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Clostridium acetobutylicum strain and screening method and application thereof

A technology of Fusobacterium acetobutylicum and acetone butanol, applied in the direction of microorganism-based methods, biochemical equipment and methods, bacteria, etc., to achieve significant social significance and economic value

Inactive Publication Date: 2012-09-12
NANJING TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are only reports on the screening of acetone-butanol producing bacteria by 2-deoxy-D-glucose plate at home and abroad

Method used

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  • Clostridium acetobutylicum strain and screening method and application thereof
  • Clostridium acetobutylicum strain and screening method and application thereof
  • Clostridium acetobutylicum strain and screening method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] This example illustrates the method of performing the first step of ion beam mutagenesis on the original strain of Fusobacterium acetobutylicum.

[0032] The original strain of Fusobacterium acetobutylicum came from self-screening, the specific method is as follows:

[0033] 1. Use the fermentation screening medium containing 25g / L butanol to enrich the butanol-resistant microorganisms in the soil samples in Nanjing area, and screen out the strains that can tolerate 25g / L butanol. Streak the bacterial solution in the sample tube capable of growth on the solid plate medium, culture at 37°C for 48 hours, and obtain a single colony.

[0034] Among them, fermentation screening medium: butanol 1%, yeast powder 0.2%, peptone 0.3%, soluble starch 1%, ammonium acetate 0.2%, sodium chloride 0.2%, magnesium sulfate 0.3%, potassium dihydrogen phosphate 0.1%, phosphoric acid Dipotassium hydrogen dipotassium 0.1%, ferrous sulfate heptahydrate 0.01%, pH6.

[0035] 2. Pick the singl...

Embodiment 2

[0040] This example illustrates the method for further screening of excellent Fusobacterium acetobutylicum.

[0041] Wherein, the medium formula (% is mass percent) used:

[0042] (1) Solid plate medium: yeast powder 0.2%, peptone 0.3%, soluble starch 1%, ammonium acetate 0.2%, sodium chloride 0.2%, magnesium sulfate 0.3%, potassium dihydrogen phosphate 0.1%, dipotassium hydrogen phosphate 0.1 %, 0.01% ferrous sulfate 7 hydrate, pH6.

[0043] (2) Xylose plate medium: corn steep liquor 0.1% (solid content is 40-50%), xylose 1%, ammonium acetate 0.2%, sodium chloride 0.2%, magnesium sulfate 0.3%, potassium dihydrogen phosphate 0.1 %, dipotassium hydrogen phosphate 0.1%, ferrous sulfate 7 hydrate 0.01%, agar 1.5%, pH 5-7.

[0044] (3) 2-deoxy-D-glucose plate medium: yeast powder 0.2%, peptone 0.3%, soluble starch 1%, ammonium acetate 0.2%, sodium chloride 0.2%, magnesium sulfate 0.3%, potassium dihydrogen phosphate 0.1% , dipotassium hydrogen phosphate 0.1%, ferrous sulfate he...

Embodiment 3

[0065] This example illustrates the passage stability of mutant strains XY16 and XY10.

[0066] The results of the subculture fermentation test of bacterial strain XY16 and XY10 are shown in Table 2:

[0067] Table 2

[0068]

[0069] 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 capable of quickly generating butanol by fermenting xylose and starch, and a screening method and application thereof. The invention discloses Clostridium acetobutylicum, which is classified and named Clostridium acetobutylicumXY16 with the preservation and registration number of CCTCC NO:M2010011. The strain obtained by ion beam mutation and screening of a xylose plate, a 2-deoxy-D-glucose plate, a bromocresol green plate and a resazurin plate can quickly and efficiently generate the butanol by fermenting xylose and starch, has the advantages of high yield of total solvent, high butanol ratio and good repeatability, and is a good strain suitable for industrial production.

Description

technical field [0001] The invention relates to a Fusobacterium acetobutylicum strain with high solvent yield and high butanol ratio obtained by ion beam mutagenesis, a screening method of the strain and its application in the solvent fermentation industry, belonging to biological fermentation technology field. Background technique [0002] Butanol is an important organic chemical raw material, widely used in paints, surface coatings, leather treatment, plastics and other fields. 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. The apparent consumption of butanol in my country reached 680,000 tons in 2005, and it is estimated that the domestic demand for butanol in 2010 will reach 972,000 tons. Since domestic production cannot meet the demand, my country is already th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N13/00C12N15/01C12P7/16C12R1/145
Inventor 何冰芳欧阳平凯姜岷靳孝庆吴斌柏中中
Owner NANJING TECH UNIV
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