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Using Ammonium Ion to Regulate Lincomycin Fermentation Process and Increase Lincomycin Yield

A technology of lincomycin and ammonium ions, applied in the fields of fermentation, microbiology, biochemical equipment and methods, etc., can solve the problem of not being able to feed back the dynamic adjustment information of the fermentation process in time, not fully utilizing the production potential of Streptomyces lincomycin, and unclear biosynthetic routes And other issues

Active Publication Date: 2011-12-21
EAST CHINA UNIV OF SCI & TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Lincomycin (Lincomycin) belongs to the lincosamine antibiotics. The fermentation process of Streptomyces lincolnensis (S.lincoinensis) to produce lincomycin is very complicated, and its biosynthetic route is still unclear. way
[0003] At present, there are only studies on the factors affecting the fermentation process of lincomycin on the shake flask scale, but due to the limitations of mixing and sampling, shake flask fermentation can neither fully develop the production potential of Streptomyces lincomycin, nor can it feed back the fermentation in time process dynamic adjustment information
However, there is no report on the factors affecting the fermentation of lincomycin in a large-scale bioreactor.

Method used

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  • Using Ammonium Ion to Regulate Lincomycin Fermentation Process and Increase Lincomycin Yield
  • Using Ammonium Ion to Regulate Lincomycin Fermentation Process and Increase Lincomycin Yield

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 150L

[0064] Embodiment 150L fermenter cultivation

[0065] The inoculum size is 20%; the air volume (VVM) per minute per unit volume (L) of the fermentation broth is 1.4L; the stirring speed is 300-550r / min; the culture temperature is 30°C; the tank pressure is 0.05MPa; the initial culture fluid volume is 30L ; Culture period 184h.

[0066] test methods

[0067] Dry cell weight (DCW): Take 10ml of fermentation broth, weigh it wet, filter it with Buchner funnel, wash the wet solid three times with water, dry at 80°C for 12 hours, weigh it, and calculate DCW (g / L).

[0068] Lincomycin Potency: Chinese Pharmacopoeia (2005 Edition) Part Two Regulations.

[0069] NH 4 + Content: Determination of ammonium ion by phenol-hypochlorite reaction [see: Xu Chendong et al., Discussion on the method of determination of ammonium ion by phenol-hypochlorite reaction [J]; Journal of Wuxi Light Industry University: Food and Biotechnology, 1998, 17(1):34-38].

[0070] Amino nitrogen (NH 2 -N) co...

Embodiment 2

[0072] Example 2NH 4 + Dynamic effects on nitrogen source metabolism

[0073] The inventors have identified four typical supplemental ammonium sulfate regimens (Table 1) to investigate NH 4 + effect on the fermentation process. In order to investigate the NH in the basal medium 4 + Changes in the fermentation process when the content is higher, the content of ammonium sulfate in the F1 tank basal medium is set to 7g / L (equivalent to 103.139mmol / L); For the dynamic change of the process, the feeding process of F3 tank was designed; in order to investigate the dynamic change of the fermentation process in the early stage of fermentation and when the rate of ammonium sulfate feeding was low, the feeding process of F4 tank was designed. where [NH 2 -N], check every 2 hours.

[0074] Feeding process scheme of table 1 ammonium sulfate

[0075] tank number

0h [NH 4 + ]

Ammonium Sulfate Supplementation Protocol

F1

103.139mmol / L

[NH 2 -N...

Embodiment 3

[0084] Example 3NH 4 + Dynamic effects on glucose metabolism

[0085] According to the measurement, at 0 hour, the total sugar content of the 4 tanks of culture fluid were 6.3, 6.52, 6.32 and 6.78g / 100ml respectively. Before 16h, the total sugar consumption rate of tanks F2, F3 and F4 changed basically the same. After 16 hours, ammonium sulfate was supplemented according to Table 1: both tanks F3 and F4 began to replenish ammonium sulfate for the first time at 16 hours, and the rate of ammonium sulfate supplementation in the former was lower than that in the former. The former is larger, resulting in the former's total sugar consumption rate being greater than the latter; the F2 tank begins to add ammonium sulfate at 20h, and at 16-20h, its NH 4 + Concentrations were lower than tanks F3 and F4, and the rate of total sugar consumption was lower ( figure 2 ), indicating that NH 4 + Concentration is positively correlated with enzyme activity of glucose degrading enzyme.

...

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Abstract

The invention belongs to the field of microbial fermentation and discloses an optimized lincomycin fermentation production method. The method comprises adjusting the ammonium ion concentration in the initial fermentation medium to 57±5mmol / L, cultivating Streptomyces lincomycin; When the concentration of ammonium ions is 0.2-1.5mmol / L, add ammonium ions, and then control the concentration of ammonium ions in the fermentation medium to 2-5mmol / L until the end of fermentation. For the first time, the present inventor has studied the influencing factors of Streptomyces lincomycin fermentative production of lincomycin on a larger scale, and summarized an optimized lincomycin fermentation production method, which can greatly increase the yield of lincomycin .

Description

technical field [0001] The invention belongs to the field of microbial fermentation, and more specifically, the invention relates to an optimized lincomycin fermentation production method. Background technique [0002] Lincomycin (Lincomycin) belongs to the lincosamine antibiotics. The fermentation process of Streptomyces lincolnensis (S.lincoinensis) to produce lincomycin is very complicated, and its biosynthetic route is still unclear. way. [0003] At present, there are only studies on the factors affecting the fermentation process of lincomycin on the shake flask scale, but due to the limitations of mixing and sampling, shake flask fermentation can neither fully develop the production potential of Streptomyces lincomycin, nor can it feed back the fermentation in time Dynamic adjustment information of the process. However, there is no report on the research on the factors affecting the fermentation of lincomycin in a large-scale bioreactor. Contents of the invention ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P17/16C12R1/565
Inventor 储炬李啸张嗣良庄英萍钟承赞葛友群杭海峰王永红郭美锦夏建业章付林杨明
Owner EAST CHINA UNIV OF SCI & TECH
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