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Method for producing butanol by utilizing co-culture fermentation of thermoanaerobacterium thermosaccharolyticum and clostridium acetobutylicum

A technology of Clostridium acetobutylicum and anaerobic bacteria, applied in the field of microbial fermentation, can solve the problem of high production cost, and achieve the effects of increasing butanol yield and reducing cost

Active Publication Date: 2018-03-06
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Purpose of the invention: In order to solve the problem that the traditional butanol fermentation method uses grain or other starchy by-products as raw materials, the production cost is too high. butanol method

Method used

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  • Method for producing butanol by utilizing co-culture fermentation of thermoanaerobacterium thermosaccharolyticum and clostridium acetobutylicum
  • Method for producing butanol by utilizing co-culture fermentation of thermoanaerobacterium thermosaccharolyticum and clostridium acetobutylicum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Using xylose as a carbon source to produce butanol using Clostridium acetobutylicum ATCC824

[0026] Pick a single colony of the strain Clostridium acetobutylicum ATCC 824 from the plate and inoculate it into 5mL fermentation medium, cultivate it at 37°C and 150rpm for 48h, and then inoculate it into a new fermentation medium with an inoculation amount of 5% v / v. Shake culture at 37°C and 150rpm, adjust the pH to 5.5 every 12h, measure the concentration of various products with GC after 120h, and the final butanol production is 8.97g / L.

[0027] The above fermentation medium formula is 1g / L NaCl, 0.75g / L K 2 HPO 4 , 0.75g / L KH 2 PO 4 , 3g / L yeast powder, 0.5g / L MgCl 2 ·6H 2 O, 0.3g / L NH 4 Cl, 0.015g / L CaCl 2 2H 2 O, 1.5g / L FeCl 2 4H 2 O, 0.3g / L KCl, xylose 60g / L, adjust the pH to 5.5, blow nitrogen for 10-20min, and sterilize at 121°C for 15min.

Embodiment 2

[0028] The impact of embodiment 2 different xylan concentrations on final butanol output

[0029] (1) Inoculate the pyrolytic sugar hyperthermia anaerobic bacteria into the activation medium with an inoculum size of 5% v / v, activate at 55°C and 120rpm for 60h, and adjust the pH to 7.5 every 12h;

[0030] The activation medium formula is: 1g / L NaCl, 0.75g / L K 2 HPO 4 , 0.75g / L KH 2 PO 4 , 3g / L yeast powder, 0.5g / L MgCl 2 ·6H 2 O, 0.3g / L NH 4 Cl, 0.015g / L CaCl 2 2H 2 O, 1.5g / L FeCl 2 4H 2 O, 0.3g / LKCl, xylan 60g / L, the solvent is water, adjust the pH to 7.5;

[0031] (2) Inoculate the activated thermosaccharolytic thermoanaerobe M5 into the fermentation medium with an inoculation amount of 5% v / v, ferment at 55° C. and 120 rpm for 60 hours, and adjust the pH to 7.5 every 12 hours to obtain a fermentation broth;

[0032] The formula of the fermentation medium is: 1g / L NaCl, 0.75g / L K 2 HPO 4 , 0.75g / L KH 2 PO 4 , 3g / L yeast powder, 0.5g / L MgCl 2 ·6H 2 O,0.3g / L NH...

Embodiment 3

[0037] Example 3 Effect of different inoculation methods of Clostridium acetobutylicum ATCC 824 on the yield of butanol

[0038] The method is the same as in Example 2, except that the concentration of xylan in the fermentation medium is 60g / L, and the culture solution of Clostridium acetobutylicum activated in step (3) is put into a centrifuge at 4°C and centrifuged at 6000rpm for 10min , then the supernatant is poured out, and the bacterium slime is added in the fermented liquid of step (2), wherein the volume of the Clostridium acetobutylicum cultured liquid that has been activated is the same as the volume of the fermented liquid; Then continue to cultivate at 37°C, with a rotating speed of 150rpm, Adjust the pH to 5.5 every 12h and ferment for 132h;

[0039] During the cultivation process, samples were taken every 12 hours and the butanol production was measured. When co-cultured with the inoculation method of Example 2, the butanol output was only 2.61g / L; when co-culti...

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Abstract

The invention discloses a method for producing butanol by utilizing co-culture fermentation of thermoanaerobacterium thermosaccharolyticum and clostridium acetobutylicum. The method comprises the following steps: (1) inoculating activated thermoanaerobacterium thermosaccharolyticum into a xylan-containing fermentation medium to be fermented so as to obtain fermentation liquor; and (2) inoculatingactivated clostridium acetobutylicum into the fermentation liquor obtained in the step (1), and fermenting to produce the butanol. When the thermoanaerobacterium thermosaccharolyticum M5 is cultured for 60 hours, the clostridium acetobutylicum bacterial sludge is added, and then the butanol yield is the highest and reaches 8.34 g / L, which is the highest butanol yield obtained by co-culture by taking the xylan as a unique carbon source at present. According to the method disclosed by the invention, the industrial production cost of the butanol is reduced, and the problem that hemicellulose canbe utilized by the thermoanaerobacterium thermosaccharolyticum while the butanol yield is low is solved. The method disclosed by the invention has important application value.

Description

technical field [0001] The invention belongs to the field of microbial fermentation and relates to butanol fermentation, in particular to a method for producing butanol by co-cultivating and fermenting pyrolytic sugar high-temperature anaerobic bacteria and Clostridium acetobutylicum. 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. The production methods of butanol mainly include acetaldehyde condensation method, propylene carbonyl synthesis method and fermentation method. With the soaring price of petroleum and the accelerated depletion of resources, the production of butanol by fermentation has received extensive attention, and has gradually become one of the research hotspots of bioenergy. [0003] The traditional fermentation method t...

Claims

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Application Information

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IPC IPC(8): C12P39/00C12P7/16C12R1/01C12R1/145
CPCC12P7/16C12P39/00Y02E50/10
Inventor 信丰学蒋羽佳姜岷陆家声董维亮章文明方艳马江锋
Owner NANJING UNIV OF TECH
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