CN100567479C - A kind of artificial culturing method of Sanguis Draxonis - Google Patents
A kind of artificial culturing method of Sanguis Draxonis Download PDFInfo
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- CN100567479C CN100567479C CNB2006100107343A CN200610010734A CN100567479C CN 100567479 C CN100567479 C CN 100567479C CN B2006100107343 A CNB2006100107343 A CN B2006100107343A CN 200610010734 A CN200610010734 A CN 200610010734A CN 100567479 C CN100567479 C CN 100567479C
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- sanguis draxonis
- strain
- culturing method
- artificial culturing
- inducible
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- 239000009286 sanguis draxonis Substances 0.000 title claims abstract description 51
- 238000000034 method Methods 0.000 title claims abstract description 15
- 238000012258 culturing Methods 0.000 title claims abstract description 8
- 241000292342 Dracaena cochinchinensis Species 0.000 claims abstract description 16
- 230000001939 inductive effect Effects 0.000 claims abstract description 16
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims abstract description 13
- 239000009136 dragon's blood Substances 0.000 claims abstract description 12
- 235000015097 nutrients Nutrition 0.000 claims abstract description 9
- 238000000926 separation method Methods 0.000 claims abstract description 7
- 230000001954 sterilising effect Effects 0.000 claims abstract description 5
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 4
- 238000000605 extraction Methods 0.000 claims abstract description 3
- 240000004824 Trimezia steyermarkii Species 0.000 claims description 10
- 230000004913 activation Effects 0.000 claims description 10
- 239000012153 distilled water Substances 0.000 claims description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 8
- 229920001817 Agar Polymers 0.000 claims description 7
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 7
- 244000061456 Solanum tuberosum Species 0.000 claims description 7
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 7
- 239000008272 agar Substances 0.000 claims description 7
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims description 7
- 239000008103 glucose Substances 0.000 claims description 7
- 241000233866 Fungi Species 0.000 claims description 6
- 239000001965 potato dextrose agar Substances 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 6
- 238000004321 preservation Methods 0.000 claims description 6
- 241000223194 Fusarium culmorum Species 0.000 claims description 4
- 239000002609 medium Substances 0.000 claims description 4
- 239000000284 extract Substances 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- 238000011534 incubation Methods 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 claims description 2
- 238000005070 sampling Methods 0.000 claims description 2
- 241001043298 Croton draco Species 0.000 abstract 1
- 244000005700 microbiome Species 0.000 description 6
- 238000004519 manufacturing process Methods 0.000 description 5
- 239000008280 blood Substances 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- 238000004128 high performance liquid chromatography Methods 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 238000000855 fermentation Methods 0.000 description 3
- 230000004151 fermentation Effects 0.000 description 3
- 230000007812 deficiency Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- LZLCKUZWICVGNZ-UHFFFAOYSA-N 1-(4-hydroxyphenyl)-3-(2,4,6-trimethoxycyclohexa-2,4-dien-1-yl)prop-2-en-1-one Chemical compound COC1C=C(OC)C=C(OC)C1C=CC(=O)C1=CC=C(O)C=C1 LZLCKUZWICVGNZ-UHFFFAOYSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000590428 Panacea Species 0.000 description 1
- 208000025865 Ulcer Diseases 0.000 description 1
- 244000112726 Ximenia americana Species 0.000 description 1
- 235000006801 Ximenia americana Nutrition 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- RCTYPNKXASFOBE-UHFFFAOYSA-M chloromercury Chemical compound [Hg]Cl RCTYPNKXASFOBE-UHFFFAOYSA-M 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 230000035876 healing Effects 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 229960002523 mercuric chloride Drugs 0.000 description 1
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000005477 standard model Effects 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 230000031068 symbiosis, encompassing mutualism through parasitism Effects 0.000 description 1
- 230000017423 tissue regeneration Effects 0.000 description 1
- 231100000397 ulcer Toxicity 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a kind of artificial culturing method of Sanguis Draxonis.The position that is forming dragon's blood in the strain of wild Dracaena cochinchinensis live body stem is cut off, obtain the Sanguis Draxonis inducible strain through sterilization, cultivation, separation with after identifying, in the nutrient solution of every 1000ml, put into the mycelium of 0.5～1.5g, shaking culture is after 7～28 days under unglazed photograph, 25～27 ℃ of constant temperature and 220～280r/min, use n-butanol extraction, after separating propyl carbinol, what obtain is Sanguis Draxonis.The present invention has opened up the new way that Sanguis Draxonis is produced, and its method is easy, with low cost, has better market prospect.
Description
Technical field
The invention belongs to biological technical field, be specifically related to a kind of method of utilizing fungi fermentation technology artificial culture Sanguis Draxonis.
Background technology
Dragon's blood is rare rare Chinese medicine, has promoting blood circulation and removing blood stasis, swelling and pain relieving, astringing to arrest bleeding, and promoting tissue regeneration and ulcer healing, the effect of enriching blood are usually used in various mass formed by blood stasis, are called " with the panacea of blood " (seeing Compendium of Material Medica).Dragon's blood derives from the Western Regions at first, the Su Mendala that modern age is inferior southeast.The seventies, the Cai Xitao professor of Xishuangbanna Tropical Plant Garden, Chinese Academy of Sciences has found to produce plant resources---the Dracaena cochinchinensis (Dracaena cochinchinensis) of dragon's blood in south of Yunnan, and, develop domestic Dragon Blood by vegetable chemistry and pharmaceutical research, be called Sanguis Draxonis.Experimental results show that the dragon's blood that is come by the Western Regions put down in writing in homemade Sanguis Draxonis and the Compendium of Material Medica (dragon ' s blood) is the most approaching, can be used as the homemade alternative articles for use of South East Asia import dragon's blood fully.Family surplus the existing Sanguis Draxonis manufacturing enterprise ten of China contains the dragon's blood medicine kind more than 60 nearly that tallow wood is a raw material production with Dracaena cochinchinensis, and annual value of production reaches more than 500,000,000 yuan.
Under state of nature, after Dracaena cochinchinensis damaged because of thunder and lightning, storm and phytophagy animal gnaw, the defensive compound owing to the intrusion of microorganism produces was Sanguis Draxonis.But this process is very slow, and often decades-long and even upward century-old is difficult to satisfy the growing market requirement.At present, to Dracaena cochinchinensis symbiosis fungi and with relation that Sanguis Draxonis forms a spot of preliminary discussion is only arranged, though the hypothesis that has proposed to utilize the fungal induction Sanguis Draxonis to produce, not seeing as yet in the prior art has Sanguis Draxonis to induce the separation and Culture of microorganism and utilizes the fungi fermentation technology to produce the report of Sanguis Draxonis at indoor culture.
Summary of the invention
The objective of the invention is at the deficiencies in the prior art, a kind of method of artificial culture Sanguis Draxonis is provided, it is numerous to utilize the Sanguis Draxonis inducible strain manually to expand, and realizes the suitability for industrialized production of Sanguis Draxonis.
The invention provides a kind of artificial culturing method of Sanguis Draxonis, this method is made up of following steps:
1, Sanguis Draxonis is induced sampling and the separation and Culture of fungi: the position that is forming dragon's blood in the strain of wild Dracaena cochinchinensis live body stem is cut off, obtain the Sanguis Draxonis inducible strain through sterilization, cultivation, separation with after identifying, and standby 4 ℃ of following preservations; This bacterial strain now is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC), formal name used at school is accredited as Fusarium culmorum, bacterial strain called after Sanguis Draxonis inducible strain Brwg, deposit number is CGMCC No 1621, preservation date on February 20th, 2006.
2, the preparation of strain cultured solution: with the fresh stem of Dracaena cochinchinensis air-dry after, it is standby to be ground into 60～100 purpose powder.Make strain cultured solution after organizing powder to stir 100 parts of distilled water, 20 parts of potato juices, 2 parts of glucose, 0.2 part of agar and 0.5 part of Dracaena cochinchinensis stem, it is standby to sterilize;
3. the activation of inducible strain: place activation medium to make its activation the Sanguis Draxonis inducible strain;
4, firm activatory Sanguis Draxonis inducible strain is added in the described strain cultured solution cultivate, put into the mycelium of 0.5～1.5g in every 1000ml nutrient solution, shaking culture is 7～21 days under unglazed photograph, 25 ℃～27 ℃ constant temperature and 280～320r/min;
5, the extraction of Sanguis Draxonis composition in the nutrient solution: the nutrient solution after will fermenting is poured separating funnel into, adds the propyl carbinol of same volume, behind the mixing that fully vibrates, extracts repeatedly 3 times, isolates the propyl carbinol part, and propyl carbinol is distilled, and what obtain is Sanguis Draxonis.
The described activation medium of step 3 is a potato dextrose agar.
The weight part proportioning of described potato dextrose agar is: potato juice 20, glucose 2, agar 1.5 and distilled water 76.5.
The described incubation time the best of step 4 is 14～21 days.
The present invention has overcome the deficiencies in the prior art, taken the lead in realizing that Sanguis Draxonis induces the separation and Culture of microorganism, and then it is numerous to utilize these microorganisms manually to expand, and produces Sanguis Draxonis at indoor culture, has realized the suitability for industrialized production of Sanguis Draxonis.The inventive method is easy, with low cost, is implemented as the rarity that utilizes better in this traditional medicine of Sanguis Draxonis and has created advantageous conditions, has better market prospect.The explanation of preservation biomaterial
The Sanguis Draxonis inducible strain that the present invention relates to now is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC), formal name used at school is accredited as Fusarium culmorum, bacterial strain called after Sanguis Draxonis inducible strain Brwg, deposit number is CGMCC No 1621, preservation date on February 20th, 2006.
Description of drawings
Fig. 1 is used for the molecular formula of the standard model (4 '-hydroxyl-2,4,6-trimethoxy dihydrochalcone) of Sanguis Draxonis quality inspection for Nat'l Pharmaceutical ﹠ Biological Products Control Institute;
Situation when Fig. 2 cultivates Sanguis Draxonis for the present invention.
Embodiment
Below in conjunction with embodiment the present invention is described in further detail, but they are not limitation of the invention.
Embodiment 1
The position that is forming dragon's blood in the strain of wild Dracaena cochinchinensis live body stem is cut off, with 0.1% mercuric chloride (HgCl
2) carry out 5 minutes surface sterilization and 6 distilled water rinsings; After Dracaena cochinchinensis tissue after will sterilizing in the Bechtop is cut into 0.8 * 0.5 * 0.3 centimetre pane, puts in the potato dextrose agar (all the other are distilled water for the juice of 20% potato, 2% glucose and 1.5% agar) and cultivate; After separating, identifying fungi, obtain Sanguis Draxonis inducible strain Brwg (Fusarium culmorum), standby 4 ℃ of following preservations.With the fresh stem of Dracaena cochinchinensis air-dry after, it is standby to be ground into 80 purpose powder.Get and make strain cultured solution after 100 parts of distilled water, 20 parts of potato juices, 2 parts of glucose, 0.2 part of agar and 0.5 part of Dracaena cochinchinensis stem organize powder to stir, it is standby to sterilize.The Sanguis Draxonis inducible strain placed in the potato dextrose agar (all the other are distilled water for the juice of 20% potato, 2% glucose and 1.5% agar) make its activation.Sanguis Draxonis inducible strain after the activation is added in the described strain cultured solution and cultivates, put into the mycelium of 1g in every 1000ml nutrient solution, shaking culture is 7 days under unglazed photograph, 26 ℃ of constant temperature and 250r/min; Pour the nutrient solution after the fermentation into separating funnel, add the propyl carbinol of same volume, behind the mixing that fully vibrates, extract repeatedly 3 times, isolate n-butanol portion, propyl carbinol is distilled, what obtain is Sanguis Draxonis, detects through high performance liquid chromatography, and its lourerin content reaches 0.011～0.013%.
Embodiment 2
Repeat embodiment 1, following difference is arranged: shaking culture 14 days, the Sanguis Draxonis that obtains detects through high performance liquid chromatography, and its lourerin content reaches 0.121～0.142%.
Embodiment 3
Repeat embodiment 1, following difference is arranged: shaking culture 17 days, the Sanguis Draxonis that obtains detects through high performance liquid chromatography, and its lourerin content reaches 0.192～0.212%.
Embodiment 4
Repeat embodiment 1, following difference is arranged: shaking culture 21 days, the Sanguis Draxonis that obtains detects through high performance liquid chromatography, and its lourerin content reaches 0.378～0.391%.
Claims (4)
1, a kind of artificial culturing method of Sanguis Draxonis, this method is made up of following steps:
(1) Sanguis Draxonis is induced sampling and the separation and Culture of fungi: the position that is forming dragon's blood in the strain of wild Dracaena cochinchinensis live body stem is cut off, obtain Sanguis Draxonis inducible strain (Fusarium culmorum) through sterilization, cultivation, separation with after identifying, its deposit number is CGMCC No 1621, and standby 4 ℃ of following preservations;
(2) preparation of strain cultured solution: with the fresh stem of Dracaena cochinchinensis air-dry after, it is standby to be ground into 60～100 purpose powder; Make strain cultured solution after organizing powder to stir 100 parts of distilled water, 20 parts of potato juices, 2 parts of glucose, 0.2 part of agar and 0.5 part of Dracaena cochinchinensis stem, it is standby to sterilize;
(3) activation of inducible strain: place activation medium to make its activation the Sanguis Draxonis inducible strain;
(4) firm activatory Sanguis Draxonis inducible strain is added in the described strain cultured solution cultivate, put into the mycelium of 0.5～1.5g in every 1000ml nutrient solution, shaking culture is 7～21 days under unglazed photograph, 25～27 ℃ of constant temperature and 220～280r/min;
(5) extraction of Sanguis Draxonis composition in the nutrient solution: the nutrient solution after will fermenting is poured separating funnel into, adds the propyl carbinol of same volume, behind the mixing that fully vibrates, extracts repeatedly 3 times, isolates the propyl carbinol part, and propyl carbinol is distilled, and what obtain is Sanguis Draxonis.
2, the artificial culturing method of Sanguis Draxonis according to claim 1 is characterized in that: the described activation medium of step 3 is a potato dextrose agar.
3, the artificial culturing method of Sanguis Draxonis according to claim 2 is characterized in that: the weight part proportioning of described potato dextrose agar is: potato juice 20, glucose 2, agar 1.5 and distilled water 76.5.
4, the artificial culturing method of Sanguis Draxonis according to claim 1 is characterized in that: the described incubation time of step 4 is 14～21 days.
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| CNB2006100107343A CN100567479C (en) | 2006-03-09 | 2006-03-09 | A kind of artificial culturing method of Sanguis Draxonis |
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| CNB2006100107343A CN100567479C (en) | 2006-03-09 | 2006-03-09 | A kind of artificial culturing method of Sanguis Draxonis |
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| CN100567479C true CN100567479C (en) | 2009-12-09 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI812448B (en) * | 2022-09-02 | 2023-08-11 | 吳俊毅 | Hydroxyloureirin, production method and use thereof |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN100531559C (en) * | 2007-04-12 | 2009-08-26 | 中国热带农业科学院热带生物技术研究所 | Method of artificial cultivation for dragon trees to generate dragon's blood |
| CN102649938B (en) * | 2011-02-24 | 2013-05-29 | 中国医学科学院药用植物研究所 | Two fungi for inducing dragon tree to produce dragon's blood |
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2006
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Non-Patent Citations (2)
| Title |
|---|
| 血竭资源开发的研究进展. 陈定方等.广州中医药大学学报，第21卷第6期. 2004 |
| 血竭资源开发的研究进展. 陈定方等.广州中医药大学学报，第21卷第6期. 2004 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI812448B (en) * | 2022-09-02 | 2023-08-11 | 吳俊毅 | Hydroxyloureirin, production method and use thereof |
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| CN1821388A (en) | 2006-08-23 |
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