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Technology for realizing high-yield polyoxins by continuous fermentation coupled with separation

A multi-antibiotics and process technology, which is applied in the process of continuous fermentation and separation coupling to achieve high-yield multi-antibiotics, the field of fermentation and separation coupling control, can solve the problems of low titer and long cycle of multi-antibiotic fermentation, and achieves reduced energy consumption and shortened cycle. , the effect of improving production efficiency

Inactive Publication Date: 2016-08-03
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The technical problem to be solved by the present invention is to provide a process of continuous fermentation and separation coupling to realize high-yield multi-antibiotics, so as to solve the problems of low fermentation titer and long cycle of multi-antibiotics caused by product inhibition

Method used

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  • Technology for realizing high-yield polyoxins by continuous fermentation coupled with separation
  • Technology for realizing high-yield polyoxins by continuous fermentation coupled with separation

Examples

Experimental program
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Effect test

Embodiment 1

[0040] Seed medium: sucrose 12g / L, yeast extract 8g / L, KH 2 PO 4 0.2g / L, CaCO 3 0.1g / L. Pick a single colony of Streptomyces aureus chromogenicum NJYHYWG66302 from the plate medium and inoculate it in the liquid seed medium, culture it in a shake flask at 28°C for 36 hours, then insert 2% of the inoculum into the liquid seed medium, and culture it in a shake flask at 28°C 36h, as the seed solution for the next step of fermentation.

[0041] Fermentation medium: sucrose 17g / L, yeast extract 18g / L, KH 2 PO 4 0.5g / L, CaCO 3 0.2g / L, natural pH. The seed solution was inoculated in the fermentation medium with an inoculum amount of 5%, the fermentation temperature was 28° C., the rotation speed was 200 rpm, and the ventilation rate was 2 vvm. Start the membrane filter after 70 hours of fermentation, the membrane pore size is 0.5μm, and the operating pressure is 0.02MPa. The fermentation broth containing multi-antibiotics enters the separation device outside the fermentation t...

Embodiment 2

[0043] Seed medium: sucrose 13g / L, yeast extract 9g / L, KH 2 PO 4 0.3g / L, CaCO 30.1g / L. Pick a single colony of Streptomyces aureus chromogenicum NJYHYWG66302 from the plate medium and inoculate it in the liquid seed medium, culture it in a shake flask at 29°C for 40 hours, then insert 2% of the inoculum into the liquid seed medium, and culture it in a shake flask at 29°C 40h, as the seed solution for the next step of fermentation.

[0044] Fermentation medium: sucrose 18g / L, yeast extract 19g / L, KH 2 PO 4 0.6g / L, CaCO 3 0.2g / L, natural pH. The seed liquid was inoculated in the fermentation medium with an inoculum amount of 7%, the fermentation temperature was 29° C., the rotation speed was 220 rpm, and the ventilation rate was 3 vvm. Start the membrane filter after 75 hours of fermentation, the membrane pore size is 0.8μm, and the operating pressure is 0.01MPa. The fermentation broth containing multi-antibiotics enters the separation device outside the fermentation tank...

Embodiment 3

[0046] Seed medium: sucrose 15g / L, yeast extract 10g / L, KH 2 PO 4 0.4g / L, CaCO 3 0.2g / L. Pick a single colony of Streptomyces aureus chromogenicum NJYHYWG66302 from the plate medium and inoculate it in the liquid seed medium, culture it in a shake flask at 30°C for 48 hours, then insert 2% of the inoculum into the liquid seed medium, and culture it in a shake flask at 30°C 48h, as the seed solution for the next step of fermentation.

[0047] Fermentation medium: sucrose 19g / L, yeast extract 20g / L, KH 2 PO 4 0.7g / L, CaCO 3 0.3g / L, natural pH. The seed solution was inoculated in the fermentation medium with an inoculation amount of 8%, the fermentation temperature was 30° C., the rotation speed was 250 rpm, and the ventilation rate was 4 vvm. Start the membrane filter after 78 hours of fermentation, the membrane pore size is 1.0 μm, and the operating pressure is 0.01 MPa. The fermentation broth containing multi-antibiotics enters the separation device outside the fermenta...

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Abstract

The invention discloses a technology for realizing high-yield polyoxins by continuous fermentation coupled with separation. The technology includes inoculating the polyoxins producing strain seed liquid into a fermentation cylinder to perform fermental cultivation, starting a membrane filter in the fermentation cylinder when the fermentation is at later stage of the logarithmic phase, making the fermentation broth containing polyoxins go through the membrane filter and enter a separating device out of the fermentation cylinder, collecting the broth to obtain a fermentation broth A, intercepting the bacterial cell in the fermentation cylinder to perform fermentation, feeding nitrogen source medium into the fermentation cylinder while starting the membrane filter in the fermentation cylinder to maintain steady fermentation system in the fermentation cylinder, performing continuous fermentation to the end, collecting the broth to obtain a fermentation broth B, and performing refining and purifying processing of the fermentation broth A and fermentation broth B to obtain polyoxins. The technology releases the product inhibition in the fermentation, reduces the viscosity of the fermentation broth, shortens the fermentation period, and improves the product titer. Moreover, the technology couples continuous fermentation with separation with feeding device and separating device, improves the utilization rate of the device, and reduces the production cost.

Description

technical field [0001] The invention relates to a method for fermentation separation coupling control, which belongs to the field of bioengineering, and in particular to a process for realizing high-yield multi-antibiotics through continuous fermentation and separation coupling. Background technique [0002] Polyoxins, also known as pleiotropycin, polyoxin, and polyoxin, are secondary metabolites produced by Streptomyces cocoa var. Nucleoside agricultural antibiotics. The study found that polyoxins are composed of 14 structurally related components (PolyoxinsA-N). They have no fixed melting point and gradually decompose at around 200°C. They are easily soluble in water and insoluble in organic reagents such as acetone, ethanol and ether. It is stable under acidic conditions of pH 2.5-3.0, unstable under alkaline conditions, and stable to ultraviolet light. [0003] At present, plant diseases cause hundreds of billions of dollars in losses every year worldwide, and fungal ...

Claims

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Application Information

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IPC IPC(8): C12P21/02C07K1/34C12R1/465
CPCC12P21/02C07K1/34
Inventor 胡永红杨洋杨文革吴刚钱永根张棋曹翠翠程丽民马小平
Owner NANJING UNIV OF TECH
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