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Clostridium beijerinckii and method for preparing biological butanol through fermentation of xylose residue serving as raw material thereof

A technology of Clostridium beijerinckii and xylose residues, applied in microorganism-based methods, biochemical equipment and methods, fermentation and other directions to improve sugar production and solvent production, solve bacterial strain capacity and raw material shortages, and reduce inhibitors Effect

Active Publication Date: 2012-10-10
QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] The object of the present invention is to provide a high-yielding Clostridium beijerinckii strain, so that its anaerobic fermentation butanol output can be improved, so as to solve the problem of the ability of biological fermentation to produce butanol strains

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  • Clostridium beijerinckii and method for preparing biological butanol through fermentation of xylose residue serving as raw material thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0050] Example 1: This example illustrates the method of subjecting the original strain Clostridium beijerinckii NCIMB 8052 to ethyl methanesulfonate (EMS) mutagenesis.

[0051] The original strain Clostridium beijerinckii C. beijerinckii NCIMB 8052 was purchased from the British National Collection of Industrial, Marine and Food Cultures (NCIMB), and the method for EMS mutagenesis was as follows:

[0052] The original strain Clostridium beijerinckii NCIMB 8052 was activated and cultured at 33-37°C for 12-18 hours to obtain vigorous growth and thick bacteria liquid; take 1 mL of fresh cultured bacteria liquid, centrifuge to collect the bacteria, and use Wash the cells with 50mM sterile phosphate buffer (pH 7.0) for 3 times, then add to the seed medium containing 0.5-2% EMS, treat for 10-60 minutes, discard the supernatant by centrifugation, and add 5% sodium thiosulfate The reaction was terminated; by calculating the survival rate, 1% EMS was the optimal mutagenic dose, and 20...

Embodiment example 2

[0053] Implementation Case 2: The present invention illustrates the method for screening excellent Clostridium beijerinckii.

[0054] Wherein, the medium formula used is as follows:

[0055] (1) Starch plate medium: soluble starch 5g / L, glucose 5g / L, yeast powder 1g / L, dipotassium hydrogen phosphate 0.5g / L, potassium dihydrogen phosphate 0.5g / L, ammonium acetate 2.2g / L, Magnesium sulfate heptahydrate 0.2g / L, manganese sulfate monohydrate 0.01g / L, ferrous sulfate heptahydrate 0.01g / L, sodium chloride 0.01g / L, p-aminobenzoic acid 0.001g / L, vitamin B 1 0.001g / L, biotin 0.0001g / L, agar 15g / L, the rest is water, pH 6.5.

[0056] (2) 2-deoxy-D-glucose plate medium: soluble starch 5g / L, 2-deoxy-D-glucose 5g / L, yeast powder 1g / L, dipotassium hydrogen phosphate 0.5g / L, potassium dihydrogen phosphate 0.5g / L, ammonium acetate 2.2g / L, magnesium sulfate heptahydrate 0.2g / L, manganese sulfate monohydrate 0.01g / L, ferrous sulfate heptahydrate 0.01g / L, sodium chloride 0.01g / L, p-amino Ben...

Embodiment 4

[0069] Example 4: This example illustrates the passage stability of the mutagenized strain Y-3.

[0070] In the fermentation medium with glucose as the carbon source, the subculture stability of the mutant strain Y-3 was tested. The results showed that after 8 transfers, the amylase activity was stable, and the butanol and total solvent production were stable, which was consistent with the shake flask fermentation screening results. unanimous.

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Abstract

Disclosed is a clostridium beijerinckii capable of producing butanol with high yield. The classification and nomenclature of the clostridium beijerinckii is Clostridium beijerinckii Y-3, with an accession number of CGMCC 5805. The method for preparing biological butanol through fermentation of xylose residue serving as raw material in the invention comprises the steps of performing calcium hydroxide detoxification pretreatment, carrying out anaerobic hydrolysis, preparing C. beijerinckii Y-3 anaerobic fermentation medium and adopting fermentation to produce biological butanol. According to the invention, the high-yielding butanol bacterial strain screened by using ethyl methyl sulfonate (EMS) mutation, starch plate and 2-deoxy-D-glucose plate is capable of producing butanol by using xylose residue treated by calcium hydroxide detoxification process as raw material and through fermentation. In this way, the problems of inadequate strain ability and raw material shortage during the process of producing butanol through traditional biological fermentation are solved and the transformation from industrial waste to high value-added biological energy is realized. The method is an environment-friendly method for preparing biological butanol and has a wide prospect of industrial application.

Description

technical field [0001] The invention belongs to the technical field of biological fermentation, and in particular relates to Clostridium beijerinckii Y-3 with high butanol yield and a method of utilizing industrial waste xylose slag through calcium hydroxide detoxification pretreatment, anaerobic enzymatic hydrolysis and Clostridium fermentation Process for producing biobutanol. Background technique [0002] As an important organic chemical raw material, butanol is widely used in pharmaceutical industry, plastic industry, organic industry, printing and dyeing, etc. In addition to being used as a solvent, butanol is also a potential bulk power fuel, its combustion value is equivalent to that of gasoline, and it is a substitute for automobile oil. The production process of butanol mainly includes chemical synthesis method and microbial fermentation method. [0003] With the depletion of petroleum resources, it is difficult to produce butanol by propylene oxo synthesis using ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P7/16C12R1/145
Inventor 李福利张婉璐刘自勇
Owner QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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