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Screening of new strain capable of producing camptothecin and method for preparing camptothecin

A technology of camptothecin and bacterial strains, which is applied in the field of preparing camptothecin, can solve the problems of long synthetic route, high cost, and low extraction yield, achieve the effects of mild reaction conditions, solve the problem of drug source, and simple operation

Inactive Publication Date: 2012-04-18
INST OF PROCESS ENG CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the main production method of camptothecin is to extract from camptothecin, and the content of camptothecin in camptothecin is very low, generally 0.01% to 0.1%, resulting in low extraction rate and high cost, and camptothecin is a national Level II focuses on the protection of wild plants, and a large number of felling of camphor trees will easily lead to vegetation damage, ecological imbalance and other problems
The chemical total synthesis of camptothecin has been successful, but it cannot be put into industrial production due to long synthetic route, low yield and high cost

Method used

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  • Screening of new strain capable of producing camptothecin and method for preparing camptothecin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Screen the new bacterial species producing camptothecin from camptothecin bark, the specific experimental steps are as follows:

[0023] 1. Rinse the surface dust of camphor tree bark with sterile water first, then soak it with ethanol with a volume ratio of 75% for 30s, wash off the surface ethanol with sterile water, and then soak it with mercuric chloride with a mass fraction of 0.1% for 30s. Rinse with sterile water for 3 to 5 times, and then cut it into small pieces of 5mm×5mm with a sterile knife;

[0024] 2. Inoculate the above materials on a PDA medium plate and incubate at a constant temperature of 28-30°C; when colonies grow obviously around the sample, pick colonies of different shapes and transfer them to the slope of the PDA medium, purify and cultivate at 28-30°C, and store spare;

[0025] 3. Strain activation: Inoculate the isolated endophytic fungi in the activation medium: glucose 20g / L, peptone 10g / L, magnesium sulfate 5g / L, potassium dihydrogen phosp...

Embodiment 2

[0029] Screen the new bacterial species producing camptothecin from the fruit of camptothecin, and the specific experimental steps are as follows:

[0030] 1. Rinse the surface dust of the camphor tree fruit with sterile water, then soak it in 75% ethanol by volume for 30s, wash off the surface ethanol with sterile water, then soak it in mercuric chloride with a mass fraction of 0.1% for 30s, sterile Rinse with water for 3 to 5 times, and then cut it into small pieces of 5mm×5mm with a sterile knife;

[0031] 2. Inoculate the above materials on a PDA medium plate and incubate at a constant temperature of 28-30°C; when colonies grow obviously around the sample, pick colonies of different shapes and transfer them to the slope of the PDA medium, purify and cultivate at 28-30°C, and store spare;

[0032] 3. Strain activation: Inoculate the isolated endophytic fungi in the activation medium: glucose 20g / L, peptone 10g / L, magnesium sulfate 5g / L, potassium dihydrogen phosphate 5g / L,...

Embodiment 3

[0036] Screen the new bacterial species producing camptothecin from camptothecin bark, the specific experimental steps are as follows:

[0037] 1. Rinse the surface dust of camphor tree bark with sterile water first, then soak it with ethanol with a volume ratio of 75% for 30s, wash off the surface ethanol with sterile water, and then soak it with mercuric chloride with a mass fraction of 0.1% for 30s. Rinse with sterile water for 3 to 5 times, and then cut it into small pieces of 5mm×5mm with a sterile knife;

[0038] 2. Inoculate the above materials on a PDA medium plate and incubate at a constant temperature of 28-30°C; when colonies grow obviously around the sample, pick colonies of different shapes and transfer them to the slope of the PDA medium, purify and cultivate at 28-30°C, and store spare;

[0039] 3. Strain activation: Inoculate the isolated endophytic fungi in the activation medium: glucose 20g / L, peptone 10g / L, magnesium sulfate 5g / L, potassium dihydrogen phosp...

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Abstract

The invention discloses a new strain capable of producing camptothecin and a method for preparing camptothecin by using the same. The new strain capable of producing the camptothecin-Phomopsis sp. B29 is conserved in China General Microbiological Culture Collection Center (CGMCC) on July 14, 2010, and has a conservation number of CGMCC No. 3988. The camptothecin is indole alkaloid with antitumor activity and is extracted from Camptotheca acuminate-nyssaceae plant of China, and the camptothecin has good curative effect on malignant tumors such as colon cancer, ovarian cancer and the like. Direct extraction of the camptothecin from the Camptotheca acuminate causes vegetation deterioration, ecological unbalance and other problems, and a chemical synthesis method has the disadvantages of long route, low yield and high cost; and aiming at the problems, endophytic fungi are separated and purified from the bark and fruit of the Camptotheca acuminate, and the strain capable of producing the camptothecin is screened from the endophytic fungi and then is used for preparing the camptothecin through fermentation. The new strain provided by the invention can be taken as a new medicine source for producing the camptothecin; and the method for preparing the camptothecin by fermenting the new strain is convenient to operate and has the advantage of low cost.

Description

technical field [0001] The invention belongs to the field of separation, screening and microbial fermentation of plant endophytes, in particular to a method for isolating and purifying endophytes from camptotheca japonica, fermenting and screening camptothecin-producing strains, and preparing camptothecin from the screened strains. Background technique [0002] Camptothecin is an indole alkaloid with anti-tumor activity extracted from Camptotheca acuminate by Wall et al. in 1966. At present, the main production method of camptothecin is to extract from camptothecin, and the content of camptothecin in camptothecin is very low, generally 0.01% to 0.1%, resulting in low extraction rate and high cost, and camptothecin is a national Level II focuses on the protection of wild plants, and cutting down a large number of camphor trees will easily lead to vegetation damage and ecological imbalance. The chemical total synthesis of camptothecin has been successful, but it cannot be put...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12P17/18C12R1/645
Inventor 陈洪章王玉美
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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