A method based on porous ceramic adsorption and immobilization of S. cellulosus fermentation

A technology of cellulosystis and porous ceramics, applied in the direction of microorganism-based methods, immobilized on or in inorganic carriers, fermentation, etc., can solve the problems of cell autolysis, unstable metabolic state, inconsistent state of internal and external cells, etc. , to achieve the effects of alleviating feedback inhibition, reducing production costs, and less wear and tear

Inactive Publication Date: 2014-10-01
SHAANXI UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

They tend to grow, metabolize and produce epothilone well on solid media, but their metabolic behavior often changes when growing in liquid media; for example, cells grow in clumps, and the states of inner and outer cells are inconsistent; cells exist Autolysis phenomenon; unstable metabolic state, etc.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] A method for fermenting and producing epothilone B based on porous ceramic adsorption and immobilization of S. cellulosus, comprising the following steps:

[0057] 1) Diatomite-based porous ceramics with a size of Ф20×10mm were sterilized at 150°C for 30 minutes at high temperature. After natural cooling, they were put into a conical flask and fermented liquid was added. After the liquid surface completely covered the porous ceramic samples, it was about 50mm, shake slowly at room temperature for 1 hour, let it stand for 2 hours, pour off the fermentation broth, and dry the porous ceramics at 105°C for later use.

[0058] 2) Inoculate a ring of fresh Sorangium cellulosum on M26 solid medium, and culture in a constant temperature incubator at 30°C for 72 hours;

[0059] Inoculate the cultured bacteria into 100ml of M26 liquid medium, culture at 30°C, 200rpm shaker for 72h;

[0060] After the cultivation, part of the liquid was discarded, and the remaining liquid was tra...

Embodiment 2

[0064] A method for fermenting and producing epothilone B based on porous ceramic adsorption and immobilization of S. cellulosus, comprising the following steps:

[0065] 1) Diatomite-based porous ceramics with a size of Ф20×10mm were sterilized at 150°C for 30 minutes at high temperature. After natural cooling, they were put into a conical flask and fermented liquid was added. After the liquid surface completely covered the porous ceramic samples, it was about 50mm, shake slowly at room temperature for 1 hour, let it stand for 2 hours, pour off the fermentation broth, and dry the porous ceramics at 105°C for later use.

[0066] 2) Prepare the bacterial suspension at a certain concentration with the method in Example 1.

[0067] 3) Take 5ml of the diluted bacterial suspension, put it into a 500mL culture bottle containing 100mL of fermentation broth and 1g of porous ceramics, and incubate on a shaker at 28°C and 180rpm for 96h.

Embodiment 3

[0069] A method for fermenting and producing epothilone B based on porous ceramic adsorption and immobilization of S. cellulosus, comprising the following steps:

[0070] 1) Diatomite-based porous ceramics with a size of Ф20×10mm were sterilized at 150°C for 30 minutes at high temperature. After natural cooling, they were put into a conical flask and fermented liquid was added. After the liquid surface completely covered the porous ceramic samples, it was about 50mm, shake slowly at room temperature for 1 hour, let it stand for 2 hours, pour off the fermentation broth, and dry the porous ceramics at 105°C for later use.

[0071] 2) Prepare a certain concentration of bacterial suspension with the method in Example 1.

[0072] 3) Take 10ml of the diluted bacterial suspension, put it into a 500mL culture bottle containing 100mL of fermentation broth and 1.5g of porous ceramics, and incubate on a shaking table at 30°C and 220rpm for 96h.

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Abstract

The invention discloses a method for absorbing and fixing sorangium cellulosum for fermentation based on porous ceramics and belongs to the field of microbial fermentation pharmacy. The method comprises the steps that the prepared porous ceramics are processed by fermentation liquid to be dried for stand-by application; after sorangium cellulosum is cultivated through an M26 solid and a liquid medium respectively, bacterium suspension is formed; and after dilution, the bacterium suspension is inoculated in the fermentation liquid, the processed porous ceramics are added into the fermentation liquid, and shake cultivation is performed. The method for absorbing and fixing an epothilone production bacterial strain based on kieselguhr-based porous ceramics is provided by the invention, when epothilone B is in the fermentation production, sorangium cellulosum, which is the production bacterial strain of the epothilone B, is fixed, so that a solid attachment surface can be provided for a slime bacteria cell growing in a liquid environment, the method has the advantages that the fermentation production yield can be increased, the production cost of epothilone which is an anti-tumor medicine is lowered, and the operation steps are simple and easy to implement, therefore, the method can be amplified to the industrial production and establishes an experimental basis for the industrial production of epothilone.

Description

technical field [0001] The invention belongs to the technical field of Sotropha cellulosus fermentation technology, and relates to a fermentation method for absorbing and immobilizing Sotropha cellulosus based on porous ceramics. Background technique [0002] Epothilone B (Epothilone B, EPO-906) is a drug produced by S. cellulosus, which has entered clinical anti-tumor treatment research. At present, the fermentation yield of epothilone is very low, mainly due to two reasons. The first is that epothilones have a toxic inhibitory effect on their production strains, which limits the fermentation yield of epothilones to a large extent. Although the macroporous adsorption resin XAD-16 can be used to solve this difficulty, the non-specific adsorption of the resin to other substances in the fermentation broth also brings new difficulties to the improvement of the yield of epothilone. Second, the complex cellular behavior of S. cellulosus. They tend to grow, metabolize and produ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P17/18C12N11/14C12R1/01
Inventor 龚国利刘丽丽王娜
Owner SHAANXI UNIV OF SCI & TECH
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