CN1183138C - Process for promoting generation of 10-hydroxycamptothecin in the bud of camptotheca seed - Google Patents
Process for promoting generation of 10-hydroxycamptothecin in the bud of camptotheca seed Download PDFInfo
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- CN1183138C CN1183138C CNB021329222A CN02132922A CN1183138C CN 1183138 C CN1183138 C CN 1183138C CN B021329222 A CNB021329222 A CN B021329222A CN 02132922 A CN02132922 A CN 02132922A CN 1183138 C CN1183138 C CN 1183138C
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- hydroxycamptothecine
- camptotheca
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- hydroxycamptothecin
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Abstract
The present invention relates to a method for promoting the generation of 10-hydroxycamptothecin in camptotheca acuminata seed sprouts, which comprises the following technological processes: camptotheca acuminata seeds are cultured by water, and the camptotheca acuminata seeds are germinated; the germinated camptotheca acuminata seeds are put in an ultrasonic device filled with water for ultrasonic radiation; then the radiated camptotheca acuminata seed sprouts are put in a homogenizer filled with ethanol for seed sprout tissue pulverization, homogenization treatment and filtration, and thus, filtered liquid containing 10-hydroxycamptothecin is obtained. The present invention has the characteristics of low synthesis cost and high yield of 10-hydroxycamptothecin.
Description
Affiliated technical field
The present invention relates to a kind of method of utilizing environmental factor to influence secondary metabolite, a kind of method that transforms 10-hydroxycamptothecine in the ultrasonic promotion bud of camptotheca seed of utilizing particularly is provided.
Background technology
At present, because natural 10-hydroxycamptothecine content in camptotheca seed is very low, 10-hydroxycamptothecine mainly is the approach acquisition by synthetic.China Intellectual Property Office disclosed " producing the method (CN 85 1 00520 A) of 10-hydroxycamptothecine with biotransformation method " on August 13rd, 1986, utilized aseptic bambermycin that camptothecine is converted into 10-hydroxycamptothecine, and transformation efficiency is 50%.The shortcoming of synthetic 10-hydroxycamptothecine is that toxic side effect is big, and utilizes aseptic bambermycin that camptothecine is converted into the complex process of 10-hydroxycamptothecine, cost height.
Summary of the invention
The objective of the invention is to overcome above-mentioned weak point, utilize the conversion of 10-hydroxycamptothecine in the ultrasonic promotion camptotheca seed germination process, and then obtain the extracting solution of the 10-hydroxycamptothecine of high-content, high yield.
In order to achieve the above object, the technical solution used in the present invention was: with camptotheca seed water planting 192-720 hour at normal temperatures, make its germination, the camptotheca seed that germinates is placed Vltrasonic device, add water again, make its submergence kind bud, at frequency 20-40KHz, under the temperature 20-35 ℃ of condition after ultrasonic radiation 10-30 minute, camplotheca acuminata kind bud after the radiation placed add 70-95% alcoholic acid refiner in advance and carry out kind of the fragmentation of bud tissue, homogenized and filter, obtain containing the filtrate of 10-hydroxycamptothecine.
Technology of the present invention is simple to operation, cost is low, and the content of 10-hydroxycamptothecine is equivalent to the dry camptotheca seed of the charging capacity of 1.5-2 ‰ in the gained extracting solution, and the content of 10-hydroxycamptothecine has increased 20-25 doubly than exsiccant camptotheca seed.
Embodiment
Below embodiments of the invention are described in further detail, the present invention was with camptotheca seed water planting 192-720 hour at normal temperatures, make its germination, the camptotheca seed that germinates is placed Vltrasonic device, add water again, make its submergence kind bud, at frequency 20-40KHz, under the temperature 20-35 ℃ of condition after ultrasonic radiation 10-30 minute, camplotheca acuminata kind bud after the radiation placed add 70-95% alcoholic acid refiner in advance and carry out kind of the fragmentation of bud tissue, homogenized and filter, obtain containing the filtrate of 10-hydroxycamptothecine.
Embodiment 1
With 2 kilograms of camptotheca seeds water planting 192 hours at normal temperatures, make its germination, the camptotheca seed that germinates is placed Vltrasonic device, add 4 kilograms in water again, make its submergence kind bud, at frequency 20KHz, under 25 ℃ of conditions of temperature, after the ultrasonic radiation 20 minutes, the camplotheca acuminata kind bud after the radiation placed add 5 liter of 70% alcoholic acid refiner in advance and carry out kind of the fragmentation of bud tissue, homogenized and filter, obtain containing the filtrate of 10-hydroxycamptothecine.Filtrate is centrifugal, gets supernatant liquor, measures through high performance liquid phase, and the content of 10-hydroxycamptothecine is 52.79mg/l in the filtrate, and the content of 10-hydroxycamptothecine has increased by 20 times than exsiccant camptotheca seed.
Embodiment 2
With 10 kilograms of camptotheca seeds water planting 750 hours at normal temperatures, make its germination, the camptotheca seed that germinates is placed Vltrasonic device, add 25 kilograms in water again, make its submergence kind bud, at frequency 40KHz, ultrasonic radiation is after 30 minutes under 20 ℃ of conditions of temperature, camplotheca acuminata kind bud after the radiation placed add 25 liter of 85% alcoholic acid refiner in advance and carry out kind of the fragmentation of bud tissue, homogenized and filter, obtain containing the filtrate of 10-hydroxycamptothecine.Filtrate is centrifugal, gets supernatant liquor, measures through high performance liquid phase, and the content of 10-hydroxycamptothecine is 26.39mg/l in the filtrate, and the content of 10-hydroxycamptothecine has increased by 25 times than exsiccant camptotheca seed.
Embodiment 3
With 15 kilograms of camptotheca seeds water planting 720 hours at normal temperatures, make its germination, the camptotheca seed that germinates is placed Vltrasonic device, add 30 kilograms in water again, make its submergence kind bud, at frequency 40KHz, ultrasonic radiation is after 10 minutes under 20 ℃ of conditions of temperature, camplotheca acuminata kind bud after the radiation placed add 30 liter of 95% alcoholic acid refiner in advance and carry out kind of the fragmentation of bud tissue, homogenized and filter, obtain containing the filtrate of 10-hydroxycamptothecine.Filtrate is centrifugal, gets supernatant liquor, measures through high performance liquid phase, and the content of 10-hydroxycamptothecine is 29.04mg/l in the filtrate, and the content of 10-hydroxycamptothecine has increased by 22 times than exsiccant camptotheca seed.
Claims (1)
1, promotes the method that 10-hydroxycamptothecine generates in the bud of camptotheca seed, it is characterized in that: with camptotheca seed water planting 192-720 hour at normal temperatures, make its germination, the camptotheca seed that germinates is placed Vltrasonic device, add water again, make its submergence kind bud, at frequency 20-40KHz, under the temperature 20-35 ℃ of condition after ultrasonic radiation 10-30 minute, camplotheca acuminata kind bud after the radiation placed add 70-95% alcoholic acid refiner in advance and carry out kind of the fragmentation of bud tissue, homogenized and filter, obtain containing 10-hydroxycamptothecine filtrate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB021329222A CN1183138C (en) | 2002-09-11 | 2002-09-11 | Process for promoting generation of 10-hydroxycamptothecin in the bud of camptotheca seed |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB021329222A CN1183138C (en) | 2002-09-11 | 2002-09-11 | Process for promoting generation of 10-hydroxycamptothecin in the bud of camptotheca seed |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1482129A CN1482129A (en) | 2004-03-17 |
| CN1183138C true CN1183138C (en) | 2005-01-05 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB021329222A Expired - Fee Related CN1183138C (en) | 2002-09-11 | 2002-09-11 | Process for promoting generation of 10-hydroxycamptothecin in the bud of camptotheca seed |
Country Status (1)
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| CN (1) | CN1183138C (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103875327B (en) * | 2014-03-25 | 2015-03-25 | 东北林业大学 | Camptotheca acuminata seed germination method |
| CN107043795B (en) * | 2017-02-04 | 2020-10-09 | 中国科学院成都生物研究所 | Method for producing camptothecin and 10-hydroxycamptothecin by using camptotheca acuminata suspension cells |
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2002
- 2002-09-11 CN CNB021329222A patent/CN1183138C/en not_active Expired - Fee Related
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| CN1482129A (en) | 2004-03-17 |
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