CN103329733A - Methods for preparing entomogenous fungus conjugate and extracting polysaccharides of entomogenous fungus conjugate - Google Patents
Methods for preparing entomogenous fungus conjugate and extracting polysaccharides of entomogenous fungus conjugate Download PDFInfo
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- CN103329733A CN103329733A CN2013102483859A CN201310248385A CN103329733A CN 103329733 A CN103329733 A CN 103329733A CN 2013102483859 A CN2013102483859 A CN 2013102483859A CN 201310248385 A CN201310248385 A CN 201310248385A CN 103329733 A CN103329733 A CN 103329733A
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Abstract
The invention discloses a method for preparing an entomogenous fungus conjugate from probiotics parasitic centipedes and a method for extracting the polysaccharides of the entomogenous fungus conjugate. The method for preparing the entomogenous fungus conjugate comprises the following steps of (1) culturing cordyceps sinensis; and (2) preparing the conjugate, namely mixing fungus liquid with the centipedes and culturing the mixture to obtain the conjugate of the centipedes and the cordyceps sinensis, and determining the polysaccharides. The method for extracting the polysaccharides of the entomogenous fungus conjugate comprises the steps of (1) dissolution; (2) extraction; and (3) washing, re-concentration, freeze-drying and crushing, so that the polysaccharides of the entomogenous fungus conjugate can be obtained. According to the technical scheme, a great quantity of polysaccharide substances can be generated by the obtained entomogenous fungus conjugate, electrically stimulated centipede bodies can be developed and utilized, the polysaccharide content can be improved, and the economic value of the centipede bodies is favorably improved.
Description
Technical field
The present invention relates to the application of the livings fungi of benefit, be specifically related to a kind ofly prepare the entomogenous fungi bond and extract the method for this bond polysaccharide with probio parasitism centipede.
Background technology
Centipede all has distribution on both sides of the Changjiang River in China, and traditional medicine is used for existing two over thousands of years history of prevention and treatment of diseases with centipede, and the medicinal beginning sees Shennong's Herbal.The centipede head has a pair of jaw foot to contain poison gland can secrete venom, and these venom thrust by posion jaw when predation on the one hand and venom is injected in the prey body, make it paralysis even dead, and these venom help to strengthen the animal defense ability on the other hand.Yet these venom are applied to find after the medical research: these venom have maincenter inhibition and anticonvulsion, improve microcirculation, prevent myocardial damage, reduce blood viscosity, promote the secretion of gastrointestinal motility and digestive ferment, the anti-ageing effect of waiting for a long time.Centipede is got the topic that poison has become everybody to be concerned about thereupon, and in early days centipede being got poison all is the huge legendary turtle foot that adopts the clip centipede, grind then, and water or buffer solution extraction, only can the say so extract of centipede jaw foot of these extracts, its purity is not high.Subsequently, people adopt electrical stimulation method to get poison, and this method can repeatedly be carried out the toxin extraction, and purity is higher relatively.But the animal immune ability descends after repeatedly stimulating, and causes the mortality phenomenon, causes a large amount of spoils to need to handle.These polypides use its standard content to lack as the Chinese medicine toxin, and the deep level development of spoil and utilization become to press for deals with problems.Benefit is given birth to the fungi autoeciousness polypide, transforms the polypide material and forms, and the benefit materials of generation more report, and is applied to clinical practice, prepares stiff silkworm as silkworm larva, the Cordyceps sinensis that the bat moth larvae forms, and the cicada larva forms cicada fungus.These have all realized combination between the entomogenous fungi, produce polysaccharose substance after the living fungi of these benefits of this process is decomposed polypide, and this material is significant for strengthening immune function of human body.Thus, the present invention gets the polypide processing of poison back for centipede an effective treating method is provided, and this method can increase the polypide polyoses content.
Summary of the invention
The object of the invention is to overcome the shortcoming and defect that prior art exists, and provides a kind of and prepares the entomogenous fungi bond and extract the method for this bond polysaccharide with probio parasitism centipede.This can produce a large amount of polysaccharose substances in conjunction with product, and the development and use of centipede polypide improve polyoses content after the realization electro photoluminescence, help to improve the economic worth of polypide.
The object of the present invention is achieved like this:
The method for preparing the entomogenous fungi bond with the parasitic centipede of probio of the present invention is:
(1), the cultivation of aweto: with aweto toothpick picking, put into the activation of PDY liquid nutrient medium, the preparation spore is collected spore, configuration bacterium liquid;
(2), the preparation of bond: with bacterium liquid and centipede polypide number according to 5～60ml: 200～300 ratio, with watering can bacterium liquid evenly being sprayed at polypide surface or polypide was immersed in the bacterium liquid after 1～3 minute, be placed in 15～25 ℃ the environment, cultivated 7～15 days, the beginning blackening appears in polypide after 3 days, and mycelia begins the white hypha body after 5 days, be covered with polypide after 7 days, took out polypide in 10 days, natural seasoning namely gets centipede and aweto in conjunction with product; Carry out polysaccharide determination.
The extracting method of entomogenous fungi bond polysaccharide of the present invention may further comprise the steps:
(1), dissolving: get the bacterium powder of the entomogenous fungi bond of 50g drying, by the bacterium powder: water=1: 25～35 (mass ratio) adds deionized water 3L;
(2), extract: after 2h are extracted in 95 ℃ of water-baths, 5000r/min, centrifugal 8～12min gets supernatant, rotary evaporation is concentrated into 1/4 of original volume;
(3), washing: the water extract after concentrating, the absolute ethyl alcohol that adds 4 times while stirring, leave standstill 8～12h, the centrifugal 8～12min of 5000r/min, abandoning supernatant after fully stirring, precipitation is washed 2 times with absolute ethyl alcohol, acetone, ether successively, deionized water redissolution precipitation with minimum volume concentrates freeze-drying again, pulverize, namely get the polysaccharide of polypide bond.
Because the present invention has taked technique scheme, the entomogenous fungi bond that obtains can produce a large amount of polysaccharose substances, and the development and use of centipede polypide improve polyoses content after the realization electro photoluminescence, help to improve the economic worth of polypide.
Description of drawings
Fig. 1 is entomogenous fungi bond of the present invention.
Embodiment
The method for preparing the entomogenous fungi bond with the parasitic centipede of probio of the present invention is:
(1), the cultivation of aweto: with aweto toothpick picking, put into the activation of PDY liquid nutrient medium, the preparation spore is collected spore, configuration bacterium liquid;
(2), the preparation of bond: with bacterium liquid and centipede polypide number according to 5～60ml: 200～300 ratio, with watering can bacterium liquid evenly being sprayed at polypide surface or polypide was immersed in the bacterium liquid after 1～3 minute, be placed in 15～25 ℃ the environment, cultivated 7～15 days, the beginning blackening appears in polypide after 3 days, and mycelia begins the white hypha body after 5 days, be covered with polypide after 7 days, took out polypide in 10 days, natural seasoning namely gets centipede and aweto in conjunction with product; Carry out polysaccharide determination.
Above-mentioned aweto can be bought by the microorganism resource storehouse and obtain.
The extracting method of entomogenous fungi bond polysaccharide of the present invention may further comprise the steps:
(1), dissolving: get the bacterium powder of the entomogenous fungi bond of 50g drying, by the bacterium powder: water=1: 25～35 (mass ratio) adds deionized water 3L;
(2), extract: after 2h are extracted in 95 ℃ of water-baths, 5000r/min, centrifugal 8～12min gets supernatant, rotary evaporation is concentrated into the l/4 of original volume;
(3), washing: the water extract after concentrating, the absolute ethyl alcohol that adds 4 times while stirring, leave standstill 8～12h, the centrifugal 8～12min of 5000r/min, abandoning supernatant after fully stirring, precipitation is washed 2 times with absolute ethyl alcohol, acetone, ether successively, deionized water redissolution precipitation with minimum volume concentrates freeze-drying again, pulverize, namely get the polysaccharide of entomogenous fungi bond.
Embodiment 1:
Pressing the number of columns ratio gets the raw materials ready
Bacterium liquid 5mL
Several 200 of centipede polypide
To bacterium liquid evenly be sprayed at the polypide surface with watering can, put 18 ℃ then, cultivate 15 days, the beginning blackening appears in polypide after 3 days, mycelia begins the white hypha body after 5 days, is covered with polypide after 7 days, took out polypide in 10 days, natural seasoning namely gets centipede and aweto in conjunction with product.
Embodiment 2:
Pressing the number of columns ratio gets the raw materials ready
Bacterium liquid 5～15mL
Several 200～300 of centipede polypide
Say that above-mentioned bacterium liquid and polypide put into beaker, constantly rock beaker, mix, continue 1 minute, put 18 ℃ then, cultivated 7 days, mycelia begins the white hypha body after 3 days, is covered with polypide after 5 days, took out polypide in 7 days, natural seasoning namely gets centipede and aweto in conjunction with product, carries out polysaccharide determination.
Embodiment 3:
Pressing the number of columns ratio gets the raw materials ready
Bacterium liquid 50～60mL
Several 200～300 of centipede polypide
Say that above-mentioned bacterium liquid and polypide put into beaker, soaked 1 minute, put 20 ℃ then, cultivated 7 days, polypide after 3 days mycelia begin the white hypha body, be covered with polypide after 7 days, take out polypide, natural seasoning obtains the entomogenous fungi bond, carries out polysaccharide determination.
The distinct methods polysaccharide generates content all apparently higher than handling the polypide polyoses content with aweto, and embodiment 3 better effects if by contrast specifically see the following form:
| Classification | Embodiment 1 | Embodiment 2 | Embodiment 3 |
| Polyoses content increases % | 25.17 | 21.24 | 0.63 |
Claims (2)
1. method for preparing the entomogenous fungi bond is characterized in that the preparation method of described entomogenous fungi bond is:
(1), the cultivation of aweto: with aweto toothpick picking, put into the activation of PDY liquid nutrient medium, the preparation spore is collected spore, configuration bacterium liquid;
(2), the preparation of bond: with bacterium liquid and centipede polypide number according to 5～60m1: 200～300 ratio, with watering can bacterium liquid evenly being sprayed at polypide surface or polypide was immersed in the bacterium liquid after 1～3 minute, be placed in 15～25 ℃ the environment, cultivated 7～15 days, the beginning blackening appears in polypide after 3 days, and mycelia begins the white hypha body after 5 days, be covered with polypide after 7 days, took out polypide in 10 days, natural seasoning namely gets centipede and aweto bond; Carry out polysaccharide determination.
2. extracting method of entomogenous fungi bond polysaccharide according to claim 1 is characterized in that this extracting method may further comprise the steps:
(1), dissolving: get the bacterium powder of the entomogenous fungi bond of 50g drying, by the bacterium powder: the mass ratio of water is 1: 25～35 to add deionized water 3L;
(2), extract: after 2h are extracted in 95 ℃ of water-baths, 5000r/min, centrifugal 8～12min gets supernatant, rotary evaporation is concentrated into 1/4 of original volume;
(3), washing: the water extract after concentrating, the absolute ethyl alcohol that adds 4 times while stirring, leave standstill 8～12h, the centrifugal 8～12min of 5000r/min, abandoning supernatant after fully stirring, precipitation is washed 2 times with absolute ethyl alcohol, acetone, ether successively, deionized water redissolution precipitation with minimum volume concentrates freeze-drying again, pulverize, namely get the polysaccharide of polypide bond.
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Citations (5)
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| CN1810842A (en) * | 2005-01-25 | 2006-08-02 | 上海中医药大学 | Longleaf Grateloupia acuminata polysaccharide extract and its prepn and use |
| CN102626418A (en) * | 2012-03-23 | 2012-08-08 | 河南中医学院 | Application of pyrola polysaccharide in preparing immunity-enhancing medicaments and health food |
| CN102653567A (en) * | 2012-04-18 | 2012-09-05 | 刘小龙 | Extracting process of cordyceps sinensis mycelium polysaccharide of composite preparation for preventing tumor |
| CN102688255A (en) * | 2012-06-27 | 2012-09-26 | 江苏省中医药研究院 | Application of polysaccharides extracted from wild chrysanthemum in preparation of medicine for enhancing body immunity |
| CN103070009A (en) * | 2012-12-18 | 2013-05-01 | 徐州鸿宇农业科技有限公司 | Method for cultivating silkworm larvae into silkworm cordyceps by simulating natural ecological environment |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1810842A (en) * | 2005-01-25 | 2006-08-02 | 上海中医药大学 | Longleaf Grateloupia acuminata polysaccharide extract and its prepn and use |
| CN102626418A (en) * | 2012-03-23 | 2012-08-08 | 河南中医学院 | Application of pyrola polysaccharide in preparing immunity-enhancing medicaments and health food |
| CN102653567A (en) * | 2012-04-18 | 2012-09-05 | 刘小龙 | Extracting process of cordyceps sinensis mycelium polysaccharide of composite preparation for preventing tumor |
| CN102688255A (en) * | 2012-06-27 | 2012-09-26 | 江苏省中医药研究院 | Application of polysaccharides extracted from wild chrysanthemum in preparation of medicine for enhancing body immunity |
| CN103070009A (en) * | 2012-12-18 | 2013-05-01 | 徐州鸿宇农业科技有限公司 | Method for cultivating silkworm larvae into silkworm cordyceps by simulating natural ecological environment |
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| 杨永华等: "僵蚕、蜈蚣提取工艺的研究", 《中国中药杂志》, no. 9, 25 September 2001 (2001-09-25) * |
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| 秦葵等: "蝉花的研究概况", 《基层中药杂志》, no. 6, 15 December 2002 (2002-12-15) * |
| 米红霞等: "白僵蚕应用研究进展", 《广东蚕业》, no. 1, 15 March 2010 (2010-03-15) * |
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