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AspergillusflavusBP6T2 producing paclitaxel and application of aspergillusflavus

A technology of BP6T2 and paclitaxel, applied in the field of microorganisms, can solve the problems of expensive process, low yield, unstable yield, etc., and achieve the effect of expanding the scope of resources

Active Publication Date: 2017-07-21
曹军卫 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] (1) The method of chemical total synthesis of paclitaxel has been successful, but the whole process is very expensive and the yield is not high, so it is not an ideal way
[0008] (2) Although semi-synthetic is effective, it is difficult to separate and purify semi-synthetic precursors from plant tissues
[0009] (3) The steps required for plant tissue culture are relatively simple, but the yield of paclitaxel is low and the yield is unstable
Studies have shown that taxane compounds are also found in Taxusaceae Torreya plants, but the content is extremely low, and the paclitaxel content is lower than 0.003%. It is considered that the value of medicinal development is not large (Yi Guanmei, Qiu Yingchun. Research status and development [J]. Resources and Environment, 2013, 29(8): 844-848)

Method used

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  • AspergillusflavusBP6T2 producing paclitaxel and application of aspergillusflavus
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  • AspergillusflavusBP6T2 producing paclitaxel and application of aspergillusflavus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1: Separation and purification of the endophytic bacteria of Torreya bashanense

[0043] 1. Pretreatment of Torreya birch tree material

[0044] The roots, stems, leaves, and bark of Torreya bashanense were collected in Shennongjia National Geological Forest Park, Hubei Province, rinsed with sterile water, and blotted dry with absorbent paper; sterilized by 75% (v / v) alcohol for 5 minutes and 2% (v / v) hypochlorous acid disinfection for 8 minutes, then rinse with sterile water and cut the collected roots, stems and bark into small sections of about 2-3 cm with a sterile knife, and cut the leaves from the middle.

[0045] 2. Isolation of endophytic bacteria from Torreya bashanense

[0046] Take the pretreated materials from each part and place them on a PDA solid medium plate, and cultivate them at 28°C; PDA medium: 200g of potatoes, 20g of glucose, KH 2 PO 4 1.0g, MgSO 4 ·7H 2 O 0.5g, water 1000mL, add agar (15g / L) as PDA solid medium.

[0047] 3. Purif...

Embodiment 2

[0049] Embodiment 2: detection of endophytic fungus antibacterial activity

[0050] 1. After activating the test bacteria (Staphylococcus aureus, Escherichia coli) with LB liquid medium, spread them on the surface of LB solid medium respectively;

[0051] 2. Cut off the purified plant endophyte agar medium block obtained in Example 1, place and inoculate it in Staphylococcus aureus and Escherichia coli plates respectively, cultivate at 37° C. for 1-2 days, and observe the bacteriostatic zone around the agar block.

[0052] By the above-mentioned method, screening obtains a strain that simultaneously has anti-Staphylococcus aureus (G + ) and Escherichia coli (G - ) active strain, denoted as BP6T2.

Embodiment 3

[0053] Example 3: Identification of endophytic fungi BP6T2

[0054] Through the observation of plate culture characteristics, the colony color of the strain is grass green, and the colony texture is velvet ( figure 2 ); under the microscope, the conidia heads are loose and radial, the top capsule is nearly spherical, and the small stalks exist in a single layer and double layer ( image 3 ), according to "Chinese Mycology Fifth Aspergillus and Its Related Sexual Types" (Edited by Qi Zutong, Chinese Mycology (Volume V) Aspergillus and Its Related Sexual Types [M]. Beijing: Science Press, 1997 ,5~10.) can be preliminarily identified as Aspergillus flavus (Aspergillus flavus) BP6T2.

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Abstract

The invention discloses an endophytewhich produces paclitaxel, is separated and screened out fromtissue of taxaceaetorreyafargesii, is named as aspergillusflavusBP6T2through classification and determination, and has a preservation number of CCTCCNO:M2016318. The strain has an activity of resisting staphylococcus aureus and escherichia coli, and an extract from fermentation liquor can reach 93mg / L, wherein the relative percentage content of paclitaxel is 8.4%, and the yield is 7.81mg / L and is higher than the yield disclosed in the prior art; and the extract of the fermentation liquor of the aspergillusflavusBP6T2 is proved by a CCK-8 reagent method to have an effect of inhibiting Hela tumor cells, and the inhibition rate can reach 66.20%.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to an Aspergillus flavus BP6T2 isolated from Torreya barbata. The bacterial strain has the characteristic of high-yielding paclitaxel, and also relates to the application of Aspergillus flavus BP6T2 in treating tumor cells. Background technique [0002] Paclitaxel (Paclitaxel) alias taxol, porphyrin, special element, molecular weight 853.92, molecular formula C 47 h 51 NO 14 ( figure 1 ), is a cell mitosis inhibitor, which is used in cancer therapy (Wani MC, Taylor HL, Wall ME, et al. Plantantitumor agents. VI. The isolation and structure of taxol, a novel antileukemic and antitumor agent from Taxus brevifolia[J]. J Am Chem Soc, 1971, 93:2325-2327.). [0003] Paclitaxel and its analogs are considered to be highly effective, broad-spectrum, low-toxic anticancer drugs, especially the best drugs for the treatment of ovarian cancer, uterine cancer and breast cancer. , ...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P17/02C12R1/67
CPCC12P17/02C12N1/145C12R2001/67
Inventor 曹军卫涂毅金卫华
Owner 曹军卫
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