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Solid fermentation method of ansamitocin

A technology of solid fermentation and anisomycin, applied in microorganism-based methods, fermentation, biochemical equipment and methods, etc., can solve problems such as difficulties in industrial fermentation, and achieve the effects of simple sample processing and low energy consumption

Inactive Publication Date: 2008-12-10
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, the U.S. Patent No. US6573074 describes a method for purifying ansamectin. The method is to use toluene or xylene to crudely extract the product from the fermentation broth and concentrate it. After petroleum ether precipitation, it passes through various columns. Chromatography further separates, purifies and crystallizes, and obtains ansamitocin with a certain yield and purity after secondary crystallization with ethyl acetate and heptane as solvents. However, this method involves a variety of organic solvents and extraction methods, which is difficult for large-scale industrial Fermentation poses certain difficulties

Method used

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  • Solid fermentation method of ansamitocin
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  • Solid fermentation method of ansamitocin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Comparison of the ansamitocin production of YMG medium solid fermentation and YMG medium liquid fermentation:

[0021] Seed and fermentation culture: the seed medium is YMG medium, and the weight percentage of its components is: 0.4% of yeast extract, 1% of maltose, 0.4% of glucose, 1.5% of agar powder and 96.7% of distilled water, and the diameter of the petri dish is 10cm. The filling volume is 15ml. Use the high-concentration thalline streak YMG medium plate preserved in 10.3% sucrose, activate it upside down at 28°C for 9 days, and inoculate it into a 250ml shake flask containing 30ml of YMG medium with an inoculation spatula, at 28°C, 220 RPM for 2 days, press 10 2 The inoculum amount of CFU / ml was spread on the YMG medium plate covered with the regenerated cellulose film support, and cultured at 28°C for 7 days. At the same time, the same inoculum amount was inoculated into a 250ml shaker containing 30ml of YMG medium with an inoculation spatula. In the bottle, ...

Embodiment 2

[0025] Comparison of solid-liquid fermentation production of ansamitocin with different media compositions using a renewable cellulose film support:

[0026] Seed culture: the seed medium is YMG medium, and the mass percentage of its components is: 0.4% of yeast extract, 1% of maltose, 0.4% of glucose, 1.5% of agar powder and 96.7% of distilled water. The diameter of the petri dish is 10cm. Liquid volume 15ml. 10.3% sucrose-preserved high-concentration thalline streaked YMG plate was activated upside down at 28°C for 5 days, inoculated into a 250ml shaker flask containing 30ml of YMG medium with an inoculation spatula, and cultured at 220 rpm at 28°C for 2 day, after serial dilution, with 10 6 The concentration of CFU / ml was spread on the YMG plate covered with regenerated cellulose film support, and cultured for 9 days at 28°C. At the same time, the same inoculum was inoculated into a 250ml shake flask containing 30ml of YMG medium, at 28°C, 28 Cultivate at 220 rpm for 9 da...

Embodiment 3

[0036] Comparison of the effect of different inoculum densities on ansamitocin in solid state fermentation using a renewable cellulose film support.

[0037] Seed and fermentation culture: The mass percentage of each component of seed culture is: yeast extract 0.4%, maltose 1%, glucose 0.4%, agar powder 1.5% and distilled water 96.7%, the diameter of the petri dish is 10cm, and the liquid volume is 15ml. 10.3% sucrose-preserved high-concentration thalline streaked YMG plates were inverted at 28°C for 2 days, and a small piece was inoculated into a 250ml shaker flask filled with 30ml of YMG medium with an inoculation shovel, at 28°C at 220 The rotation speed of rev / min was cultivated for 2 days, and after serial dilution, the 4 The concentration of CFU / ml was spread on the YMG plate covered with the regenerated cellulose film support, and cultured for 10 days at 28°C.

[0038] The method of attaching the regenerated cellulose film support is as described in Example 1.

[0039...

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Abstract

The invention discloses a solid fermentation method for ansamitocin. The method comprises the following steps of: step one, seed culture: a thallophytic streak culture medium plate which is freezingly preserved in a saccharose solution and produces the ansamitocin is subject to inversion activation for 2 to 9 days at a temperature of 28 DEG C and then is transferred to a seed culture medium, and a seed liquid is cultured at a temperature of 28 DEG C; and step two, solid fermentation: the seed liquid obtained in the step one is diluted to a concentration between 10<2> and 10<6>CFU / ml and is coated to a fermentation culture medium plate paved with a regenerative cellulose membrane support to be cultured for 7 to 10 days at a temperature of 28 DEG C. Compared with the prior liquid fermentation method, the method has the advantages that: the output of the ansamitocin is more than 2 times higher, and the unit cell content is near 4 times higher; at the same time, the solid fermentation has small energy consumption, the treatment of a sample is simple, thereby contributing to the industrialized preparation of the ansamitocin.

Description

technical field [0001] The invention relates to a fermentation method in the field of bioengineering and technology, in particular to a solid fermentation method of ansamectin. Background technique [0002] Ansamitocin (Ansamitocin) is a maytansine derivative derived from Actinosynnema pretiosum. Its core is a 19-C macrocyclic lactam ring, and C-3 is connected to carbon chains of different lengths. , the side chain of the main active ingredient AP-3 is isobutyryl, which can inhibit non-leukemic leukemia cell lines and human solid tumors at low concentrations. Due to gastrointestinal side effects and neurotoxicity, its research is still in the second clinical stage. However, since ansamitocin can be used as an antibody-toxin conjugate and immune conjugate in the first clinical phase, research in various aspects is still in progress. [0003] Solid fermentation is a kind of fermentation close to the natural state. It has many differences from liquid submerged fermentation, a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/18C12R1/01
Inventor 钟建江林锦霞
Owner SHANGHAI JIAO TONG UNIV
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