CN102796782A - Use of sodium nitroprusside in ganoderma lucidum submerged fermentation for producing ganoderma lucidum polysaccharide - Google Patents
Use of sodium nitroprusside in ganoderma lucidum submerged fermentation for producing ganoderma lucidum polysaccharide Download PDFInfo
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- CN102796782A CN102796782A CN2012102990632A CN201210299063A CN102796782A CN 102796782 A CN102796782 A CN 102796782A CN 2012102990632 A CN2012102990632 A CN 2012102990632A CN 201210299063 A CN201210299063 A CN 201210299063A CN 102796782 A CN102796782 A CN 102796782A
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- ganoderma lucidum
- sodium nitroprusside
- ganoderan
- submerged fermentation
- glossy ganoderma
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Abstract
The invention discloses a use of sodium nitroprusside in ganoderma lucidum submerged fermentation for producing ganoderma lucidum polysaccharide. After ganoderma lucidum submerged fermentation solution culture lasting for 3-5 days, sodium nitroprusside as an inducer is added into a ganoderma lucidum submerged fermentation solution to induce ganoderma lucidum polysaccharide production in ganoderma lucidum submerged fermentation. Sodium nitroprusside crystals as nitric oxide donors are dissolved in water so that a sodium nitroprusside aqueous solution is formed and is used as an inducer for ganoderma lucidum submerged fermentation used for producing ganoderma lucidum polysaccharide. Through slowly releasing nitric oxide in cells, sodium nitroprusside can promote secondary metabolite ganoderma lucidum polysaccharide synthesis. A result of an experimental research shows that through utilization of an appropriate amount of sodium nitroprusside in a ganoderma lucidum fermentation solution, a ganoderma lucidum polysaccharide yield can be effectively increased by 25 to 160%.
Description
Technical field
The present invention relates to deep glossy ganoderma fermenting technology, particularly Sodium Nitroprusside (SNP) application in deep glossy ganoderma fermenting secondary metabolite ganoderan, to improve the output of ganoderan.
Background technology
Glossy ganoderma [
Ganoderma lucidum(Fr.) Krast (Polyporaceae)] be under the jurisdiction of Aphyllophorales (the aptychus Zoopagales,
Aphyllophorales) Ganodermataceae (
Ganodermataceae) ganoderma (
Ganoderma), have the title of " celestial grass ", " miraculous cure ", be a kind of food medicine dual-purpose fungi, in China, Japan, Korea S and Taiwan long applicating history is arranged.Modern biology has proved and has contained polyose, ucleosides, aminoacid protein class, triterpenes, furans, alkaloids, oils, sterols, organic germanium, mineral ion isoreactivity material in Ganoderma sporophore and the mycelium; Wherein ganoderan is present in the main active ingredient of Ganoderma sporophore, mycelium, spore powder; Have antitumor, anti-ageing, anti-oxidant, AIDS virus resisting, prevent arteriosclerosis and thrombosis, improve hyperlipidemia, improve pharmacological actions widely such as body immunity, analgesia, have very high pharmaceutical use.Therefore, how to utilize the modern biology technology to improve ganoderan output is the important research field of microbiology, Horticulture and fermentation industry always, and pharmaceutical sector and Agricultural Development are had immeasurable value.
At present, ganoderan mainly extracts from Ganoderma sporophore.The artificial culture Ganoderma sporophore production cycle is long, and growth conditions and quality product are wayward.Adopt the submerged fermentation cultured method to obtain ganoderan, have the production cycle shorten widely, system easy to control the quality, main ingredient and wild Ganoderma basically identical, and the polysaccharide content in the fermentation mycelium also is higher than characteristics such as sporophore.It is thus clear that submerged fermentation is a kind of effective way of obtaining ganoderan.Yet the output problem of lower is still a bottleneck of restriction glossy ganoderma deep fermentation large-scale production.A large amount of researchs show, in substratum, add signaling molecule such as certain density fungi exciton, calcium ion, cupric ion, hydrogen peroxide and jasmonic and plant hormone and all can to a certain degree improve ganoderan and ganoderic acid content.Yet the biochemical functions of animal and plant signaling molecule nitrogen protoxide (NO) in edible and medicinal fungi do not launched at present both at home and abroad as yet.
Summary of the invention
The object of the present invention is to provide the application of Sodium Nitroprusside in deep glossy ganoderma fermenting secondary metabolite ganoderan, Sodium Nitroprusside is induced the deep glossy ganoderma fermenting ganoderan as inductor, to improve the output of ganoderan.
According to the fermenting characteristic of Ganderma lucidum strain and the condition test of extensive screening of physiological metabolism characteristic process and system; After finding to adopt the certain density aqueous solution of nitrogen protoxide (NO) donor sodium nitroprusside (SNP) preparation through the filtering bacterium; Certain phase in the glossy ganoderma cell growth adds in the submerged fermentation liquid, can improve the output of ganoderan.
The application of Sodium Nitroprusside in the deep glossy ganoderma fermenting ganoderan is characterized in that: Sodium Nitroprusside is induced the deep glossy ganoderma fermenting ganoderan as inductor, improves the output of ganoderan.
Sodium Nitroprusside induces the specific practice of deep glossy ganoderma fermenting ganoderan to be as inductor; When deep glossy ganoderma fermenting liquid is cultivated 3～5 days; In fermented liquid, adding concentration is the Sodium Nitroprusside aqueous solution of 500mmol/L; The volume ratio of the Sodium Nitroprusside aqueous solution and fermented liquid is 0.1～2%, continues to cultivate after 4～8 days, obtains the tunning that contains ganoderan.
Optimal technical scheme of the present invention is that adding the concentration that accounts for fermentating liquid volume 0.2～1.0% in the fermented liquid is the 500mmol/L Sodium Nitroprusside aqueous solution.
Optimal technical solution of the present invention is that adding the concentration that accounts for fermentating liquid volume 0.2% in the fermented liquid is the 500mmol/L Sodium Nitroprusside aqueous solution.
Principle of the present invention is that the glossy ganoderma cell oxidation that nitrogen protoxide induces submerged fermentation to cultivate is burst out, and promotes the generation of hydrogen peroxide isoreactivity oxygen, and then induces glossy ganoderma to synthesize the secondary metabolite Expression of Related Genes.
The present invention adopt nitrogen protoxide (NO) donor sodium nitroprusside crystal water-soluble be mixed with the Sodium Nitroprusside aqueous solution as the deep glossy ganoderma fermenting secondary metabolite inductor; Sodium Nitroprusside is through slowly releasing NO in cell; Exercise it and promote secondary metabolite synthetic function; Experimental studies results shows, adopts the present invention in Ganoderma fermentation liquid, to add an amount of Sodium Nitroprusside and can effectively improve ganoderan output 25～160%.
Embodiment
The application of Sodium Nitroprusside (SNP) in improving submerged fermentation ganoderan output, concrete grammar is following:
1) compound concentration is the Sodium Nitroprusside (Na of 500mmol/L
2[Fe (CN)
5NO]) aqueous solution;
2) get the glossy ganoderma that refrigerator preserves [
Ganoderma lucidum(Fr.) Krast (Polyporaceae)] the female kind on the fresh yam integrated solid medium slant of mycelium access; Place 28 ℃ of incubators to cultivate 7 days; Mycelia is inserted the comprehensive liquid nutrient medium of yam; In 25～30 ℃ of constant temperature shaking tables, cultivated under 100～160rpm condition 2～5 days, with decollator mycelium pellet is smashed and be the glossy ganoderma seed liquor.
Embodiment one: get the glossy ganoderma seed liquor and fill in the Erlenmeyer flask of 95mL liquid nutrient medium (including glucose 2.0 grams, peptone 1.0 grams, potassium primary phosphate 0.1 gram, sal epsom 0.1 gram) by five of 5% inoculum size accesses; In 28 ℃ of constant temperature shaking tables, when 140rpm cultivates 3 days; Add 0,0.1,0.2,1.0 respectively, 2.0mL concentration is the Sodium Nitroprusside aqueous solution of 500 mmol/L; Continue to cultivate after 6 days, results contain the tunning (fermented liquid) of ganoderan.With fermented liquid with the centrifugal 10min of 5000rpm; Supernatant concentrates in 60 ℃ of rotatory evaporators; Add 4 times of volume absolute ethyl alcohols and leave standstill 48h in 4 ℃ of refrigerators; The centrifugal 10min of 5000rpm must precipitate, and removes Deproteinization with the Sevag method, and dialysing with deionized water at 4 ℃, 24h, 60 ℃ of rotatory evaporators concentrate ,-45 ℃ of vacuum lyophilization 36h.Ganoderan is confirmed in 540 nanometer absorption peaks through the 756MC ultraviolet-visible pectrophotometer, gets 0.065 gram, 0.081 gram, 0.173 gram, 0.132 gram, 0.096 kairine sesame polysaccharide respectively.
Embodiment two: get the glossy ganoderma seed liquor and fill in the Erlenmeyer flask of 95mL liquid nutrient medium (including glucose 2.0 grams, peptone 1.0 grams, potassium primary phosphate 0.1 gram, sal epsom 0.1 gram) by five of 5% inoculum size accesses; In 28 ℃ of constant temperature shaking tables, when 140rpm cultivates 5 days; Add 0,0.1,0.2,1.0 respectively, 2.0mL concentration is the Sodium Nitroprusside aqueous solution of 500 mmol/L; Continue to cultivate after 4 days, results contain the tunning (fermented liquid) of ganoderan.With fermented liquid with the centrifugal 10min of 5000rpm; Supernatant concentrates in 60 ℃ of rotatory evaporators; Add 4 times of volume absolute ethyl alcohols and leave standstill 48h in 4 ℃ of refrigerators; The centrifugal 10min of 5000rpm must precipitate, and removes Deproteinization with the Sevag method, and dialysing with deionized water at 4 ℃, 24h, 60 ℃ of rotatory evaporators concentrate ,-45 ℃ of vacuum lyophilization 36h.Ganoderan is confirmed in 540 nanometer absorption peaks through the 756MC ultraviolet-visible pectrophotometer, gets 0.072 gram, 0.096 gram, 0.186 gram, 0.141 gram, 0.093 kairine sesame polysaccharide respectively.
Embodiment three: get the glossy ganoderma seed liquor and fill in the Erlenmeyer flask of 95mL liquid nutrient medium (including glucose 2.0 grams, peptone 1.0 grams, potassium primary phosphate 0.1 gram, sal epsom 0.1 gram) by five of 5% inoculum size accesses; In 28 ℃ of constant temperature shaking tables, when 140rpm cultivates 4 days; Add 0,0.1,0.2,1.0 respectively, 2.0mL concentration is the Sodium Nitroprusside aqueous solution of 500 mmol/L; Continue to cultivate after 6 days, results contain the tunning (fermented liquid) of ganoderan.With fermented liquid with the centrifugal 10min of 5000rpm; Supernatant concentrates in 60 ℃ of rotatory evaporators; Add 4 times of volume absolute ethyl alcohols and leave standstill 48h in 4 ℃ of refrigerators; The centrifugal 10min of 5000rpm must precipitate, and removes Deproteinization with the Sevag method, and dialysing with deionized water at 4 ℃, 24h, 60 ℃ of rotatory evaporators concentrate ,-45 ℃ of vacuum lyophilization 36h.Ganoderan is confirmed in 540 nanometer absorption peaks through the 756MC ultraviolet-visible pectrophotometer, gets 0.061 gram, 0.076 gram, 0.162 gram, 0.119 gram, 0.087 kairine sesame polysaccharide respectively.
The above only is preferred embodiment of the present invention, is not the present invention is done any pro forma restriction; Any those of ordinary skill in the art; Do not breaking away under the technical scheme scope situation of the present invention; All the method for above-mentioned announcement capable of using and technology contents are made many possible changes and modification to technical scheme of the present invention, or are revised as the equivalent embodiment of equivalent variations.Therefore, every content that does not break away from technical scheme of the present invention, according to technical spirit of the present invention to any simple modification that above embodiment did, be equal to replacement, equivalence changes and modify, all still belong in the scope that technical scheme of the present invention protects.
Claims (4)
1. the application of Sodium Nitroprusside in the deep glossy ganoderma fermenting ganoderan is characterized in that: Sodium Nitroprusside is induced the deep glossy ganoderma fermenting ganoderan as inductor, improves the output of ganoderan.
2. according to the application of the said Sodium Nitroprusside of claim 1 in the deep glossy ganoderma fermenting ganoderan; It is characterized in that: Sodium Nitroprusside induces the specific practice of deep glossy ganoderma fermenting ganoderan to be as inductor; When deep glossy ganoderma fermenting liquid was cultivated 3～5 days, in fermented liquid, adding concentration was the Sodium Nitroprusside aqueous solution of 500mmol/L, and the volume ratio of the Sodium Nitroprusside aqueous solution and fermented liquid is 0.1～2%; Continue to cultivate after 4～8 days, obtain the tunning that contains ganoderan.
3. according to the application of the said Sodium Nitroprusside of claim 2 in the deep glossy ganoderma fermenting ganoderan, it is characterized in that: the volume ratio of the said Sodium Nitroprusside aqueous solution and fermented liquid is 0.2～1.0%.
4. according to the application of the said Sodium Nitroprusside of claim 2 in the deep glossy ganoderma fermenting ganoderan, it is characterized in that: the volume ratio of the said Sodium Nitroprusside aqueous solution and fermented liquid is 0.2%.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109258296A (en) * | 2018-12-05 | 2019-01-25 | 郑涛 | A kind of semicontinuous deep layer fermenting process of high yield Blackfungus polyhexose |
| CN111662939A (en) * | 2020-07-24 | 2020-09-15 | 江苏省中国科学院植物研究所 | Method for promoting burkholderia to synthesize toxoflavin by using signal molecules |
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2012
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| 王慧杰 等: "灵芝多糖液体发酵条件优化", 《食用菌学报》 * |
| 王松华 等: "一氧化氮对灵芝多糖合成的诱导作用", 《2011国际灵芝研究学术会议论文集》 * |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109258296A (en) * | 2018-12-05 | 2019-01-25 | 郑涛 | A kind of semicontinuous deep layer fermenting process of high yield Blackfungus polyhexose |
| CN109258296B (en) * | 2018-12-05 | 2020-11-06 | 江西天元药业有限公司 | Semi-continuous submerged fermentation process for high-yield auricularia auricula polysaccharide |
| CN111662939A (en) * | 2020-07-24 | 2020-09-15 | 江苏省中国科学院植物研究所 | Method for promoting burkholderia to synthesize toxoflavin by using signal molecules |
| CN111662939B (en) * | 2020-07-24 | 2024-02-13 | 江苏省中国科学院植物研究所 | Method for promoting burkholderia to synthesize toxic flavine by using signal molecules |
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Application publication date: 20121128 |