A taxol-producing endophytic fungus Aspergillus aculeatus tax-6 and its application

Abstract

The invention discloses a high-efficiency paclitaxel-producing fungus Aspergillus aculeatus Tax-6 and its application. The high-efficiency paclitaxel-producing fungus is named Aspergillus aculeatus ( Aspergillus aculeatinus ) Tax‑6, deposited in the China Center for Type Culture Collection, the deposit number is CCTCC M 2016614, the deposit date is November 3, 2016, and the deposit address is Wuhan University, Wuhan, China. The present invention cultivates Tax‑6 through a liquid fermentation process and then extracts paclitaxel in the fermentation broth, the highest content reaches 334.92 µg / L and the culture time only needs 7 days. The present invention utilizes the endophytic fungus of yew bark to ferment paclitaxel to produce paclitaxel, which not only produces paclitaxel, a high-efficiency anticancer drug, but also protects the rare species of yew.

Description

technical field

[0001] The invention relates to the technical field of drug production by plant endophytic fungi, in particular to a high-efficiency paclitaxel-producing fungus Aspergillus aculeatus Tax-6 and its application. Background technique

[0002] Cancer is one of the three major killers threatening human health in the 21st century. According to the survey by the National Cancer Registry Center, since 2013, there have been more than 3 million new cancer cases in my country every year. Therefore, the task of finding highly effective anticancer drugs is also imminent. Paclitaxel ( Taxol ) originally from the genus Taxus ( Taxus spp. ) is a diterpenoid isolated from plants with a unique anticancer effect. It acts on cell microtubules and induces and stabilizes microtubules by binding to amino acids 31 and 217-231 at the N-terminal of tubulin. Tubulin polymerization, inhibit its depolymerization, increase the polymerization speed, so that the vascular bundle can not b...

Examples

Embodiment 1

[0025] Liquid fermentation culture: take out from the refrigerator at 4°C and store Aspergillus aculeatinus The PDA plate of Tax-6 was placed in a biochemical incubator at 28°C, and the strains were activated for 2-3 days. On the sterilizer, add about 20 mL of sterile water to the activated plate, gently scrape the spores on the mycelium into the sterile water with an inoculation needle, mix well, and filter aseptically to make a spore suspension. liquid (about 1 x 10 7 spores / mL). Draw 2 mL of spore suspension with a sterile pipette tip and inoculate it into a 250 mL Erlenmeyer flask containing 100 mL of PDA solid medium, and place the inoculated Erlenmeyer flask on a shaker at 120 rpm at 30°C for cultivation.

[0026] Extraction of paclitaxel: filter the fermentation broth with four layers of gauze to take the liquid part, add 10 mL of n-hexane to extract for 20 min to remove impurities such as fatty acids, and keep the water phase. Paclitaxel in the fermentation broth ...

technical field

[0001] The invention relates to the technical field of drug production by plant endophytic fungi, in particular to a high-efficiency paclitaxel-producing fungus Aspergillus aculeatus Tax-6 and its application. Background technique

[0002] Cancer is one of the three major killers threatening human health in the 21st century. According to the survey by the National Cancer Registry Center, since 2013, there have been more than 3 million new cancer cases in my country every year. Therefore, the task of finding highly effective anticancer drugs is also imminent. Paclitaxel ( Taxol ) originally from the genus Taxus ( Taxus spp. ) is a diterpenoid isolated from plants with a unique anticancer effect. It acts on cell microtubules and induces and stabilizes microtubules by binding to amino acids 31 and 217-231 at the N-terminal of tubulin. Tubulin polymerization, inhibit its depolymerization, increase the polymerization speed, so that the vascular bundle can not b...

Examples

Embodiment 1

[0025] Liquid fermentation culture: take out from the refrigerator at 4°C and store Aspergillus aculeatinus The PDA plate of Tax-6 was placed in a biochemical incubator at 28°C, and the strains were activated for 2-3 days. On the sterilizer, add about 20 mL of sterile water to the activated plate, gently scrape the spores on the mycelium into the sterile water with an inoculation needle, mix well, and filter aseptically to make a spore suspension. liquid (about 1 x 10 7 spores / mL). Draw 2 mL of spore suspension with a sterile pipette tip and inoculate it into a 250 mL Erlenmeyer flask containing 100 mL of PDA solid medium, and place the inoculated Erlenmeyer flask on a shaker at 120 rpm at 30°C for cultivation.

[0026] Extraction of paclitaxel: filter the fermentation broth with four layers of gauze to take the liquid part, add 10 mL of n-hexane to extract for 20 min to remove impurities such as fatty acids, and keep the water phase. Paclitaxel in the fermentation broth ...

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