CN1281734C - Technology for producing birch brown fungus extract and saccharide using large scale liquid submerged fermentation process - Google Patents
Technology for producing birch brown fungus extract and saccharide using large scale liquid submerged fermentation process Download PDFInfo
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- CN1281734C CN1281734C CN 200410013868 CN200410013868A CN1281734C CN 1281734 C CN1281734 C CN 1281734C CN 200410013868 CN200410013868 CN 200410013868 CN 200410013868 A CN200410013868 A CN 200410013868A CN 1281734 C CN1281734 C CN 1281734C
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Abstract
The present invention relates to a technology for producing an aqueous extract of inonotus obliquus and polysaccharide of the inonotus obliquus by using liquid submerged fermentation on a large scale. The present invention aims to produce the aqueous extract of the inonotus obliquus and the polysaccharide of the inonotus obliquus by using the liquid submerged fermentation on a large scale. Inonotus obliquus from the fungus academy in Jinxiang of Shandong province is used as an original strain, the slope strain is treated by liquid culture in a shake flask, liquid enlarging culture in the shake flask, first-level seed culture and fermentation culture, and the final volume of a fermentation tank reaches 1000 to 5000L. After the fermentation is finished, the temperature of the final fermentation liquid is heated to 60 to 100 DEG C, and the final fermentation liquid is stirred for 2 to 4 hours at a stirring speed of 150 to 250 rpm/min. The fermentation liquid is delivered into a storing tank by a pump, and then is treated by sheet frame press filtration, clear liquid concentration and spray drying to obtain the aqueous extract of the inonotus obliquus. If the clear liquid is treated by a proper amount of ethanol for precipitation after being concentrated, the precipitation can be dissolved by adding a proper amount of water, and then the polysaccharide of the inonotus obliquus can be obtained by spray drying.
Description
Technical field
The present invention relates to a kind of large-scale deep liquid fermentation and produce Phaeopoms obliquus water extract and polysaccharide process.
Technical background
Phaeopoms obliquus (Fuscoporia obliqua) has another name called Chaga, at Russia and the Eastern Europe Chaga that cries among the people, belong to Basidiomycotina, Hymenomycetes, non-brown Zoopagales, polyporaceae, brown transverse hole fungus and belong to, the bark of being born in white birch, silvery birch, elm, alder etc. down or the bark of the standing tree of living down or on felling back trees dried-up.
16-17 is since century, and Russia, Poland, Finland etc. are among the people just extensively to utilize its wild sporophore to prevent and treat various difficult and complicated cases, as various cancers (various digestion organs cancers such as cancer of the stomach, liver cancer, intestinal cancer) and heart trouble and diabetes etc.Poland just has been extensive use of Phaeopoms obliquus and treats cancer since 1961.The researchist of Japan speaks highly of this medicine among the people of Russia, and thinks that this is a kind of peculiar present that God grants the suffering mankind, can be used to prevent and treat liver cancer, AIDS and O-157 intestinal bacteria and poisons.It is reported that the smart powder of the Phaeopoms obliquus that the Muscovite Suo Molesiji of bearing (Komsomlski) drugmaker produces reaches 93% to the curative ratio of diabetes.The decocting Stewed Dish of Phaeopoms obliquus has notable antitumor activity.The syrup of Phaeopoms obliquus is used for the treatment of cancer already.Because the source is rare, the price of its wild sporophore is 7-8 a times of Phellinus (Phellinus igniarius).
Summary of the invention
The objective of the invention is to produce Phaeopoms obliquus water extract and polysaccharide is target, invents a kind of Phaeopoms obliquus water extract of Cheap highly effective and the large-scale deep liquid fermentation process of polysaccharide, for from now on commercial application lays the first stone.
Realize technical scheme of the present invention: with the Phaeopoms obliquus fungi is starting strain, adopts slant strains to carry out liquid shaking bottle cultivation, liquid shaking bottle enlarged culturing, first order seed cultivation and fermentation culture.After fermentation ends, the end of a period fermented liquid is continued to be heated to certain temperature, stir and keep certain hour, after stirring stops, fermented liquid spray drying is got zymophyte powder.As with fermented liquid through filter press, clear liquid concentrates, spraying drying obtains the Phaeopoms obliquus water extract, use an amount of ethanol sedimentation, taking precipitate to add suitable quantity of water to dissolve as clear liquid being concentrated the back, spraying drying can get Fuscoporia obliqua polysaccharide.
This bacterial strain is available from Jinxiang, Shandong fungal studies institute.
Realize that concrete steps of the present invention are as follows:
(1) takes out slant strains and insert in the fresh slant medium of preparing, culture temperature 25-33 ℃, cultivated 150-250 hour.
(2) slant strains in the step 1 is transferred the shaking in the bottle of shake-flask culture base is housed, 25-33 ℃ of cultivation, rotating speed 120-180rpm/min cultivates the enlarged culturing of shaking bottle after 70-120 hour.
(3) bottle bacterial classification that shakes in the step 2 is transferred and carried out enlarged culturing again into the bottle that shakes that shakes bottle enlarged culturing base is housed, 25-33 ℃ of cultivation, rotating speed 120-180rpm/min cultivates after 35-60 hour the access first class seed pot and cultivates.
(4) bottle bacterial classification that shakes in the step 3 is transferred one and is equipped with in the first class seed pot of seed culture medium, inoculum size is 4-10%, keep warm 25-33 ℃ in jar, tank pressure 0.5-0.9kPa, stir speed (S.S.) 80-150rpm/min, air flow 1: 0.3-0.5v/v.m fermented 35-60 hour, and ferment tank is gone in switching then.
(5) bacterial classification in the first class seed pot is transferred one be equipped with in the fermentor tank of fermention medium, inoculum size is 4-10%, keeps jar warm 25-33 ℃, tank pressure 0.5-0.9kPa, stir speed (S.S.) 80-150rpm/min, air flow 1: 0.3-0.5v/v.m fermented 90-150 hour.When being reduced to the 1.0-2.0% left and right sides, the fermented liquid reducing sugar content, puts jar for fermentation ends.
The prescription of slant medium is (g/100ml) among the present invention: potato 15-25, glucose 1-3, agar 1.5-2.5, add water be settled to volume required, pH value nature.
The prescription of shake-flask culture base is (g/100ml) among the present invention: glucose 2-4, peptone 0.05-0.5, wheat bran juice 1.0-2.5, KH
2PO
40.1-0.5, MgSO
40.01-0.2, adding water and be settled to volume requiredly, the pH value is 4.5-5.5.
Shaking bottle prescription of enlarged culturing base among the present invention is (g/100ml): glucose 2-4, peptone 0.05-0.7, Semen Maydis powder 0.5-2, wheat bran juice 1.0-2.5, KH
2PO
40.1-0.5, MgSO
40.01-0.2, adding water and be settled to volume requiredly, the pH value is 4.5-5.5.
The culture medium prescription of first order seed cultivation and fermentation culture is (g/100ml) among the present invention: glucose 2-4, peptone 0.05-0.7, Semen Maydis powder 0.5-2, analysis for soybean powder 0.5-2, wheat bran juice 1-3, KH
2PO
40.1-0.5, MgSO
40.01-0.2, adding water and be settled to volume requiredly, the pH value is 4.5-5.5.
The shake-flask culture base is 1: 5 with shaking bottle volumetric ratio in the step 2 of the present invention and 3.
The volume of first class seed pot is 100-300L in the step 4 of the present invention, and the first order seed substratum that is equipped with in this first class seed pot should be 50-180L mutually; The volume of fermentor tank is 1000-5000L in the step 5, and the fermention medium that is equipped with in this fermentor tank should be 500-3500L mutually.
Jar standard of putting of the present invention is that fermented liquid begins the thickness that becomes, and reducing sugar content is reduced to about 1.0-2.0%, and microscopy finds that mycelium senesces.
Obtain its water extract and polysaccharide as need, then after fermentation of the present invention stopped, the end of a period liquid temp that will ferment was heated to 60-100 ℃, and stirring velocity is 150-250rpm/mm, kept 2-4 hour, after stirring stops, fermented liquid spray drying was got zymophyte powder.Obtain the Phaeopoms obliquus water extract as need, fermented liquid is transported in the basin,, get clear liquid through filter press with pump.Concentrating filter liquor, making last volume is the 1/4-1/10 of stoste, gets the concentrated solution spraying drying and obtains the Phaeopoms obliquus water extract.As add an amount of ethanol and make alcohol concn reach 75%, alcohol was analysed 20-30 hour, got the precipitate with deionized water dissolving, and spraying drying promptly obtains Fuscoporia obliqua polysaccharide.
Beneficial effect of the present invention: the present invention is applicable to that large-scale deep liquid fermentation produces Phaeopoms obliquus water extract and polysaccharide, and is significant for the development of from now on suitability for industrialized production and relevant application industry.Gained Phaeopoms obliquus water extract of the present invention and polysaccharide color are brown.
Embodiment
The present invention is described in further detail below in conjunction with specific embodiment.
Embodiment 1:
In slant medium the inoculation available from Jinxiang, Shandong fungal studies the Phaeopoms obliquus mycelium, 28 ℃ of culture temperature, incubation time 180 hours.Fresh slant strains access is equipped with in the 500ml triangular flask of 100ml substratum, and 28 ℃ of shaking tables were cultivated shaking speed 150rpm/min 96 hours.Then gained is shaken transfer 500ml triangle that the 100ml substratum is housed of bottle bacterial classification and shake and carry out enlarged culturing in the bottle, 28 ℃ of shaking tables were cultivated shaking speed 150rpm/min 45 hours.During the bacterial classification 2200ml that triangular flask is enlarged transfers the 100L first class seed pot that the 50L seed culture medium is housed then, keep 28 ℃ of jar temperature, tank pressure 0.7kPa, stir speed (S.S.) 100rpm/min, air flow 1: 0.5v/v.m, fermentation time 45 hours.The first class seed pot bacterial classification is transferred in the 1500L fermentor tank that the 800L fermention medium is housed, keep 28 ℃ of jar temperature, tank pressure 0.7kPa, stir speed (S.S.) 100rpm/min, air flow 1: 0.5v/v.m, fermentation time 120 hours, zymophyte powder counts 3.5% with dry weight, and polysaccharide content is 22%.
To form be (g/100ml) to slant medium in the present embodiment 1: potato 20, and glucose 2, agar 2 adds water and is settled to volume requiredly, and the pH value is a nature.
Shake-flask culture base composition is (g/100ml) in the present embodiment 1: glucose 3, peptone 0.2, wheat bran juice 2, KH
2PO
40.2, MgSO
40.1, adding water and be settled to volume requiredly, the pH value is 5.0.
Shaking a bottle enlarged culturing base composition in the present embodiment 1 is (g/100ml): glucose 3, peptone 0.2, Semen Maydis powder 1, wheat bran juice 2, KH
2PO
40.2, MgSO
40.1, adding water and be settled to volume requiredly, the pH value is 5.0.
The substratum of 100L first class seed pot composition is (g/100ml) in the present embodiment 1: glucose 3, peptone 0.2, Semen Maydis powder 1.5, analysis for soybean powder 1, wheat bran juice 2, KH
2PO
40.2, MgSO
40.1, adding water and be settled to 50L, the pH value is 5.0.
The culture medium prescription of 1500L fermentor tank is (g/100ml) in the present embodiment 1: glucose 3, peptone 0.5, Semen Maydis powder 1.5, analysis for soybean powder 1.0, wheat bran juice 2, KH
2PO
40.2, MgSO
40.1, adding water and be settled to 800L, the pH value is 5.0.
The fermentation ends standard is in the present embodiment 1: fermented liquid begins the thickness that becomes, and reducing sugar content is reduced to 1.5%, and microscopy finds that mycelium senesces.
The method of obtaining Phaeopoms obliquus zymophyte powder, water extract and polysaccharide is: after fermentation of the present invention stops, temperature is heated to 90 ℃, stirring velocity is 220rpm/min, keeps 3 hours, after stirring stops, fermented liquid spray drying is got zymophyte powder.Or with pump fermented liquid is transported in the basin, through filter press, concentrating filter liquor, making last volume is 1/5 of stoste, gets the concentrated solution spraying drying and obtains the Phaeopoms obliquus water extract.Obtain polysaccharide as need, then concentrated solution is added an amount of ethanol and make alcohol concn reach 75%, alcohol was analysed 24 hours, got the precipitate with deionized water dissolving, and spraying drying obtains Fuscoporia obliqua polysaccharide.
Claims (3)
1, a kind of liquid submerged fermentation is produced the Phaeopoms obliquus zymophyte powder, the technology of water extract and polysaccharide, it is characterized in that with Phaeopoms obliquus fungi (Fuscoporia obliqua) be starting strain, adopting slant strains to carry out liquid shaking bottle cultivates, the liquid shaking bottle enlarged culturing, first order seed is cultivated and fermentation culture gets the Phaeopoms obliquus mycelium fermentation broth, after fermentation ends, the end of a period fermented liquid is continued to be heated to certain temperature, certain hour is kept in stirring, after stirring stops fermented liquid spray drying being got zymophyte powder, fermented liquid is through filter press, clear liquid concentrates, spraying drying obtains the Phaeopoms obliquus water extract, or with an amount of ethanol sedimentation in the concentrated back of clear liquid, taking precipitate adds the suitable quantity of water dissolving, and spraying drying can get Fuscoporia obliqua polysaccharide;
Slant strains is cultivated, and it is in g/100ml that substratum is formed: potato 15-25, and glucose 1-3, agar 1.5-2.5 adds water and is settled to volume requiredly, and the pH value is a nature, and 25-33 ℃ of slant strains culture temperature cultivated 150-250 hour;
Shake-flask culture, it is in g/100ml that substratum is formed: glucose 2-4, peptone 0.05-0.5, wheat bran juice 1.0-2.5, KH
2PO
40.1-0.5, MgSO
40.01-0.2, adding water and be settled to volume requiredly, the pH value is 4.5-5.5, and culture condition is: culture temperature 25-33 ℃, rotating speed 120-180rpm/min cultivated 70-120 hour, and switching is gone into to shake bottle and is carried out enlarged culturing then;
Shake a bottle enlarged culturing, it is in g/100ml that substratum is formed: glucose 2-4, peptone 0.05-0.7, Semen Maydis powder 0.5-2, wheat bran juice 1.0-2.5, KH
2PO
40.1-0.5, MgSO
40.01-0.2, adding water and be settled to volume requiredly, the pH value is 4.5-5.5, and culture condition is: culture temperature 25-33 ℃, rotating speed 120-180rpm/min cultivated 35-60 hour, and switching is gone into first class seed pot and is cultivated then;
First order seed is cultivated, and it is in g/100ml that substratum is formed: glucose 2-4, peptone 0.05-0.7, Semen Maydis powder 0.5-2, analysis for soybean powder 0.5-2, wheat bran juice 1-3, KH
2PO
40.1-0.5, MgSO
40.01-0.2, adding water is settled to volume required, the pH value is 4.5-5.5, inoculum size is 4%-10%, and culture condition is: culture temperature 25-33 ℃, and tank pressure 0.5-0.9kPa, stir speed (S.S.) 80-150rpm/min, air flow 1: 0.3-0.5v/v.m fermented 35-60 hour, and ferment tank is gone in switching then;
Fermentation culture, it is in g/100ml that fermention medium is formed: glucose 2-4, peptone 0.05-0.7, Semen Maydis powder 0.5-2, analysis for soybean powder 0.5-2, wheat bran juice 1-3, KH
2PO
40.1-0.5, MgSO
40.01-0.2, adding water is settled to volume required, the pH value is 4.5-5.5, and inoculum size is 4%-10%, and culture condition is: culture temperature 25-33 ℃, tank pressure 0.5-0.9kPa, stir speed (S.S.) 80-150rpm/min, air flow 1: 0.3-0.5v/v.m fermented 90-150 hour, when the fermented liquid reducing sugar content drops to 1.0%-2.0%, for fermentation ends;
End of a period fermented liquid temperature is heated to 60-100 ℃, and stirring velocity is 150-250rpm/min, keeps 2-4 hour, and stirring can get the Phaeopoms obliquus zymophyte powder with fermented liquid with spraying drying after finishing; Or with pump fermented liquid is transported in the basin, obtaining clear liquid through filter press, clear liquid is concentrated into the 1/4-1/10 that volume is an original volume, and spraying drying obtains the Phaeopoms obliquus water extract; Or clear liquid is concentrated the back, and use an amount of ethanol sedimentation, alcohol to analyse the liquid alcohol concn be 75%, and alcohol was analysed 20-30 hour, and taking precipitate adds suitable quantity of water and dissolves, and spraying drying can get Fuscoporia obliqua polysaccharide.
2, liquid submerged fermentation according to claim 1 is produced the technology of Phaeopoms obliquus zymophyte powder, water extract and polysaccharide, it is characterized in that with available from Jinxiang, Shandong fungal studies the Phaeopoms obliquus fungi be starting strain.
3, liquid submerged fermentation according to claim 1 is produced the technology of Phaeopoms obliquus zymophyte powder, water extract and polysaccharide, it is characterized in that described fermentation culture, selects the fermentor tank of 1000-5000L to carry out fermentation culture.
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|---|---|---|---|
| CN 200410013868 CN1281734C (en) | 2004-01-07 | 2004-01-07 | Technology for producing birch brown fungus extract and saccharide using large scale liquid submerged fermentation process |
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| CN 200410013868 CN1281734C (en) | 2004-01-07 | 2004-01-07 | Technology for producing birch brown fungus extract and saccharide using large scale liquid submerged fermentation process |
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| CN1281734C true CN1281734C (en) | 2006-10-25 |
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Families Citing this family (13)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101748159B (en) * | 2008-12-02 | 2012-09-05 | 徐州师范大学 | Method for culturing inonotus obliquus by fermentation |
| CN102108371A (en) * | 2009-12-23 | 2011-06-29 | 徐向群 | Process for submerged fermentation and production of anti-oxidation active substances of white rot fungi by utilizing lignocellulosic agricultural and forestry wastes |
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| CN103833863A (en) * | 2012-11-20 | 2014-06-04 | 中国科学院兰州化学物理研究所 | Technology for preparing crude polysaccharides by extracting polysaccharides from Enteromorpha prolifera |
| CN103156052B (en) * | 2013-02-21 | 2014-06-04 | 徐向群 | Submerged fermentation technology of producing immunological enhancement active materials of inonotus obliquus |
| CN104956925B (en) * | 2015-07-24 | 2018-01-30 | 刘随记 | The production method of the continuous submerged fermentation liquid of Inonotus obliquus and yeast powder |
| CN105349439A (en) * | 2015-12-16 | 2016-02-24 | 黑龙江众生生物工程有限公司 | Culture and fermentation method of inonotus obliquus and preparation method of water-soluble polysaccharides of inonotus obliquus |
| CN105794955A (en) * | 2016-03-21 | 2016-07-27 | 东北农业大学 | Inonotus obliquus selenizing polysaccharide preparation and application of inonotus obliquus selenizing polysaccharide preparation for fresh keeping of raspberries |
| CN106579293A (en) * | 2016-11-22 | 2017-04-26 | 汪逸凡 | Preparation method of digestion-promoting compound feed sweetener |
| CN107916229B (en) * | 2017-10-23 | 2019-11-08 | 山东省农业科学院畜牧兽医研究所 | One plant of Inonotus obliquus and its application |
| KR102195870B1 (en) * | 2017-10-23 | 2020-12-28 | 인스티튜트 오브 애니멀 사이언스 앤드 베테리너리 메디컬 산동 아카데미 오브 어그리컬쳐 사이언스 | Chaga fungus and its application |
| CN109965268A (en) * | 2019-04-12 | 2019-07-05 | 天津农学院 | A kind of edible fungus jelly and preparation method thereof |
| CN110734932B (en) * | 2019-07-29 | 2021-10-22 | 浙江养生堂天然药物研究所有限公司 | Fermented birch juice and its production method |
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