Controllable high-efficiency microorganism growth medium/liquid and preparation method thereof

A high-efficiency microorganism and culture medium technology, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems such as the inability to guarantee product quality, the inability to guarantee stable and uniform distribution of dissolved oxygen, and the inability to guarantee product quality stability. , to achieve the effect of optimizing product quality

Inactive Publication Date: 2009-03-11
陈祥海
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method cannot guarantee the stability of the dissolved oxygen level and the uniform distribution of oxygen in the overall environme

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Embodiment 1: aerobic Gram-positive bacteria fermentation broth

[0016] Prepare 15 liters of peptone broth according to the following formula: 10 g / L peptone (DIFCO), 3 g / L beef extract (Sigma), 5 g / L sodium chloride. Place in a 30 liter fermenter, add 10 6 beta-strep, the fermenter was rotated at 50 rpm at 37°C. After culturing for 24 hours, 10 ml of perfluorotripropylamine with a purity greater than 99% and 20 ml of perfluorobutane were added respectively. Continue culturing at the same rotational speed. The rate of passing oxygen was 8 ml / min. Dynamically observed dissolved oxygen level was 18mg / 100ml, with an average fluctuation of less than 3% per minute. After bacterial fermentation for 144 hours, the temperature was raised to 65°C and kept for two hours, then taken out and cooled to 4°C for storage. The concentration of bacterial DNA quantified was 3.4±0.2 ng / ml. The same method was repeated 3 times, and the average error of bacterial DNA concentration bet...

Embodiment 2

[0018] Embodiment 2: moderately aerobic Gram-negative bacteria fermentation broth

[0019] Prepare 15 liters of peptone broth according to the following formula: 10 g / L peptone (DIFCO), 3 g / L beef extract (Sigma), 5 g / L sodium chloride, pH 6.8-7.6. Place in a 30 liter fermenter, add 10 6 R. marcescens, the fermenter was rotated at 50 rpm at 25 °C. After culturing for 24 hours, 10 ml of perfluorotripropylamine with a purity greater than 99% was added respectively. Continue culturing at the same rotational speed. The rate of passing carbon dioxide is 3 ml / min. Dynamically observe the dissolved oxygen level at 5 mg / 100 ml, with an average fluctuation of less than 2% per minute. After 48 hours of bacterial fermentation, the temperature was raised to 65°C and kept for two hours, then taken out and cooled to 4°C for storage. The concentration of bacterial DNA quantified was 5.6±0.04 ng / ml. The same method was repeated 3 times, and the average error of bacterial DNA concentrati...

Embodiment 3

[0021] Embodiment 3: Type A Streptococcus fermentation broth

[0022] Prepare 15 liters of peptone broth according to the following formula: 10 g / L peptone (DIFCO), 3 g / L beef extract (Sigma), 5 g / L sodium chloride, pH 6.8-7.6. Place in a 30 liter fermenter, add 10 6 R. marcescens, the fermenter was rotated at 50 rpm at 25 °C. After culturing for 24 hours, 5 ml of perfluorotripropylamine with a purity greater than 99% was added respectively. Continue culturing at the same rotational speed. The rate of oxygen flow was 3 ml / min. Dynamically observed dissolved oxygen level was 18mg / 100ml, with an average fluctuation of less than 5% per minute. After bacterial fermentation for 96 hours, raise the temperature to 65°C for two hours, take it out and cool it to 4°C for storage. The concentration of bacterial DNA quantified was 4.7±0.4 ng / ml. The same method was repeated 3 times, and the average error of bacterial DNA concentration between batches was less than 5%. Endotoxin con...

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Abstract

The invention discloses a method for preparing a controllable highly-efficient microbial culture medium/liquid, which is suitable for the biotechnology industry such as microbial pharmacy and fermentation and other industries which need different microbial culture media/liquids. The controllable highly-efficient microbial culture medium/liquid is characterized in that the culture medium/liquid is dissolved with a dissolved oxygen control agent of fluorocarbon added according to a volume ratio between 1 to 10 and 1 to 10000 and also dissolved with a dissolved oxygen control agent of oxygen, carbon dioxide or helium, wherein the flow of oxygen is 1 to 10 ml/min, the flow of carbon dioxide is 1 to 5 ml/min or the flow of helium is 1 to 5 ml/min flow. The microbial culture medium/liquid can be used for cells amplification, is applicable to the preparation of bacterial immune agents, the beer manufacture, the biological purification and other bioengineering manufacture industries; the fluorocarbon in the microbial culture medium/liquid can be removed by heating or freeze-drying processes.

Description

(1) Technical field [0001] The invention relates to a method for preparing a controllable and high-efficiency microbial culture medium / liquid, which is suitable for use in biological industries such as microbial biopharmaceuticals, fermentation, and other industries that require different microbial culture medium / liquid requirements. (2) Background technology [0002] The growth environment of microorganisms is the key to determining the growth and metabolism of microorganisms, so the preparation of appropriate medium / solution is the core process step that affects the quality of bioengineering products. Growth time, temperature, pH, dissolved oxygen content and other working conditions are the main factors affecting the growth environment of microorganisms, among which the dissolved oxygen content is the most difficult to control. Different microorganisms have different requirements for the level of dissolved oxygen in the culture medium / solution. For example, some Gram-posi...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/01
Inventor 陈祥海
Owner 陈祥海
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