CN101851356A - Shitake mushroom extract and application thereof - Google Patents
Shitake mushroom extract and application thereof Download PDFInfo
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- CN101851356A CN101851356A CN201010198551A CN201010198551A CN101851356A CN 101851356 A CN101851356 A CN 101851356A CN 201010198551 A CN201010198551 A CN 201010198551A CN 201010198551 A CN201010198551 A CN 201010198551A CN 101851356 A CN101851356 A CN 101851356A
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Abstract
The invention relates to a shitake mushroom extract. The shitake mushroom is used as raw materials, and the extraction process comprises a step of ultrasonic extraction. The shitake mushroom extract is not extracted by the traditional enzymolysis method for extracting lentinan. Due to the adoption of the ultrasonic extraction, the time is shortened to 1/3 to 1/4 of that of the traditional method, the temperature is reduced by 30 to 40 DEG C, and the yield is increased by 5 to 30 percent. The method has the advantages of simple technical process, less procedure, short extraction time, resource conservation and low production cost; and the prepared lentinan has high content of parent medicament and high economic value, is favorable for mass production and has great development space.
Description
Technical field
The invention belongs to the female medicine of the plant-sourced field of Organic farming, specifically is female medicine of a kind of agricultural lentinan and preparation method thereof.
Background technology
Because the use of high malicious high residue chemical pesticide makes the pollution of water source, soil, livestock etc. serious day by day, serious threat existent environment of people.Current protection environment and food safety more and more cause people's attention.The effective active matter that utilization is modern biological, the chemical industry technology is sought new highly-effective low-residue, the inexorable trend that to replace high malicious high residue compound be human social development.
The viroses of plant are of a great variety, and harm is serious, the tens billion of approximately units of financial loss that grain-production, garden crop, cash crop are caused.China prevents and treats virus disease and adopts chemical process more at present, and domestic this type of disease obvious results product of control that lacks demands researching and developing a kind of novel more efficiently product urgently.
Lentinan is isolating a kind of polysaccharide from the edible mushrooms mushroom.Through it is carried out the total composition analysis, the main moiety of lentinan is mainly the mushroom polysaccharide, and monomer whose is made up of 7 glucose.Lentinan is a kind of material that can enhancing body suppresses various bacteriums, virus and parasitic ability.
Lentinan is mainly used in field of medicaments at present, its be used for medical aspect have anti-lipid, antitumor, regulate that immunity, antiplatelet are condensed, effect such as antiviral, and good effect is arranged in clinical.
In 20 end of the centurys, lentinan begins to be applied to agriculture aspect, and it has good restraining and killing action for virus, can strengthen the resistance against diseases of plant, reaches the effect of volume increase, raising plant quality simultaneously.
Lentinan can reduce virus on the one hand to the harm that plant brought, and can improve the resistance against diseases of plant on the other hand again, therefore has comparatively extensive applicability.Can be used for preventing and treating the virus disease of various crop such as paddy rice, tomato, capsicum, cucumber, tobacco, potato, flowers.
In the pertinent literature of present existing extraction lentinan, patent and the report, relevant for using the mushroom root to extract the method for preparing lentinan.The lentinan that this method is extracted is a pressed powder, is used for medicine and food aspect.In addition, also have patent to be to use Lentinus Edodes fungus chaff and handle base of a fruit extract to prepare biological pesticide and technology thereof, this patent is to use the Lentinus Edodes fungus chaff and the handle base of a fruit thereof to carry out enzymolysis, deactivation and concentrate making compound formulation, and what this patent was mainly set forth is the technology of preparation biological pesticide.In the patent No. is that the main raw material that the preparation technology of the female medicine of preparation lentinan in one described in 2009101192575.8 adopts is the mushroom rod, by the mushroom rod is pulverized, immersion, enzymolysis, deactivation, alcohol analyse, filter, dilute the mushroom polysaccharide mother liquor that is prepared from.And adopt the mushroom fruiting body and the handle base of a fruit thereof to use ultrasonic extraction equipment to extract, and its as fertilizer sources can be used, this type of invents the domestic relevant patent that yet there are no.
Summary of the invention
Purpose of the present invention provide a kind of in Organic farming is produced sterilization effect good, low toxicity, low residue and the Shitake Mushroom P.E that can as fertilizer sources uses.
The invention provides a kind of Shitake Mushroom P.E, it is a raw material with the mushroom, comprises the step of supersound extraction in leaching process.
The time of described supersound extraction is 20～40 minutes.
Preferably, the used extractor of supersound extraction is a Continuous Countercurrent Extraction equipment.
Wherein, before supersound extraction, the mushroom raw material is pulverized and soaked, after supersound extraction, carry out alcohol and analyse.
Described pulverizing is for to be crushed to granularity between 60～120 orders with mushroom.
After the pulverizing, with the hot-water soak of 5～20 times of volumes 1～5 hour, preferred 4 hours; Soaking temperature remains on 60～70 ℃.
It is that volume percent is 60～80% ethanol that alcohol is analysed used alcohol, and consumption is 2-4 times of volume of supersound extraction liquid.
The alcohol time of analysing is 3～6 hours, and pure eutectoid temperature is controlled at 0 ℃～5 ℃, and the lye after alcohol is analysed carries out concentrating under reduced pressure.
Particularly, Shitake Mushroom P.E of the present invention prepares by following step:
1) the mushroom raw material pulverizing is crushed to granularity between 60～120 orders, uses the hot-water soak 1～5 hour of 5～20 times of volumes then, preferred 4 hours; Soaking temperature remains on 60～100 ℃;
2) soak solution that step 1 is obtained carries out supersound extraction, and the time is 20～40 minutes;
3) adding volume percent is that 60～80% ethanol carries out alcohol and analyses, and consumption is 2-4 times of volume of supersound extraction liquid, and the alcohol time of analysing is 3～6 hours, and pure eutectoid temperature is controlled at 0 ℃～5 ℃, and the lye after alcohol is analysed carries out concentrating under reduced pressure.
The present invention further provides above-mentioned Shitake Mushroom P.E is used for preventing and treating the agricultural chemicals of the viroses of plant in preparation application.
Shitake Mushroom P.E of the present invention is called the female medicine of lentinan, and its main active ingredient is a lentinan, and content can reach 1%～90%, substantially more than 10%.
Shitake Mushroom P.E of the present invention and preparation method thereof has following beneficial effect:
(1) the present invention uses mushroom fruiting body and the handle base of a fruit thereof the main raw material as mushroom polysaccharide mother liquor, and the raw material natural environmental-protective that is adopted very suitablely is processed into single or compound preparation and fertilizer with it, is applicable to the disease control in the Organic farming production process;
(2) the present invention adopts the raw material of mushroom as the female medicine of agricultural lentinan, and cost is low, and preparation technology is easy, good product quality, and the female pharmaceutically active ingredient content of lentinan height, content can reach 1%～90%, and traditional technology then is difficult to reach more than 1%;
(3) the female medicine of the agricultural lentinan of the present invention's employing is paid attention to the control to the starting material soaking temperature in preparation process, and adopts the extraction of continuous countercurrent supersound extraction equipment to prepare the female medicine of agricultural lentinan; Ultrasonic wave extraction is a kind of novel extractive technique, is applied to the extraction of vegetable drug composition, can reduce extraction time greatly, improve extraction rate, reduce the use of solvent, traditional static is extracted needs several hours to tens hours, and ultrasonic extraction at most only needs dozens of minutes; In preparation process, prolong alcohol in addition and analysed the time, greatly improved the content of the female the effective elements of the medicine of lentinan, saved cost, improved production efficiency.
In sum, the present invention has abandoned the enzyme solution of traditional extraction lentinan, adopts supersound extraction, and its technical process is simple, and operation is few, and extraction time is short, economizes on resources, and production cost is lower, the prepared female medicine content of the lentinan height that goes out.And the raw material natural environmental-protective of this process using, nontoxic, noresidue, the female medicine of the prepared agricultural lentinan that goes out is a kind of environmentally friendly product to the person poultry harmless.In addition, by telling test, contain two kinds of effective constituents of lentinan and amino acid by the female medicine of the agricultural lentinan of this prepared simultaneously.Therefore, this mother's medicine not only has the effect that antivirus action increases crop alimentary, volume increase in addition, can be used as preparation and prevents and treats the plant induced resistance agent and the fertilizer of the viroses of plant, has very big agricultural value and economic benefit.
Embodiment
Following examples are used to illustrate the present invention, but are not used for limiting the scope of the invention.
Embodiment 1
Take by weighing the 60kg mushroom, pulverize the back and throw to bubble material still, add 60 ℃～70 ℃ warm water 300kg, soaked 1 hour.Stir constantly, obtain soak solution; Utilize supersound extraction equipment supersound extraction 40 minutes then; In the extracting solution suction reactor after filtering, adds 4 times of volumes 60% (volume percent) industrial alcohol, lead to saturated cooling salt solution in the chuck and begin alcohol and analyse, stir after 30 minutes, temperature is strict controlled in 0 ℃～5 ℃.After alcohol was analysed 4 hours, concentrating under reduced pressure was made the female soup body of lentinan of 10% (mass percent concentration).
Embodiment 2
Take by weighing the 20kg mushroom, after the pulverizing it is thrown to bubble material still, add 60 ℃～70 ℃ warm water 400kg, soaked 3 hours, stir constantly, obtain soak solution; Utilize supersound extraction equipment supersound extraction 20 minutes then, after the filtration, be evacuated in the reactor, add 2 times of volume 80% industrial alcohols, logical saturated cooling salt solution begins alcohol and analyses in the chuck, stirs after 30 minutes, and temperature is strict controlled in 0 ℃～5 ℃.After alcohol was analysed 6 hours, concentrating under reduced pressure was made the female soup body of lentinan that active constituent content is 15% (mass percent concentration).
Embodiment 3
Take by weighing the 30kg mushroom, after the pulverizing it is thrown to bubble material still, add 60 ℃～70 ℃ warm water 300kg, soaked 4 hours, stir constantly, obtain soak solution; Utilized the ultrasonic device supersound extraction then 30 minutes, and after the filtration, be evacuated in the reactor, add 3 times of volume 60% industrial alcohol 200kg, logical saturated cooling salt solution begins alcohol and analyses in the chuck, stirs after 30 minutes, and temperature is strict controlled in 0 ℃～5 ℃.After alcohol was analysed 4 hours, concentrating under reduced pressure was made the female soup body of lentinan that active constituent content is 20% (mass percent concentration).
Embodiment 4
Take by weighing the 60kg mushroom, after the pulverizing it is thrown to bubble material still, add 60 ℃～70 ℃ warm water 300kg, soaked 5 hours, stir constantly, obtain soak solution; Utilized the ultrasonic device supersound extraction then 40 minutes, and after the filtration, be evacuated in the reactor, add 4 times of volume 80% industrial alcohol 200kg, logical saturated cooling salt solution begins alcohol and analyses in the chuck, stirs after 30 minutes, and temperature is strict controlled in 0 ℃～5 ℃.After alcohol was analysed 5 hours, concentrating under reduced pressure was made the female soup body of lentinan that active constituent content is 25% (mass percent concentration).
Experimental example 1
The female medicine of embodiment 1 gained lentinan is added water to be diluted and is prepared into the lentinan aqua that lentinan content is 1% (diluting 10 times by volume).
Because it is tomato virus disease is a tomato main disease at present, especially serious with autumn tomato, delayed culture in autumn tomato.At present, the tomato virus disease morbidity is general, and it is serious to cause harm, and difficulty of prevention and cure is big, does not almost have output even total crop failure after the plant morbidity.Tomato virus disease has become one of important disease that threatens tomato production.It mainly infects virus and mainly contains tobacco mosaic virus (TMV) (TMV) and cucumber mosaic virus (CMV).Tobacco mosaic virus (TMV) host is extensive, can infect 200 various plants, and the tomato virus disease morbidity that causes is commonplace, and general medicament is to the tomato virus disease poor effect.
Test method has two kinds:
(1) whole strain method is measured embodiment 1 gained 1% lentinan activity
Method is as follows: plant common three lives cigarette, plant is long during to 8 true leaves, keeps 4-7 sheet true leaf, inoculate 13.1 μ g/mL tobacco mosaic virus disease poison after, sprayed one time medicament every 7 days, every processing repeats for 3 times.Control treatment is set.Not inoculation and newborn blade are got in dispenser after 14 days, measure medicament to the TMV inhibiting rate, and experiment is finished in the insect protected greenhouse.
Medicament is measured TMV inhibition of proliferation rate
Be cushioned the good sample of liquid dilution process with tobacco mosaic virus disease poison bag, join in the elisa plate well every hole 200 μ l.Blank, negative control and positive control are set simultaneously.Hatch 3h for 37 ℃ in incubator.After adding the anti-clear TMV-lgG of 10 μ l/mL rabbits, hatch 3h in 37 ℃.Second antibody is the mouse-anti rabbit of horseradish peroxidase-labeled, and substrate is an o-phenylenediamine solution.Read the light absorption value of 490nm wavelength with microplate reader, use the OD value representation.Handle light absorption value with each and calculate medicament the TMV inhibiting rate.
(2) withered spot method is measured the 1% lentinan aqua activity that the dilution of embodiment 1 gained lentinan obtains.
Get the healthy young tomato seedling of cultivating in the flowerpot, adopt half leaf method to inoculate, blade is cut into two along master pulse.Contrast is set, six processing, three repetitions.After inoculating 13.1 μ l/mL tobacco mosaic virus disease poison, immediately medicament is sprayed on the surface of half leaf, be placed on 24 ℃ of incubators and cultivate, carry out the withered spot number statistics when scab is counted easily.Control treatment uses fresh water spraying at half leaf surface.
Inhibiting rate=1-handles withered spot number/contrast withered spot number
The result is as follows:
The whole strain method of table 1 is measured 1% lentinan activity
The whole strain measurement result of tobacco shows that embodiment 1 gained lentinan aqua has significant inhibitory effect to TMV, and its EC50 is 121.3mg/L.
The withered spot method of table 2 is measured 1% lentinan activity
Withered spot method measurement result shows that 1% lentinan aqua can effectively suppress TMV, and its EC50 is 127.8mg/L.
Test has been measured the inhibition effect of 1% lentinan to TMV by whole strain method of tobacco and withered spot method, and the result shows that 1% lentinan can effectively suppress TMV.
Experimental example 2
The female medicine of embodiment 2-4 gained lentinan is added water to be diluted and is prepared into the lentinan aqua that lentinan content is 1% (dilution by volume).
According to the identical mode of embodiment 1, it is as follows to utilize whole strain method to measure 1% lentinan activity test method measurement result:
Embodiment 2-4 gained 1% lentinan aqua has significant inhibitory effect to TMV, and its EC50 is respectively 120.9mg/L, 122.3mg/L and 123.6mg/L.
Utilize withered spot method to measure the active result of 1% lentinan and show that embodiment 2-4 gained 1% lentinan aqua can effectively suppress TMV, its EC50 is respectively 126.9mg/L, 128.6mg/L and 129.4mg/L.
Test has been measured the inhibition effect of 1% lentinan to TMV by whole strain method of tobacco and withered spot method, and the result shows that 1% lentinan can effectively suppress TMV.
Claims (10)
1. a Shitake Mushroom P.E is characterized in that, it is a raw material with the mushroom, comprises the step of supersound extraction in leaching process.
2. Shitake Mushroom P.E according to claim 1 is characterized in that, the time of described supersound extraction is 20～40 minutes.
3. Shitake Mushroom P.E according to claim 1 and 2 is characterized in that, the used extractor of supersound extraction is a Continuous Countercurrent Extraction equipment.
4. Shitake Mushroom P.E according to claim 1 is characterized in that, before supersound extraction the mushroom raw material is pulverized and is soaked, and carries out alcohol and analyse after supersound extraction.
5. Shitake Mushroom P.E according to claim 4 is characterized in that described pulverizing is for to be crushed to granularity between 60～120 orders with mushroom.
6. Shitake Mushroom P.E according to claim 4 is characterized in that, after the pulverizing, with the hot-water soak of 5～20 times of volumes 1～5 hour, soaking temperature remained on 60～70 ℃.
7. Shitake Mushroom P.E according to claim 4 is characterized in that, it is that volume percent is 60～80% ethanol that alcohol is analysed used alcohol, and consumption is 2-4 times of volume of supersound extraction liquid.
8. Shitake Mushroom P.E according to claim 7 is characterized in that, the alcohol time of analysing is 3～6 hours, and pure eutectoid temperature is controlled at 0 ℃～5 ℃, and the lye after alcohol is analysed carries out concentrating under reduced pressure.
9. prepare the method for each described Shitake Mushroom P.E of claim 1～8, it is characterized in that, comprise the step of supersound extraction.
10. comprise each described Shitake Mushroom P.E of claim 1～8 is used for preventing and treating the agricultural chemicals of the viroses of plant in preparation application.
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