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Method for sectional regulation and control for continuous strain culture

A technology for cultivating strains and segmented regulation, applied in microorganism-based methods, biochemical equipment and methods, bacteria, etc., can solve problems such as restricting downstream applications, fluctuations in fermentation broth components, and changes in bacterial activity. The effect of improving the utilization rate of raw materials, simplifying the production process, and high batch stability

Pending Publication Date: 2019-03-22
WANHUA CHEM GRP CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Patent CN107735495 uses a semi-continuous process for fermentation, but adding a large proportion of fresh medium will cause large fluctuations in the components of the fermentation broth, which in turn will lead to changes in the activity of the bacteria
[0006] To sum up, the current production of rhamnolipids generally adopts batch fermentation method, and the production efficiency is low, resulting in high cost, which seriously limits its application in downstream fields.

Method used

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  • Method for sectional regulation and control for continuous strain culture

Examples

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Effect test

Embodiment 1

[0037] Using Pseudomonas aeruginosa as the strain, it was inoculated into three 3L fermenters connected in series after activation and shake flask culture, with a liquid volume of 70%. The fermentation medium contains glycerol 60g / L, yeast extract 10g / L, potassium dihydrogen sulfate 1g / L, disodium hydrogen phosphate 1g / L, ferrous sulfate 0.2g / L, magnesium sulfate 0.1g / L, and calcium chloride 0.1g / L, sodium chloride 0.1g / L, defoamer 1%. The fermentation temperature was controlled to 35°C, the pH of the fermentation broth was 7, the aeration volume of the fermentation broth was 1.0 vvm, and the stirring speed was 300 rpm, and stage I fermentation was carried out. After 20h, enter the second stage, and continuously add fresh feed medium to the 1# fermentor at a rate of 8mL / L / h. The glycerol content in the medium is increased to 72g / L, other components remain unchanged, and the aeration rate is 2.0 vvm, stirring speed 500rpm, fed fermentation for a total of 10h; then enters stage ...

Embodiment 2

[0039] Using Pseudomonas putida as the strain, it was inoculated into three 3L fermenters connected in series after activation and shake flask culture, with a liquid volume of 70%. The fermentation medium contains glucose 20g / L, rapeseed oil 20g / L, peptone 7g / L, sodium nitrate 8g / L, potassium dihydrogen sulfate 1g / L, disodium hydrogen phosphate 1g / L, ferrous sulfate 0.2g / L, magnesium sulfate 0.1g / L, calcium chloride 0.1g / L, sodium chloride 0.1g / L, defoamer 3%. The fermentation temperature was controlled to 30°C, the pH of the fermentation broth was 8, the aeration rate of the fermentation broth was 2.0 vvm, and the stirring speed was 600 rpm, and stage I fermentation was carried out. After 50h, enter the second stage, and continuously add fresh feed medium to the 1# fermentor at a rate of 16mL / L / h. The glucose and rapeseed oil content in the medium is increased to 30g / L and 30g / L, respectively. Other components remain unchanged, the aeration volume is 3.0vvm, the stirring spe...

Embodiment 3

[0041] Using Pseudomonas aeruginosa as the strain, it was inoculated into three 7L fermenters connected in series after activation and shake flask culture, with a liquid volume of 70%. The fermentation medium contains glycerol 30g / L, cottonseed oil 50g / L, yeast powder 2g / L, ammonium sulfate 3g / L, potassium dihydrogen sulfate 1g / L, disodium hydrogen phosphate 1g / L, and ferrous sulfate 0.2g / L, magnesium sulfate 0.1g / L, calcium chloride 0.1g / L, sodium chloride 0.1g / L, defoamer 2%. The fermentation temperature was controlled to 32°C, the pH of the fermentation broth was 7.5, the aeration volume of the fermentation broth was 1.5 vvm, and the stirring speed was 500 rpm, and stage I fermentation was carried out. After 30h, enter the second stage, and continuously add fresh feed medium to the 1# fermentor at a rate of 12mL / L / h. The glycerin and cottonseed oil content in the medium are increased to 54g / L and 90g / L, respectively. The composition is unchanged, the aeration rate is 2.5vv...

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Abstract

The invention discloses a method for sectional regulation and control for continuous strain culture. By adopting the method, nutrient substances are continuously replenished into a fermentation liquidto maintain strain metabolism, and at the same time, the fermentation liquid is continuously led out and is fed into a serial fermentation tank for further fermentation; to increase the strain metabolism velocity, parameters such as dissolved oxygen, pH values and replenished material constitution are respectively regulated and controlled at different stages. By adopting the method, the times ofstrain culture and equipment cleaning are reduced, the production process can be simplified, and the fermentation cost can be reduced. In addition, due to sectional regulation and control on the fermentation process, the yield of rhamnolipid can be increased, the fermentation production efficiency can be improved, the product batch stability can be improved, and the method is easily applied to industrial production.

Description

Technical field [0001] The invention relates to a method for continuously cultivating bacterial species with stepwise regulation, in particular to a method for continuous culturing of Pseudomonas aeruginosa or Pseudomonas putida, and its application in the fermentation production of rhamnolipids . Background technique [0002] There is a huge amount of surfactants worldwide, and they are widely used in many fields such as petroleum, environment, agriculture, daily chemicals, food and medicine. However, most of the surfactants on the market are synthesized from petroleum-based chemicals, which are limited by the reserves and non-renewable characteristics of petroleum resources. Therefore, the development of biosurfactants has attracted wide attention in recent years. [0003] In addition to excellent surface activity, biosurfactants also have the characteristics of green, environmental protection and sustainability, and can be used in many fields such as petroleum, environment, ag...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P19/44C12R1/385C12R1/40
CPCC12N1/20C12P19/44
Inventor 王竞辉陈长生姜西娟张雅萍石森黄真真孔令晓鞠昌迅黎源华卫琦
Owner WANHUA CHEM GRP CO LTD
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