CN1456661A - Large scale zhangzhi fungus deep fermenting process - Google Patents
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- CN1456661A CN1456661A CN 03131678 CN03131678A CN1456661A CN 1456661 A CN1456661 A CN 1456661A CN 03131678 CN03131678 CN 03131678 CN 03131678 A CN03131678 A CN 03131678A CN 1456661 A CN1456661 A CN 1456661A
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- 238000000034 method Methods 0.000 title claims description 12
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- 238000004519 manufacturing process Methods 0.000 claims abstract description 15
- 238000000855 fermentation Methods 0.000 claims description 34
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- 238000004458 analytical method Methods 0.000 claims description 10
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- 238000012258 culturing Methods 0.000 claims description 8
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- 239000002054 inoculum Substances 0.000 claims description 4
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- 238000011218 seed culture Methods 0.000 abstract description 4
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
A large-scale deep liquid fermenting process for preparing the mycelia of antrodia cinnamonea includes such steps as collecting the fungus of antrodia cinnamonea, liquid slant culture in shake flask, enlarging the culture, class-one seed culture, fermenting culture, and fermenting in (1000-5000)-L fermentor for 42 hr.
Description
Technical field
The present invention relates to a kind of camphor tree sesame large-scale deep liquid fermentation production technique.
Technical background
Camphor tree sesame (Antrodia camphorata) is a kind of famous Chinese medicine in Taiwan.Wild sporophore growth is at the rotten heartwood inwall of Taiwan protection seeds Cinnamomum kanahirai hay tree trunk, or the surface of the dark humidity of the Cinnamomum kanahirai hay timber of withered lodging, is not easy to be found very much.The camphor tree sesame has long use history in Taiwan, it is most precious medicinal material that the aboriginal all is considered as the camphor tree sesame.Wild top grade under the psychological expectations effect that thing which is rare is dear, also when the river rises the boat goes up day by day for the price of natural camphor tree sesame, the most expensive wild fungus on the world market of being known as at present, market price does not often wait up to more than 10 ten thousand yuan of every jin New Taiwan Currencies to hundreds thousand of units, wildly top gradely reach 15000 dollars/kilogram, the king among the king in the medicinal material of can saying so.At present, can't artificial culture camphor tree sesame sporophore.In Taiwan, it is the protective foods of main raw material that existing in recent years tens companies develop with camphor tree sesame fermentation mycelium and zymophyte powder, can regulate physiological function, nourishing and fit keeping function, physical strength reinforcing, increase immunizing power, spiritual vigorous, enhance metabolism, reduce sense of fatigue.
Modern study finds that the resistance of oxidation of camphor tree sesame tunning is very strong.Taiwan's scholars is investigated in the oxidation resistant ability of external protection normal people red blood corpuscle the Antrodia Camphorata mycelium aqueous extract.By water-soluble peroxidation base disalt acidifying 2, two (the 2-miaow base propane) [2 of 2 '-azo, 2 '-Azobis (2-amidinopropane) dihydrochloride, AAPH] the red blood corpuscle oxidation haemolysis and the lipid/protein peroxidation that are caused suppressed in the mode that a kind of time and concentration rely on by Antrodia Camphorata mycelium.Antrodia Camphorata mycelium also can stop the loss of oxidation resistant gsh (GSH) and Triphosaden (ATP) in the red blood corpuscle kytoplasm.And the infringement of the human endothelial cell that is caused by AAPH also can be suppressed by Antrodia Camphorata mycelium.Interesting is that Antrodia Camphorata mycelium shows the significant cytotoxicity of leukemia HL-60 cell, but human endothelial cell there is not influence.These results show that Antrodia Camphorata mycelium has anti-peroxidation and anticancer composition.The camphor tree sesame substratum dry-matter (DMCM) that Taiwan's scholars is also cultivated deep liquid, the resistance of oxidation, the radical scavenging activity that leach the biologically active substance of different solvents extract in thing (DMF) and the mycelium have been done research.DMF shows the strongest inhibition to lipid peroxidation, and this activity and concentration have functional relation.When 0.2mg/mL, its oxidation resistant activity can be identical with butylated hydroxy anisole (BHA).The resistance of oxidation of mycelium n-hexane extract is minimum, otherwise other mycelium extracts have some to suppress the activity of lipid peroxidation.DMF and mycelium water extract (WEM) show significant free radical scavenging activity.The anti-oxidant activity that leaches thing and mycelium extract is relevant with albumen/polysaccharide ratio in contained polyphenol, total triterpene and the total polysaccharides.Find that in different model researchs the resistance of oxidation of DMCM is lower than DMF, this points out that antioxidant component main among the DMF is mycelial secondary metabolite.Above result shows that DMF can be developed to chemical protector and be used for the treatment of and the free radical diseases associated.
In recent years discover that Crude polysaccharides has the activity of strong anti-hepatitis B virus surface antigen in camphor tree sesame sporophore and the liquid submerged fermentation mycelium, the activity of 50ug/ml polysaccharide concentration hepatitis B virus resisting (HBV) surface antigen is better than the dosage of alpha-interferon 1000U/ml.And the polysaccharide that camphor tree sesame sporophore and some strain liquid submerged fermentation mycelium produce also has the antigenic activity of hepatitis B virus resisting e.This is to find first to derive from that polysaccharide has in the anti-HBV activity of cell levels in the edible fungus.
At present, the artificial culture of camphor tree sesame sporophore not success as yet.The plurality kinds of health care food of selling in Taiwan Province is almost solution fermentation production.China's Mainland relevant camphor tree sesame liquid fermenting patent only has one, and (publication number: CN1309176A), its fermentation is to shake a bottle level by the application of food Industry in Taiwan Institute of Development Studies.Mentioning in Taiwan Province agricultural commission in 2000 the plan of science and technology achievement Report, with 5 liters of fermentor cultivation, is 1.712g/L at the 259.5h mycelial yield.At present, the paper and the patent report that still do not have camphor tree sesame large-scale deep liquid fermentation process aspect.
Summary of the invention
The objective of the invention is to produce Antrodia Camphorata mycelium is target, invents a kind of camphor tree sesame large-scale deep liquid fermentation process of Cheap highly effective, for the large-scale deep liquid fermentation production of camphor tree sesame from now on lays the first stone.
Realize technical scheme of the present invention: with camphor tree sesame 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.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 240-312 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 30-48 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 the shake-flask culture base is housed, 25-33 ℃ of cultivation, rotating speed 120-180rpm/min cultivates after 20-28 hour the access first class seed pot and cultivates, and inoculum size is 4-10%.
(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, keep warm 25-33 ℃ in jar, tank pressure 0.5-0.9kPa, stir speed (S.S.) 80-150rpm/min, air flow 1: 0.3-0.5, fermented 30-48 hour, the fermentor tank of transferring is then cultivated, and inoculum size is 4-10%.
(5) bacterial classification in the first class seed pot is transferred one be equipped with in the fermentor tank of fermention medium, keep jar warm 25-33 ℃, tank pressure 0.5-0.9kPa, stir speed (S.S.) 80-150rpm/min, air flow 1: 0.3-0.5 fermented 35-48 hour.When being reduced to 0.05% left and right sides, reducing sugar content puts jar.
The prescription of slant medium is (g/100ml) among the present invention: potato 15-20, glucose 1-2, peptone 0.1-1, KH
2PO
40.1-0.5, MgSO
40.01-0.1 agar 1.5-2.5 adds water and is settled to volume requiredly, the pH value is 5.0.
The shake-flask culture base is (g/100ml) with shaking bottle prescription of enlarged culturing base among the present invention: glucose 2-4, peptone 0.5-1.5, analysis for soybean powder 0.5-2, wheat bran juice 1.5-2.5, KH
2PO
40.1-0.5, MgSO
40.01-0.1, adding water and be settled to volume requiredly, the pH value is 5.0.
The first order seed culture medium prescription of 100L fermentor tank is (g/100ml) among the present invention: glucose 2-4, peptone 0.5-1.5, analysis for soybean powder 0.5-2, wheat bran juice 0.1-0.5, KH
2PO
40.1-0.5, MgSO
40.05-0.15, adding water and be settled to volume requiredly, the pH value is 5.0.
The fermentative medium formula of 1500L fermentor tank is (g/100ml) among the present invention: glucose 2-6, peptone 0.5-1.5, analysis for soybean powder 0.5-2, wheat bran juice 0.3-0.8, KH
2PO
40.1-0.5, MgSO
40.05-0.15, adding water and be settled to volume requiredly, the pH value is 5.
The shake-flask culture base is 1: 5 with shaking bottle volumetric ratio in the step 2 of the present invention, and the shake-flask culture base is 1: 5 with shaking bottle volumetric ratio in the step 3.
The volume of first class seed pot is 100-300L in the step 4 of the present invention, and the fermention medium 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 0.05%, and microscopy finds that mycelium senesces.
Beneficial effect of the present invention: the present invention is applicable to that large-scale deep liquid fermentation produces the camphor tree sesame, has solved the scale operation of camphor tree sesame and has cultivated a difficult problem of amplifying, and is significant for the development of camphor tree sesame suitability for industrialized production from now on and relevant application industry.The tunning color that the present invention obtains after fermentation ends is bright-coloured rose, gives an attractive smell, and fermented liquid has bitter taste.Dry mycelium is rose.The dry weight of fermentation mycelium is 1-1.5%, and polysaccharide content is 6-8% in the fermentation mycelium.The resistance of oxidation of this fermentation mycelium dry powder is very strong.
Embodiment
The present invention is described in further detail below in conjunction with specific embodiment.
Embodiment 1:
In slant medium, inoculate Antrodia Camphorata mycelium, 28 ℃ of culture temperature, incubation time 264 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 40 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 22 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.) 100 rpm/min, air flow 1: 0.5, 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: 0.5, fermentation time 42 hours.
The slant medium composition is (g/100ml) in the present embodiment 1: potato 15, glucose 2, peptone 0.5, KH
2PO
40.1, MgSO
40.05 agar 2 adds water and is settled to volume requiredly, the pH value is 5.0.
The shake flask fermentation substratum is (g/100ml) with shaking bottle expansion fermention medium composition in the present embodiment 1: glucose 3, peptone 1, analysis for soybean 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.
100L first class seed pot substratum composition is (g/100ml) in the present embodiment 1: glucose 3, peptone 1, analysis for soybean powder 1, wheat bran juice 0.2, 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 5, peptone 1.2, analysis for soybean powder 1.2, wheat bran juice 0.5, KH
2PO
40.25, MgSO
40.125, adding water and be settled to 800L, the pH value is 5.0.
The fermentation ends standard is in the present embodiment: fermented liquid begins the thickness that becomes, and reducing sugar content is reduced to 0.05%, and microscopy finds that mycelium senesces.
Claims (9)
1, a kind of camphor tree sesame liquid submerged fermentation production technique, it is characterized in that: with camphor tree sesame 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.
2, camphor tree sesame liquid submerged fermentation production technique according to claim 1 is characterized in that: being starting strain available from Jinxiang, Shandong fungal studies institute camphor tree sesame fungi.
3, camphor tree sesame liquid submerged fermentation production technique according to claim 1 is characterized in that: it is (g/100ml) that described slant strains substratum is formed: potato 15-20, glucose 1-2, peptone 0.1-1, KH
2PO
40.1-0.5, MgSO
40.01-0.1 agar 1.5-2.5 adds water and is settled to volume requiredly, the pH value is 5.0, and 25-33 ℃ of slant strains culture temperature cultivated 240-312 hour.
4, camphor tree sesame liquid submerged fermentation production technique according to claim 1 is characterized in that: it is (g/100ml) that the substratum of described shake-flask culture is formed: glucose 2-4, peptone 0.5-1.5, analysis for soybean powder 0.5-2, wheat bran juice 1.5-2.5, KH
2PO
40.1-0.5, MgSO
40.01-0.1, adding water and be settled to volume requiredly, the pH value is 5.0, and culture condition is: culture temperature 25-33 ℃, rotating speed 120-180rpm/min cultivated 30-48 hour, and switching is gone into to shake bottle and is carried out enlarged culturing then.
5, camphor tree sesame liquid submerged fermentation production technique according to claim 1 is characterized in that: it is (g/100ml) that the described substratum that shakes bottle enlarged culturing is formed: glucose 2-4, peptone 0.5-1.5, analysis for soybean powder 0.5-2, wheat bran juice 1.5-2.5, KH
2PO
40.1-0.5, MgSO
40.01-0.1, adding water and be settled to volume requiredly, the pH value is 5.0, and culture condition is: culture temperature 25-33 ℃, rotating speed 120-180rpm/min cultivated 20-28 hour, and switching is gone into first order seed and is cultivated then, and inoculum size is 4-10%.
6, camphor tree sesame liquid submerged fermentation production technique according to claim 1 is characterized in that: it is (g/100ml) that the substratum that described first order seed is cultivated is formed: glucose 2-4, peptone 0.5-1.5, analysis for soybean powder 0.5-2, wheat bran juice 0.1-0.5, KH
2PO
40.1-0.5, MgSO
40.05-0.15, adding water and be settled to volume requiredly, the pH value is 5.0, and culture condition is: culture temperature 25-33 ℃, stir speed (S.S.) 80-150rpm/min fermented 30-48 hour, and ferment tank is gone in switching then, and inoculum size is 4-10%.
7, camphor tree sesame liquid submerged fermentation production technique according to claim 1 is characterized in that: described fermentation culture, it is (g/100ml) that fermention medium is formed: glucose 2-6, peptone 0.5-1.5, analysis for soybean powder 0.5-2, wheat bran juice 0.3-0.8, KH
2PO
40.1-0.5, MgSO
40.05-0.15, adding water and be settled to volume requiredly, the pH value is 5.0, culture condition is: culture temperature 25-33 ℃, stir speed (S.S.) 80-150rpm/min fermented 35-48 hour.
8, camphor tree sesame liquid submerged fermentation production technique according to claim 1 is characterized in that: described fermentation culture, and can select the fermentor tank of 1000-5000L to carry out fermentation culture.
9, camphor tree sesame liquid submerged fermentation production technique according to claim 1, it is characterized in that: described fermentation culture, mycelium can reach 1-1.5% with dry weight basis in the fermented liquid.
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| CN 03131678 CN1456661A (en) | 2003-06-03 | 2003-06-03 | Large scale zhangzhi fungus deep fermenting process |
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Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100371450C (en) * | 2005-09-28 | 2008-02-27 | 莱阳农学院 | Fatty acid extract of Antrodiacamphorata mycelium and its uses |
| CN101822170A (en) * | 2010-05-06 | 2010-09-08 | 江南大学 | Method for producing Anrodia camphorata mycelia based on solid-state surface culture |
| CN102047814A (en) * | 2010-10-25 | 2011-05-11 | 青岛农业大学 | Micro ventilation lime wood antrodia camphorate cultivation method |
| CN102172174A (en) * | 2011-03-07 | 2011-09-07 | 江南大学 | Antrodia camphorate quick liquid fermentation process based on asexual spores |
| CN102217487A (en) * | 2011-03-31 | 2011-10-19 | 赵宗杰 | Method for producing mycelia of Antrodia camphorata through deep liquid state fermentation |
| CN102283019A (en) * | 2011-07-06 | 2011-12-21 | 佛山市顺德区今日景艺生物科技有限公司 | Artificial rapid Antrodia camphorata breeding method |
| CN102321684A (en) * | 2011-10-09 | 2012-01-18 | 江南大学 | The biosynthetic fermentation process of a kind of promotion camphor tree sesame active result Antrodin C |
| CN103125270A (en) * | 2013-02-28 | 2013-06-05 | 深圳市仁泰生物科技有限公司 | High-yield antrodia cinnamomea mycelium fermentation method for triterpenoids |
| CN103141891A (en) * | 2013-02-28 | 2013-06-12 | 江南大学 | Preparation method of antrodia camphorata liver protection functional beverage |
| CN103613430A (en) * | 2013-11-25 | 2014-03-05 | 浙江科技学院 | Antrodia cinnamomea growth medium and preparation method and application thereof |
| CN104087631A (en) * | 2014-07-15 | 2014-10-08 | 江苏阜丰生物科技有限公司 | Method for producing Antrodia camphorata extracellular polysaccharides by deep liquid fermentation |
| CN104111302A (en) * | 2014-08-13 | 2014-10-22 | 中山安荞生物科技有限公司 | Method for measuring adenosine content of health-protection antrodia food |
| CN104186880A (en) * | 2014-07-11 | 2014-12-10 | 福建农林大学 | Antrodia mycelium flavor soft sweet and processing method thereof |
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2003
- 2003-06-03 CN CN 03131678 patent/CN1456661A/en active Pending
Cited By (20)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100371450C (en) * | 2005-09-28 | 2008-02-27 | 莱阳农学院 | Fatty acid extract of Antrodiacamphorata mycelium and its uses |
| CN101822170A (en) * | 2010-05-06 | 2010-09-08 | 江南大学 | Method for producing Anrodia camphorata mycelia based on solid-state surface culture |
| CN102047814A (en) * | 2010-10-25 | 2011-05-11 | 青岛农业大学 | Micro ventilation lime wood antrodia camphorate cultivation method |
| CN102172174A (en) * | 2011-03-07 | 2011-09-07 | 江南大学 | Antrodia camphorate quick liquid fermentation process based on asexual spores |
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