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High ferulic acid stress resistance clostridium beijerinckii and application thereof

A technology of Clostridium beijerinckii and ferulic acid, which is applied in the direction of bacteria, microorganisms, biochemical equipment and methods, can solve the problems of production stoppage of bio-butanol manufacturers, and achieve high butanol production, strong stress resistance, and repeated good sex effect

Active Publication Date: 2014-03-26
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since 2008, my country has established many bio-butanol production lines with an annual production capacity of 100,000 tons; however, due to the continuous increase of starchy and molasses raw materials, most bio-butanol manufacturers are in a state of suspension

Method used

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  • High ferulic acid stress resistance clostridium beijerinckii and application thereof
  • High ferulic acid stress resistance clostridium beijerinckii and application thereof
  • High ferulic acid stress resistance clostridium beijerinckii and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] This example illustrates the first step of plasma mutagenesis of the original strain of Clostridium beijerinckii.

[0032] The method for the first step plasma mutagenesis of the original bacterial strain of Clostridium beijerinckii is as follows:

[0033] The original strain of Clostridium beijerinckii NCIMB8052 was activated and cultured at a culture temperature of 33-37°C, the volume of 50ml Schott anaerobic bottle was 15-20ml, filled with nitrogen for 3 minutes, and the culture time was 12-18 hours to obtain a vigorously growing bacterial liquid; take fresh Cultured cells were diluted to cell concentration OD 600 =1~1.5, drop on the sterilized and cooled slide, dry with sterile air; use helium as the discharge gas, 100W as the radio frequency power, 10SLM as the gas flow rate, and 10~1800s as the irradiation time Plasma mutagenesis was performed on the bacterial strain, and after the mutagenesis, the bacterial film on the carrier was eluted, and the survival rate w...

Embodiment 2

[0035] This example illustrates the method for screening good Clostridium beijerinckii.

[0036] Wherein, the medium formula used (% is mass percentage):

[0037] (1) Solid plate medium: yeast powder 0.3%, peptone 0.5%, soluble starch 1%, ammonium acetate 0.2%, sodium chloride 0.2%, magnesium sulfate heptahydrate 0.3%, potassium dihydrogen phosphate 0.1%, dihydrogen phosphate Potassium 0.1%, ferrous sulfate heptahydrate 0.01%, agar 1.5%, pH6.

[0038] (2) Resazurin plate medium: yeast powder 0.3%, peptone 0.5%, soluble starch 1%, ammonium acetate 0.2%, sodium chloride 0.2%, magnesium sulfate heptahydrate 0.3%, potassium dihydrogen phosphate 0.1%, phosphoric acid Dipotassium hydrogen 0.1%, ferrous sulfate heptahydrate 0.01%, agar 1.5%, resazurin 0.002%, ferulic acid 0.5%, pH6.

[0039] (3) Seed medium: yeast powder 0.3%, peptone 0.5%, soluble starch 1%, ammonium acetate 0.2%, sodium chloride 0.2%, magnesium sulfate heptahydrate 0.3%, potassium dihydrogen phosphate 0.1%, dipot...

Embodiment 3

[0050] This example illustrates the passage stability of the mutant strain M11.

[0051] In the fermentation medium with glucose as the carbon source, the passage stability of the mutant strain M11 was detected, and the results of the passage fermentation test of the strain M11 are shown in Table 2:

[0052] Table 2

[0053]

[0054] From the experimental results, it can be seen that after 7 consecutive passages, the total solvent production and butanol production of the two mutant strains are relatively stable, and they have good passage stability, and can be used as production strains for further research and development.

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Abstract

The invention discloses high ferulic acid stress resistance clostridium beijerinckii and an application thereof. The high ferulic acid stress resistance clostridium beijerinckii is classified and named Clostridium beijerinckii M11 with the collection number of CCTCC No:M2013423, and the collection date of the high ferulic acid stress resistance clostridium beijerinckii is December 14, 2013. A mutant strain obtained by adopting plasmas for mutating the clostridium beijerinckii and a ferulic acid containing resazurin flat plate and a shake flask for fermentation and screening can normally grow in a fermentation medium containing 0.8g / L of ferulic acid, the yield of butanol reaches 6.1g / L, and an original strain cultured under equal conditions hardly grows. Moreover, by using virus-carrying corncob acidolysis sugar liquid as a carbon source, the yield of butanol in a 2L fermentation tank reaches 6.4g / L, and the original strain cultured under equal conditions hardly grows. The high ferulic acid stress resistance clostridium beijerinckii has strong stress resistance, high butanol yield and good repeatability, and is an excellent strain suitable for the fermentation production of butanol by using wood fiber raw materials such as corncobs and the like.

Description

technical field [0001] The invention relates to a strain of Clostridium beijerinckii with high resistance to ferulic acid and its application in the acetone-butanol fermentation industry, belonging to the technical field of biological fermentation. Background technique [0002] As a new type of renewable liquid energy, butanol has been valued by more and more countries in the world; butanol has the advantages of high energy density, can be mixed with gasoline in any ratio, can be directly used in internal combustion engines, and is convenient for transportation. At present, the production method of butanol is mainly through chemical synthesis. With the increasingly serious energy crisis, the accelerated depletion of petroleum resources and the soaring price, butanol is produced by biological methods, as an effective and important platform compound and fuel for the production of biomass resources. The biotransformation technology has begun to be paid attention to again. Sinc...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P7/16C12R1/01
Inventor 应汉杰华莹郭亭刘俊王冬谢婧婧陈勇吴菁岚陈晓春朱晨杰庄伟
Owner NANJING UNIV OF TECH
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