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Clostridium beijerinckii with high stress resistance and application thereof

A technology of Clostridium beijerinckii and high resistance, applied in bacteria, microorganisms, biochemical equipment and methods, etc., to achieve the effects of high total solvent yield, great social and economic value, and high sugar conversion rate

Inactive Publication Date: 2011-09-07
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there are no patents and literature reports that can directly use undetoxified fiber acid hydrolysis sugar liquid to ferment butanol

Method used

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  • Clostridium beijerinckii with high stress resistance and application thereof
  • Clostridium beijerinckii with high stress resistance and application thereof
  • Clostridium beijerinckii with high stress resistance and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: This example illustrates the method of performing the first step of ion beam mutagenesis on the original strain of Clostridium beinii.

[0033] The original bacterial strain of Clostridium beinii NCIMB 8052 was purchased from the American Type Culture Collection (ATCC); the method for performing the first step of ion beam mutagenesis is as follows:

[0034] The original strain of Clostridium beinerii NCIMB 8052 was activated and cultivated at a culture temperature of 33-37°C, with a liquid volume of 10-15mL in a 25mL shaker bottle, filled with nitrogen for 3 minutes, and cultured for 12-18 hours to obtain vigorous growth and thick bacteria; Take freshly cultured cells and dilute to cell concentration OD 600 =0.05~0.1, spread on a sterilized empty petri dish, dry with sterile air; inject different doses with 10KeV, pulse each time for 5s, with an interval of 15s. After ion implantation mutagenesis, the bacterial film was eluted, and the survival rate was calc...

Embodiment 2

[0035] Embodiment 2: This example illustrates the method for further screening excellent Clostridium beinii.

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

[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%, the rest is water, pH 6.

[0038] (2) Resazurin plate medium: yeast powder 0.3%, peptone 0.5%, 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%, agar 1.5%, resazurin 0.002%, add 1 / 2 (v / v) of non-detoxified corn cob acid hydrolysis sugar solution, the rest is water, pH 6.

[0039] (3) Corn cob acid hydrolysis sugar solution plate medium: yeast powder 0.3%, peptone 0.5%, ammonium aceta...

Embodiment 3

[0053] Example 3: This example illustrates the passage stability of mutant strains IB4 and IB9.

[0054] In the fermentation medium with glucose as the carbon source, the passage stability of the mutant strains IB4 and IB9 was tested. The results of the subculture fermentation test of bacterial strain IB4 and IB9 are shown in Table 2:

[0055] Table 2 strain IB4 and IB9 subculture fermentation test results

[0056]

[0057] 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 Clostridium beijerinckii with high stress resistance which can directly utilize a virus-carrying wood fiber acid-hydrolyzed sugar solution for fermentation to produce butanol and an application thereof. The Clostridium beijerinckii disclosed by the invention has the category name of Clostridium beijerinckii IB4 and the collection number CCTCC No. is M2010310. Ion beam mutation is adopted, a resazurin flat plate containing inhibitors such as phenols and a shake flask are used for fermenting and screening, the obtained mutant strain uses a virus-carrying corncob acid-hydrolyzed sugar solution as a carbon source, the total solvent yield and butanol yield in a 2L fermentation tank are up to 10.3g / L and 7.1g / L respectively which are 4.3 times and 4.4 times of those obtained by adopting the starting strain cultivated under the same conditions, the sugar conversion rate is 0.35; and the Clostridium beijerinckii has the advantages of high stress resistance, high solvent yield, high sugar conversion rate and good repeatability, thereby being a good strain which is suitable for utilizing the wood fiber raw material to produce butanol.

Description

technical field [0001] The invention relates to a strain of Clostridium beinerii which can use non-detoxified lignocellulosic acid to hydrolyze sugar liquid and its application, which belongs to the technical field of biological fermentation. Background technique [0002] Butanol is an important C4 platform compound, which has been widely used in the manufacture of various fine chemicals; and as an important new biofuel with great potential, it has been valued. The United Nations International Energy Agency has listed bio-butanol as a Second generation biofuels. As a fuel, butanol has the advantages of high energy density, direct use in internal combustion engines, and convenient transportation. In today's increasingly severe energy crisis, butanol has broad development prospects as a fuel. Since domestic production cannot meet the demand, my country is already the world's largest importer of butanol. [0003] At present, butanol is mainly synthesized by chemical methods. ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P7/16C12R1/145
Inventor 姜岷郭亭汤艳奚永兰陈可泉李志刚孙佰军吴昊欧阳平凯
Owner NANJING UNIV OF TECH
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