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Fermentation technique of A40926

A fermentation process and fermentation tank technology, applied in the biological field, can solve the problems of low fermentation yield and failure to meet production requirements, and achieve the effects of high fermentation level, low cost and simple operation

Inactive Publication Date: 2015-04-08
SHANGHAI INST OF PHARMA IND CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to overcome the low fermentation yield of the A40926 fermentation process in the prior art, which is limited to simple shake flask experiments and cannot meet the production requirements, and provides a simple operation, low cost, and Large-scale production, the fermentation process of the key intermediate A40926 that can produce up to 720mg / L dalbavancin

Method used

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  • Fermentation technique of A40926

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Inoculate the Nonomuraea genus Actinomycetes ATCC 39727 (Nonomuraea.sp ATCC 39727) stored on the slant plate into the primary seed medium of the shake flask: beef extract 0.3%, casein 0.5%, oat flour 3%, calcium carbonate 0.4%, pH 7.0, cultured at 30°C for 72h, shaker speed 240rpm; 5% inoculum amount was inserted into the secondary seed medium (the same level seed medium) of the shake flask, and cultured on a rotary shaker at 28°C for 72h , Shaker speed 240rpm.

[0023] Afterwards, according to the 8% inoculum amount, inoculate in a 50L fermenter for fermentation, the actual liquid volume is 28L, and cultivate at 26°C for 226h. Wherein, the stirring speed is 200 rpm, the tank pressure is 0.02 MPa, and the ventilation ratio is 1:1 V / V·min.

[0024] Fermentation medium is prepared according to conventional methods, and every liter comprises glucose 4.5%, ammonium sulfate 0.83%, potassium dihydrogen phosphate 0.15%, magnesium sulfate heptahydrate 0.1%, yeast extract 0.1%,...

Embodiment 2

[0028] Inoculate the Nonomuraea genus Actinomycetes ATCC 39727 (Nonomuraea.sp ATCC 39727) preserved on the slant plate into the primary seed medium of the shake flask: beef extract 0.4%, casein 0.2%, oat flour 2%, calcium carbonate 0.2%, pH 7.0, cultured at 30°C for 75h, shaker speed 240rpm; 5% inoculum size was inserted into the shake flask secondary seed medium (seed medium of the same level), and cultivated at 28°C for 72h on a rotary shaker , Shaker speed 240rpm.

[0029] Afterwards, according to the 6% inoculum amount, inoculate in a 50L fermenter for fermentation, the actual liquid volume is 28L, and cultivate at 28°C for 215h. Wherein, the stirring speed is 240rpm, the tank pressure is 0.03MPa, and the ventilation ratio is 1:1.2V / V·min.

[0030] The fermentation medium is prepared according to conventional methods, and each liter contains 3% corn starch, 0.5% glucose, 1% sucrose, 3% soybean cake powder, 1.5% fish meal peptone, 0.4% casein, 0.1% L-valine, carbonic acid ...

Embodiment 3

[0033] Inoculate the Nonomuraea genus Actinomycetes ATCC 39727 (Nonomuraea.sp ATCC 39727) preserved on the slant plate into the primary seed medium of the shake flask: 0.5% beef extract, 0.5% casein, 4% oat flour, calcium carbonate 0.5%, pH 7.0, cultured at 30°C for 75h, shaker speed 240rpm; 5% inoculum was inserted into the shake flask secondary seed medium (seed medium of the same level), and cultivated at 28°C for 72h on a rotary shaker , Shaker speed 240rpm.

[0034] According to the 8% inoculum amount, it is inoculated in a 50L fermenter for fermentation, and the actual liquid volume is 28L. Incubate at 27°C for 218h. The stirring speed is 250rpm, the tank pressure is 0.05MPaMPa, and the ventilation ratio is 1:1V / V·min

[0035] The fermentation medium is prepared according to conventional methods, and each liter contains 5% corn starch, 0.75% glucose, 1.5% sucrose, 3.5% soybean cake powder, 1.25% fish meal peptone, 0.45% casein, 0.11% L-valine, carbonic acid Calcium 0....

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Abstract

The invention discloses a fermentation technique of A40926, which comprises the following steps: inoculating a strain capable of producing A40926 in a fermentation culture medium, and carrying out fermentation and culture, wherein after the fermentation and culture are carried out for 72-112 hours, a solution containing 0.5-1.2 wt% of glucose (on the basis of the fermentation culture medium), 0.3-0.8 wt% of peptone and 0.05-0.12 wt% of L-valine is fed. The technique is simple to operate and low in cost, can implement large-scale production, and can maximally produce 720 mg / L Dalbavancin intermediate A40926.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a fermentation process of A40926. Background technique [0002] Dalbavancin (Dalbavancin) intermediate A40926 is a glycopeptide substance discovered by scientists in 1984 when they cultivated and isolated Actinomadura from soil. In 2003, the producing bacteria of A40926 were classified as Nonomuraea genus Actinomycetes ATCC 39727 (Nonomuraea.sp ATCC 39727). A40926 includes 5 main components: PA, PB, A, B0 and B1, where B0 and B1 are collectively referred to as component B. A new type of glycopeptide antibacterial drug dalbavancin can be obtained after chemical structure modification of A40926B component. [0003] Dalbavancin was developed by Vicuron and is currently in phase III clinical trials. Studies have found that the mechanism of action of dalbavancin is the same as that of vancomycin and teicoplanin. It inhibits the biosynthesis of the cell wall of Gram+ bacteri...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/00C12R1/01
Inventor 胡海峰陈昌发张长清朱宝泉
Owner SHANGHAI INST OF PHARMA IND CO LTD
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