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Method for enhancing fermentation yield of ansamycin

A technology of increasing production and ansamycin, applied in the field of bioengineering, can solve the problems of complex medium components, difficult research mechanism, uncertain components, etc., and achieve the effect of convenient large-scale production, high fermentation level and simple operation

Inactive Publication Date: 2010-12-22
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the cost of this technique is high, the components of the medium are complicated, the components are uncertain, and the repeatability is poor, which brings difficulties to the research mechanism and great difficulties to the subsequent separation and extraction.

Method used

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  • Method for enhancing fermentation yield of ansamycin
  • Method for enhancing fermentation yield of ansamycin
  • Method for enhancing fermentation yield of ansamycin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The strain used: Actinosynnema pretiosum ATCC31565, which was purchased from the American Standard Biological Collection (ATCC) in Rockefeller, Maryland, USA.

[0028] Seed culture: The ingredients of the seed medium are 0.4% of yeast extract, 1% of maltose, and 0.4% of glucose. Glycerol tubes are inoculated in primary seed medium, cultured at 25-32°C for 24-60h, inoculated with 1.0-3.0% inoculation amount in secondary seed medium, and cultured at 25-32°C for 24-60h.

[0029] Fermentation culture: fermentation medium is yeast extract 1%, corn flour filtrate 2%, glucose 0.5%, glycerin 4%, dipotassium hydrogen phosphate 0.05%, heptahydrate and ferrous sulfate 0.0002%, calcium carbonate 0.5%, and then configure 1M magnesium sulfate aqueous solution is added to the above fermentation broth for fermentation, and the concentration of magnesium ions put into the shake flask is 0.1-20mM. 1.0-3.0% inoculation amount is inoculated in a fermentation shaker flask, fermented at 150...

Embodiment 2

[0033] The bacterial classification that adopts: with embodiment 1;

[0034] Seed culture: with embodiment 1;

[0035] Fermentation culture: Same as in Example 1, the corn flour filtrate was changed to buckwheat flour filtrate, a certain amount of magnesium ion solution was added to the initial material for fermentation, and the magnesium ion concentration after being put into the shake flask was 0.1-20mM. ;

[0036] Sample processing and detection method: the same as in Example 1.

[0037] Such as figure 2 As shown, the results show that when different concentrations of magnesium ions are added, the ansamycin production is greatly improved compared with the control. When the concentration of magnesium ions is 2mM, the effect is the best, up to 85mg / L, compared with the control Ratio increased by nearly 3 times.

Embodiment 3

[0039] The bacterial classification that adopts: with embodiment 1;

[0040] Seed culture: with embodiment 1;

[0041] Fermentation culture: same as in Example 1, adding 2mM magnesium ions at 0, 24, 48, and 72 hours respectively.

[0042] Sample processing and detection method: the same as in Example 1.

[0043] Such as image 3 As shown, the results show that the effect of adding magnesium ions is the most significant at 0h.

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Abstract

The invention discloses a method for enhancing fermentation yield of ansamycin, belonging to the technical field of bioengineering. The method comprises the following steps of inoculating glycerol to a seed culture medium in a melting way, carrying out culture operation, obtaining a two-stage seed after secondary culture, treating the two-stage seed by shake culture, adding a magnesium sulfate solution with the concentration of 1M into a fermentation shake flask for carrying out fermentation treatment, treating the fermentation shake flask by swinging culture, and realizing fermentation production. The invention has the advantages of high fermentation level, simple operation and low cost, and is beneficial to mass production.

Description

technical field [0001] The invention relates to a method in the technical field of bioengineering, in particular to a method for increasing the fermentation yield of ansamycin. Background technique [0002] Ansamitocin is a maytansine derivative produced by Actinosynnema pretiosum. Its core is a 19-C macrocyclic lactam ring, and C-3 is connected to carbon chains of different lengths. , the main active ingredient ansamycin AP-3 (AP-3), whose side chain is isobutyryl, 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, it has important application potential. [0003] The biosynthetic pathway of ansamycin includes a total of 48 genes (scattered in two gene clusters), of which asm2, 8, 18, 29, 31, 34, 39 and asm40 ...

Claims

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Application Information

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IPC IPC(8): C12P17/18C12R1/01
Inventor 钟建江贾永亮
Owner SHANGHAI JIAO TONG UNIV
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