterpene content in Ganoderma lucidum submerged fermentation of the present invention, operation steps is as follows:
(1) CaCl is prepared
2the aqueous solution and Whitfield's ointment ethanolic soln: use CaCl
2with the sterilized water preparation 10mM calcium ion aqueous solution; With Whitfield's ointment and ethanol, prepare 150 μMs of Whitfield's ointment ethanolic solns;
(2) mycelia liquid culture: get in 8 ferfas silk access 100mL liquid PDA from the flat board covering with Ganoderma lucidum mycelium with 9mm punch tool, 28 DEG C, 6-7 days cultivated by 150rpm shaking table, and then quiescent culture 5-6 days; Described liquid PDA formula is: 200g potato boils leach liquor, 20g glucose, the constant volume that adds water to 1000mL, 121 DEG C of sterilizing 30min;
(3) calcium ion and Induced by Salicylic Acid: the calcium ion aqueous solution 1-5mL and the Whitfield's ointment ethanolic soln 0.5-1.5mL that get step (1), mixing, filter membrane degerming method is adopted to join in the PDA nutrient solution of step (2) inoculation culture, induction process 1-2 days in 28 DEG C of biochemical cultivation cases;
(4) mycelia is collected: collect the mycelia after step (3) induction process, and removing inoculation block, with distilled water wash 3-5 time, be placed in 60 DEG C of baking ovens and dry to constant weight;
(5) mycelia triterpene extracts and detects: after getting the mycelia alcohol at normal temperature lixiviate 2h that step (4) dries, the ultrasonic 0.5h of 150W, and 60 DEG C rotate evaporates to dryness, use dissolve with methanol constant volume, and adopts Vanillin-ice acetic acid method to detect triterpene content.
Advantage of the present invention and usefulness: in Ganoderma lucidum submerged fermentation process, add calcium ion and Whitfield's ointment is induced, can significantly improve the content of triterpene in Ganoderma lucidum mycelium, improve the quality of glossy ganoderma, and cost simple to operate is low.Ganoderma triterpenoids is pharmacological component main in glossy ganoderma, and by adding calcium ion and salicylic induction, effectively can improve the content of triterpene in Ganoderma lucidum submerged fermentation, the triterpene content of mycelia reaches 48.97mg/g, higher than control group by 48.26%.
Accompanying drawing explanation
Fig. 1 is the triterpene content schematic diagram of different incubation time mycelia.In figure, X-coordinate represents the number of days of cultivation, and ordinate zou represents triterpene content mg/g.
Fig. 2 adds calcium ion and Whitfield's ointment to affect schematic diagram to Ganoderma triterpenoids content.In figure, CK is contrast, and Ca+SA represents that carrying out induction with method interpolation calcium ion of the present invention and Whitfield's ointment processes; * represent and contrast that there were significant differences, P < 0.05; Ordinate zou represents triterpene content mg/g.Do not add calcium ion and salicylic mycelia, triterpene content is 33.03mg/g; Interpolation calcium ion and salicylic mycelia triterpene content are 48.97mg/g, higher than control group by 48.26%.
Embodiment
In order to fully disclose the method for the content of triterpene in raising Ganoderma lucidum submerged fermentation of the present invention, be illustrated below in conjunction with example.
Embodiment 1: a kind of method improving triterpene content in Ganoderma lucidum submerged fermentation.Comprise the following steps:
(1) CaCl is prepared
2the aqueous solution and Whitfield's ointment ethanolic soln: use CaCl
2with the sterilized water preparation 10mM calcium ion aqueous solution; With Whitfield's ointment and ethanol, prepare 150 μMs of Whitfield's ointment ethanolic solns;
(2) mycelia liquid culture: the ganoderma strain capable adopting market to buy, accesses the flat board of 90mm, after flat board covers with mycelia after PDA inclined-plane activates, get in 8 ferfas silk access 100mL liquid PDA with 9mm punch tool, 28 DEG C, 150rpm shaking table cultivates 7 days, and then quiescent culture 5 days; Described liquid PDA formula is: 200g potato boils leach liquor, 20g glucose, the constant volume that adds water to 1000mL, 121 DEG C of sterilizing 30min; The triterpene content of different incubation time mycelia as shown in Figure 1; When cultivation was by the 13rd day, in mycelia, triterpene content reaches maximum value 33.03mg/g.
(3) calcium ion and Induced by Salicylic Acid: the calcium ion aqueous solution 1mL and the Whitfield's ointment ethanolic soln 1mL that get step (1), mixing, adopt the degerming method of filter membrane, namely 1mL asepsis injector via hole diameter is adopted to be 0.22 μm of sterilized organic filter, join in the PDA nutrient solution of step (2) inoculation culture, in 28 DEG C of biochemical cultivation cases, induce process 1 day.On the impact of Ganoderma triterpenoids content as shown in Figure 2, interpolation calcium ion and salicylic mycelia triterpene content are 48.97mg/g, higher than control group by 48.26% for interpolation calcium ion and Whitfield's ointment.
(4) mycelia is collected: collect the mycelia after step (3) induction process, and removing inoculation block, with distilled water wash 5 times, be placed in 60 DEG C of baking ovens and dry to constant weight;
(5) mycelia triterpene extracts and detects: the mycelia 0.2g taking oven dry, with 30mL85%(v/v) after alcohol at normal temperature lixiviate 2h, the ultrasonic 0.5h of 150W, 60 DEG C rotate evaporates to dryness, are finally settled to 25mL with dissolve with methanol.Vanillin-ice acetic acid method is adopted to detect triterpene content.
Described Vanillin-ice acetic acid method detects triterpene content: get the sample solution of 0.2mL in tool plug test tube, evaporate to dryness in 60 DEG C of water-baths, add the 5% Vanillin-Glacial acetic acid (now with the current) of 0.4mL and the perchloric acid of 1mL, shake up the Glacial acetic acid adding 5mL after rear 60 DEG C of water-bath 15min. are cooled to room temperature again, shake up rear room temperature and leave standstill 0.5h, 546nm carries out OD value and detects.
Embodiment 2: a kind of method improving triterpene content in Ganoderma lucidum submerged fermentation.Comprise the following steps:
(1) CaCl is prepared
2the aqueous solution and Whitfield's ointment ethanolic soln: use CaCl
2with the sterilized water preparation 10mM calcium ion aqueous solution; With Whitfield's ointment and ethanol, prepare 150 μMs of Whitfield's ointment ethanolic solns;
(2) mycelia liquid culture: the ganoderma strain capable adopting market to buy, accesses the flat board of 90mm, after flat board covers with mycelia after PDA inclined-plane activates, get in 8 ferfas silk access 100mL liquid PDA with 9mm punch tool, 28 DEG C, 150rpm shaking table cultivates 6 days, and then quiescent culture 6 days;
(3) calcium ion and Induced by Salicylic Acid: the calcium ion aqueous solution 5mL and the Whitfield's ointment ethanolic soln 1.5mL that get step (1), mixing, adopt filter membrane degerming method join step (2) inoculation culture PDA nutrient solution in, induce process 2 days in 28 DEG C of biochemical cultivation cases.
(4) mycelia is collected: collect the mycelia after step (3) induction process with gauze, and removing inoculation block, with distilled water wash 3 times, be placed in 60 DEG C of baking ovens and dry to constant weight.
(5) mycelia triterpene extracts and detects: the mycelia 0.2g taking oven dry, with 30mL85%(v/v) after alcohol at normal temperature lixiviate 2h, the ultrasonic 0.5h of 150W, 60 DEG C rotate evaporates to dryness, finally use dissolve with methanol constant volume 25mL, and adopt Vanillin-ice acetic acid method to detect triterpene content.
The foregoing is only preferred embodiment of the present invention, all equalizations done according to the present patent application the scope of the claims change and modify, and all should belong to covering scope of the present invention.
Claims (3)
1. improve a method for triterpene content in Ganoderma lucidum submerged fermentation, it is characterized in that operation steps is as follows:
(1) CaCl is prepared
2the aqueous solution and Whitfield's ointment ethanolic soln: use CaCl
2with the sterilized water preparation 10mM calcium ion aqueous solution; With Whitfield's ointment and ethanol, prepare 150 μMs of Whitfield's ointment ethanolic solns;
(2) mycelia liquid culture: get in 8 ferfas silk access 100mL liquid PDA from the flat board covering with Ganoderma lucidum mycelium with 9mm punch tool, 28 DEG C, 6-7 days cultivated by 150rpm shaking table, and then quiescent culture 5-6 days; Described liquid PDA formula is: 200g potato boils leach liquor, 20g glucose, the constant volume that adds water to 1000mL, 121 DEG C of sterilizing 30min;
(3) calcium ion and Induced by Salicylic Acid: the calcium ion aqueous solution 1-5mL and the Whitfield's ointment ethanolic soln 0.5-1.5mL that get step (1), mixing, filter membrane degerming method is adopted to join in the PDA nutrient solution of step (2) inoculation culture, induction process 1-2 days in 28 DEG C of biochemical cultivation cases;
(4) mycelia is collected: collect the mycelia after step (3) induction process, and removing inoculation block, with distilled water wash 3-5 time, be placed in 60 DEG C of baking ovens and dry to constant weight;
(5) mycelia triterpene extracts and detects: after getting the mycelia alcohol at normal temperature lixiviate 2h that step (4) dries, the ultrasonic 0.5h of 150W, and 60 DEG C rotate evaporates to dryness, use dissolve with methanol constant volume, and adopts Vanillin-ice acetic acid method to detect triterpene content.
2. a kind of method improving triterpene content in Ganoderma lucidum submerged fermentation according to claim 1, it is characterized in that described mycelia liquid culture: get in 8 ferfas silk access 100mL liquid PDA with 9mm punch tool from the flat board covering with Ganoderma lucidum mycelium, 28 DEG C, 150rpm shaking table cultivates 7 days, and then quiescent culture 5 days.
3. a kind of method improving triterpene content in Ganoderma lucidum submerged fermentation according to claim 1, it is characterized in that described calcium ion and Induced by Salicylic Acid: get calcium ion aqueous solution 1mL and Whitfield's ointment ethanolic soln 1mL, mixing, adopt the degerming method of filter membrane to join in the PDA nutrient solution of inoculation culture, in 28 DEG C of biochemical cultivation cases, induce process 1 day.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105506049A (en) * | 2016-02-02 | 2016-04-20 | 华中农业大学 | Method for increasing content of intracellular ganoderma lucidum triterpenes during liquid fermentation of ganoderma lucidum mycelia |
| CN106538237A (en) * | 2016-10-18 | 2017-03-29 | 福建农林大学 | A kind of raising Ganoderma sporophore yield and polysaccharide, the processing method of triterpene content |
| CN109168964A (en) * | 2018-11-28 | 2019-01-11 | 山东农业大学 | A kind of method of ganodenic acid content in raising ganoderma lucidum fruitbody |
| CN109938040A (en) * | 2019-04-22 | 2019-06-28 | 山东省农业科学院生物技术研究中心 | A method of the anti-root rot of peanut is improved using salicylic acid and calcium |
| 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 |
-
2015
- 2015-12-24 CN CN201510983745.9A patent/CN105483197B/en active Active
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105506049A (en) * | 2016-02-02 | 2016-04-20 | 华中农业大学 | Method for increasing content of intracellular ganoderma lucidum triterpenes during liquid fermentation of ganoderma lucidum mycelia |
| CN105506049B (en) * | 2016-02-02 | 2018-11-27 | 华中农业大学 | A method of improving the ganodenic acid content intracellular of ganoderma lucidum mycelium liquid fermentation |
| CN106538237A (en) * | 2016-10-18 | 2017-03-29 | 福建农林大学 | A kind of raising Ganoderma sporophore yield and polysaccharide, the processing method of triterpene content |
| CN109168964A (en) * | 2018-11-28 | 2019-01-11 | 山东农业大学 | A kind of method of ganodenic acid content in raising ganoderma lucidum fruitbody |
| CN109938040A (en) * | 2019-04-22 | 2019-06-28 | 山东省农业科学院生物技术研究中心 | A method of the anti-root rot of peanut is improved using salicylic acid and calcium |
| CN109938040B (en) * | 2019-04-22 | 2021-10-01 | 山东省农业科学院生物技术研究中心 | Method for improving peanut root rot resistance by using salicylic acid and calcium |
| 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 |
| CN110982868B (en) * | 2019-11-29 | 2023-09-19 | 贵州中医药大学 | Co-culture method for improving triterpene content of ganoderma lucidum and application thereof |
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