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Method for increasing fermentation yield of epothilone by using competitive microorganism and application thereof

A microbial fermentation and epothilone technology, applied in the field of microorganisms, can solve the problems of increasing production, low potential, and low efficiency of strain improvement methods

Inactive Publication Date: 2012-04-04
QILU UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there are mainly two strategies for increasing the fermentation yield of microbial drugs, namely, traditional breeding and optimization of fermentation conditions. The further use of traditional breeding methods for industrial strains has little potential to greatly increase the yield, while the genetic background of other strains is not clear or the operating methods are not clear. For perfect strains such as S. cellulosus, the efficiency of strain improvement is not high; optimization of fermentation conditions, as another key technology to increase yield, has also been introduced in almost all fermentation production processes, but the strains have limited ability to metabolize nutrients , it is impossible for us to increase the yield without limit by optimizing the fermentation conditions
When some strains undergo continuous breeding or optimization of fermentation conditions, the yield reaches a certain bottleneck, and it is difficult to make a breakthrough by using these two methods

Method used

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  • Method for increasing fermentation yield of epothilone by using competitive microorganism and application thereof
  • Method for increasing fermentation yield of epothilone by using competitive microorganism and application thereof
  • Method for increasing fermentation yield of epothilone by using competitive microorganism and application thereof

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Experimental program
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Effect test

Embodiment 1

[0036]1. The preparation method of Rhizopus aurizopus ATCC56015 fermentation product, the steps are as follows:

[0037] (1) Take the preserved strain and inoculate it into the activation medium, culture conditions: temperature 29°C, culture for 3 days for activation.

[0038] (2) Take the activated Rhizopus strain and inoculate it in the nutrient medium, and culture it for 4 days under the culture conditions of 30° C. and 150 r / min.

[0039] (3) Add ethyl acetate to the fermentation broth at a volume ratio of 1:1, stir and extract at 200 r / min for 3 hours, repeat 2 times, combine the ethyl acetate phases, and vacuum dry.

[0040] (4) Dissolving the dry product obtained in step (3) with acetone at 1 / 200 of the fermentation broth volume, and the supernatant after microfiltration is the metabolite of the competing strain.

[0041] The activation medium in the step 1 is: potato (peeled): 200g, glucose: 20g, agar: 20g, water: 1000mL, and the pH value is natural.

[0042] The nut...

Embodiment 2

[0050] 1. The method for preparing the metabolite of Rhizopus aurizopus ATCC56015, the steps are as follows:

[0051] (1) Inoculate the rhizopus strain into the activation medium, culture conditions: temperature 30°C, culture for 3 days for activation.

[0052] (2) Take the activated Rhizopus strain and inoculate it in the seed medium, and cultivate it for 4 days under the culture conditions of 30° C. and 150 r / min.

[0053] (3) Add ethyl acetate to the fermentation broth at a volume ratio of 1:1, stir and extract at 200 r / min for 3 hours, repeat 2 times, combine the ethyl acetate phases, and vacuum dry.

[0054] (4) Dissolve the dry product obtained in step (3) with 1 / 200 of the fermentation broth volume in methanol, centrifuge at 10,000 r / min for 10 min and the supernatant is the metabolite of the competing strain.

[0055] The activation medium in the step 1 is: potato (peeled): 200g, glucose: 20g, agar: 20g, water: 1000mL, and the pH value is natural.

[0056] The nutrie...

Embodiment 3

[0064] 1. The preparation method of the metabolite of Penicillium decumbens CGMCC 4075, the steps are as follows:

[0065] (1) Inoculate the Penicillium strain into the activation medium, culture conditions: temperature 30°C, culture for 3 days for activation.

[0066] (2) Take the activated Penicillium strain and inoculate it in the nutrient medium, and culture it for 4 days under the culture conditions of 30° C. and 150 r / min.

[0067] (3) Add ethyl acetate to the fermentation broth at a volume ratio of 1:1, stir and extract at 200 r / min for 3 hours, repeat 2 times, combine the ethyl acetate phases, and vacuum dry.

[0068] (4) Dissolve the dry product obtained in step (3) with 1 / 200 of the fermentation broth volume in ethanol, centrifuge at 10,000 r / min for 10 min, and the supernatant is the metabolite of the competing microorganisms.

[0069] The activation medium in the step 1 is: potato (peeled): 200g, glucose: 20g, agar: 20g, water: 1000mL, and the pH value is natural....

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Abstract

The invention discloses a method for increasing the fermentation yield of epothilone by using the metabolin of competitive microorganism namely rhizopus arrhizus or penicillium decumbens of sorangium cellulosum to induce. The method provided by the invention comprises the following steps of: cultivating the competitive microorganism, and extracting the metabolin of the competitive microorganism; adding the metabolin into a sorangium cellulosum fermentation medium, activating epothilone to produce related regulator genes inside the strain by using the stimulation of the external additive, and further stimulating the synthesis of epothilone. Detection results show that the fermentation yield of epothilone can be increased by 46.3-58.7% as compared with original conditions after the method is used, and the method has important theoretical significance and economic value.

Description

technical field [0001] The invention relates to a method for using competitive microorganisms and metabolites thereof to induce Sorangium cellulosum to increase the fermentation yield of epothilone and its application in antibiotic fermentation production, belonging to the technical field of microorganisms. Background technique [0002] Epothilones are a class of macrolide compounds synthesized by the myxobacterium S. cellulosus, which have the activity of promoting microtubule polymerization, and their mechanism of action is similar to paclitaxel Paclitaxel and its analogues are currently the most successful anti-tumor chemotherapy drugs in clinical practice. They are used in the treatment of solid cancers such as ovarian cancer, thymus cancer, lung cancer and liver cancer. Although paclitaxel is widely used, it still has some disadvantages, such as low water solubility, obvious toxic and side effects, easy to produce drug resistance, etc., which limit its application to a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P39/00C12P17/18C12R1/01C12R1/80C12R1/845
Inventor 赵林刘新利李亚伟
Owner QILU UNIV OF TECH
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