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Clostridium acetobutylicum and construction method and purposes thereof

A clostridium acetobutylicum, construction method technology, applied in microorganism-based methods, biochemical equipment and methods, bacteria and other directions, can solve the problems of screening efficiency, quantity restricting industrial application, etc., and achieves wide applicability, short cycle, fast growing effect

Inactive Publication Date: 2010-09-29
NORTH CHINA PHARMA GROUP CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of this method is that this method still uses the traditional strain screening method, so its screening efficiency and quantity may seriously restrict the industrial application of this method.

Method used

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  • Clostridium acetobutylicum and construction method and purposes thereof
  • Clostridium acetobutylicum and construction method and purposes thereof
  • Clostridium acetobutylicum and construction method and purposes thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Construction of Clostridium acetobutylicum p1147 (CGMCC No3686)

[0044] a. Protoplast preparation: select isolated and cultivated ATCC824 bacteria, use lysozyme with a final concentration of 4 mg / ml to digest for 30 minutes, and heat the ATCC824 bacteria protoplasts to death at 55°C for 30 minutes.

[0045] b. Regeneration and Fusion:

[0046] Add the lethal protoplasts to 5ml PEG4000, induce fusion of the protoplasts for 5 minutes, collect by centrifugation for 10min, and add Ca at a final concentration of 60mM 2+ , and 75mMMg 2+ solution, mix the bacteria evenly, evenly spread on the SRA solid plate, and incubate at 37°C for 3 days in an anaerobic box to observe the results.

[0047] in:

[0048]ATCC824 was purchased from the market from the American Strain Collection.

[0049] Corn flour medium: Each liter of medium contains 50g of corn flour.

[0050] RCM medium: Each liter medium contains 3.0g of yeast powder, 10.0g of peptone, 10.0g of beef extract, 5.0g of ...

Embodiment 2

[0058] Construction of Clostridium acetobutylicum p1147 (CGMCC No3686)

[0059] a. Protoplast preparation:

[0060] Collect ATCC824 bacteria, use lysozyme with a final concentration of 10 mg / ml to digest for 40 minutes, and heat the protoplasts to death at 60°C for 40 minutes.

[0061] b. Add the lethal protoplasts to 10ml PEG6000, induce fusion of the protoplasts for 10min, collect by centrifugation for 10min, and add Ca with a final concentration of 50mM 2+ , and 75mMMg 2+ solution, mix the bacteria evenly, evenly spread on the SRA solid plate, in an anaerobic box, incubate at 37°C for 2 days and observe the results.

[0062] in:

[0063] ATCC824 was purchased from the market from the American Strain Collection.

[0064] Corn flour medium: Each liter of medium contains 50g of corn flour.

[0065] RCM medium: Each liter medium contains 3.0g of yeast powder, 10.0g of peptone, 10.0g of beef extract, 5.0g of glucose, 10.0g of starch, 3.0g of sodium acetate, 0.5g of L-cystei...

Embodiment 3

[0070] Experimental comparison of Clostridium acetobutylicum p1147 constructed and cultured in Example 1 with commercially available ATCC824 for the production of mixed alcohols.

[0071] 1. Clostridium acetobutylicum p1147 cultured in Example 1 (for experimental group 1) and commercially available ATCC824 (for control group) were fermented with 6% corn fermented glutinous rice medium at a temperature of 37° C. for 48-96 hours.

[0072] 2. Detection of fermentation products

[0073] Take the fermentation broth and use GC-7890A (Agilengt Technologies) to detect the changes of fermentation components, detection conditions: chromatographic column DB-624 (30m×0.32mm×1.8μm, Agilengt Technologies); detector: FID (300°C); carrier gas: N 2 ;Temperature mode: 1-stage temperature program, initial temperature 55°C, keep 8min, rate 45°C / min, final temperature 190°C, keep 9min; split ratio: 30:1; sampling method: autosampler (Agilengt Technologies), Injection volume: 0.2 μL; quantitative ...

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Abstract

The invention discloses a clostridium acetobutylicum p1147 (CGMCCNo3686) which is prepared by the protoplast fusion of clostridium acetobutylicum ATCC 824. The strain can be used for fermenting and producing polydiol which contains isopropanol. The strain provided by the invention has the advantages of rapid growth, high reproduction, short cycle and stability, and the polydiol fermented and produced by the strain has high yield and good quality.

Description

technical field [0001] The invention relates to Clostridium acetobutylicum and its construction method and application. Background technique [0002] In recent years, due to the increasing scarcity of fossil resources such as petroleum, the conversion of biomass into chemicals and liquid fuels by acetone-butanol fermentation has become the focus of attention again. The traditional acetone-butanol fermentation takes corn or molasses as the substrate, inserts Clostridium acetobutylicum (Clostridium acetobutylicum) for fermentation, and its fermentation products are acetone, butanol, and ethanol, and its ratio is 3:6:1, Mixed alcohols account for 70% of the total dosage, of which butanol accounts for 60% of the total dosage. Because of its high energy content, low volatility, wide tolerance to water when mixed with gasoline, high safety compared with other biofuels, and low greenhouse gas emissions, it is generally believed that increasing the proportion of butanol in the solv...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N15/03C12P7/16C12P7/06C12P7/04C12R1/145
CPCY02E50/17Y02E50/10
Inventor 刘力强牛昆贾娟娟杨明武芳闫海荣范俊辉邸胜苗冯文亮杨丽萍时蕾石晨光王正品
Owner NORTH CHINA PHARMA GROUP CORP
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