Clostridium acetobutylicum strain and application thereof

A technology of Fusobacterium acetobutylicum and strains, applied in the directions of bacteria, microorganisms, biochemical equipment and methods, etc., can solve the problems of low butanol yield, low substrate conversion rate, etc., and achieves strong butanol tolerance and proportion The effect of high, total solvent yield

Inactive Publication Date: 2011-10-26
NANJING UNIV OF TECH
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  • Abstract
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Problems solved by technology

When its concentration reaches 13 g/L (Journal of Proteome Research2010, 9: 3046~3061), using Clostridium C. a

Method used

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  • Clostridium acetobutylicum strain and application thereof
  • Clostridium acetobutylicum strain and application thereof
  • Clostridium acetobutylicum strain and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0033] This embodiment illustrates that the present invention Clostridium acetobutylicum Screening method for BD518.

[0034] Clostridium acetobutylicum Clostridium acetobutylicum The XY16 original strain was activated and cultured at a culture temperature of 37°C, with a volume of 15-20 mL in a 50 mL Schott anaerobic bottle, filled with nitrogen for 2 minutes, and cultured for 12-18 hours to obtain a vigorously growing and robust bacterial liquid; Into a self-made continuous culture device with a liquid volume of 100-300 mL, the inoculum volume was 5%-15% (v / v), and after 12-18 hours of cultivation, a fresh medium containing butanol was added for domestication and selection. as attached figure 1 As shown, with the increasing butanol concentration, the OD of the strain decreased gradually, and the total acclimatization time was 744 h. Samples were taken at 472 h, 600 h, and 700 h of acclimatization time, spread on plates, and screened for mutants with high butanol toler...

Embodiment 2

[0051] This example illustrates Clostridium acetobutylicum Clostridium acetobutylicum BD 518 butanol resistance properties.

[0052] The medium formula described in this embodiment (% is mass percent):

[0053] Primary 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%, dipotassium hydrogen phosphate 0.1 %, ferrous sulfate heptahydrate 0.01%, the rest is water, pH 6.

[0054] Secondary 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%, dipotassium hydrogen phosphate 0.1 %, ferrous sulfate heptahydrate 0.01%, the rest is water, pH 6.

[0055] Butanol tolerance test medium: glucose 6%, ammonium acetate 0.22%, potassium dihydrogen phosphate 0.05%, dipotassium hydrogen phosphate 0.05%, sodium chloride 0.001%, magnes...

Embodiment 3

[0058] This example illustrates the passage stability of the mutant strain BD518.

[0059] In the fermentation medium with glucose as the carbon source, the passage stability of the mutant strain BD518 was detected, and the results of the passage fermentation test of the strain BD518 are shown in Table 2:

[0060] Table 2 strain BD518 passage fermentation test result

[0061]

[0062] 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 relates to a Clostridium acetobutylicum strain and application thereof, belonging to the technical field of biofermentation. The classification designation of the Clostridium acetobutylicum strain provided by the invention is Clostridium acetobutylicum BD518, and the preservation registration number is CCTCC NO:M2010308. The high-concentration butanol-tolerant strain BD518 is obtained by continuous culture, domestication and screening, and the butanol tolerance is up to 18g/L and is increased by 38.5% in comparison with the original strain (13g/L). When glucose is used as a carbon source, the total solvent and butanol yields in a 5L fermentation tank are respectively up to 21.1g/L and 13.2g/L and are respectively increased by 16.6% and 15.8% in comparison with the original strain. The Clostridium acetobutylicum strain has high butanol tolerance, high solvent yield and good repeatability, thereby having significant social meanings and economic values.

Description

technical field [0001] The invention relates to a strain of Fusobacterium acetobutylicum and its application, in particular to a Fusobacterium acetobutylicum strain resistant to high-concentration butanol and high solvent yield obtained through continuous cultivation, domestication and selection of a continuous culture device, and the fermentation of the strain The application in the butanol industry belongs to the technical field of biological fermentation. Background technique [0002] As a fuel, butanol has the advantages of high energy density, high stability to water, direct use in internal combustion engines, and convenient transportation. In today's increasingly severe energy crisis, butanol has broad development prospects as a fuel. Butanol is an important organic chemical raw material, widely used in paints, surface coatings, leather treatment, plastics and other fields. [0003] The production methods of butanol mainly include acetaldehyde condensation method, pro...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P7/16C12R1/145
Inventor 姜岷贺爱永郭亭杜腾飞陈可泉吴昊韦萍欧阳平凯
Owner NANJING UNIV OF TECH
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