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Method for preparing biodiesel by directly using cellulose through mixed fermentation

A mixed fermentation and biodiesel technology, applied in fermentation, biofuels, etc., can solve the problems that cannot be used to produce biodiesel, high-yield microorganisms can not use cellulose, etc., to achieve rich raw materials, solve energy shortages, and rapid regeneration cycle Effect

Inactive Publication Date: 2015-03-04
JIANGSU UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0006] Technical problem to be solved: the present invention aims at the technical problem that cellulase-producing bacteria can efficiently degrade cellulose materials, but cannot be used to produce biodiesel and other products; and microbial oil-producing bacteria cannot utilize cellulose, and provides a cellulose Method for producing biodiesel by mixed fermentation of degrading bacteria and oil-producing microorganisms

Method used

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  • Method for preparing biodiesel by directly using cellulose through mixed fermentation

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Determination of medium for mixed fermentation:

[0038] According to the culture characteristics of cellulolytic microorganisms and oleaginous microorganisms, the medium components described in step (1) were optimized by using single factor experiments, orthogonal experiments, response surface analysis and other methods, and the mixed fermentation medium was screened and designed as follows :

[0039] Medium Ⅰ: 10g cellulose; 9g Na 2 HPO 4 12H 2 O; 1.5 g KH 2 PO 4 ; 1.2mg FeNH 4 -Citrate; 0.34g (NH 4 ) 2 SO 4 ;0.15g Peptone【Peptone】; 0.15g Yeast extract; 0.3g CaCl 2 , 0.3g MgSO 4 , 0.005gFeSO 4 ·7H 2 O; 0.0016g MnSO 4 ·H 2 O; 0.0014g ZnSO 4 ·H 2 O; 0.002g CoCl 2 ; 1000mL H 2 O, the pH is adjusted to a weakly acidic pH of 6.0-6.8.

[0040] Medium Ⅱ: 10g cellulose; 2g KH 2 PO 4 ; 1.4g (NH 4 ) 2 SO 4 ;0.3g MgSO 4 ; 0.3g CaCl 2 ; 0.5g peptone; 0.005g FeSO 4 ·7H 2 O; 0.0016g MnSO 4 ·H 2 O, 0.0014g ZnSO 4 ·H 2 O; 0.002g CoCl 2 , 1000mLH 2 O;...

Embodiment 2

[0058] The oil-producing microorganism Rhodococcus opacus (Rhodococcus opacus ACCC41043) and the cellulose-degrading bacteria Trichoderma reesei (Trichoderma reesei ATCC 26921) were used as mixed strains for mixed fermentation. Mixed fermentation medium adopts medium Ⅰ: 15g cellulose; 9g Na 2 HPO 4 12H 2 O; 1.5 g KH 2 PO 4 ; 1.2mg FeNH 4 -Citrate; 0.34g (NH 4 ) 2 SO 4 ; 0.15g Peptone; 0.15g Yeast extract; 0.3g CaCl 2 , 0.3g MgSO 4 , 0.005g FeSO 4 ·7H 2 O; 0.0016gMnSO 4 ·H 2 O; 0.0014g ZnSO 4 ·H 2 O; 0.002g CoCl 2 ; 1000mL H 2 O, the pH was adjusted to pH 6.8. The fermenter was a 5L fermenter, and the culture temperature was 28°C. Trichoderma reesei was inoculated according to 2% v / v inoculum. After 12 hours of cultivation, Rhodococcus turbide was inoculated into the fermenter according to 2% v / v inoculum for mixed bacteria fermentation.

[0059] Extraction of fatty acids: Ferment for 5 days, take 2L of the bacterial liquid, centrifuge to get the precipitate, ...

Embodiment 3

[0064] The oil-producing microorganism Rhodococcus opacus (Rhodococcus opacus ACCC41043) and the cellulose-degrading bacteria Aspergillus niger (Aspergillus niger DSMZ 821) were used as mixed strains for mixed fermentation. Medium Ⅱ: 18g bagasse; 2g KH 2 PO 4 ; 1.4g (NH 4 ) 2 SO 4 ;0.3g MgSO 4 ; 0.3g CaCl 2 ; 0.5g Peptone; 0.005g FeSO 4 ·7H 2 O; 0.0016gMnSO 4 ·H 2 O, 0.0014g ZnSO 4 ·H 2 O; 0.002g CoCl 2 , 1000mL H 2 O; the pH was adjusted to pH 6.8; the fermentation vessel was a 250mL Erlenmeyer flask, and the culture temperature was 28°C. Aspergillus niger was inoculated according to 2% v / v inoculum, and Rhodococcus turbide was inoculated according to 2% v / v inoculum after 12 hours of cultivation Carry out mixed fermentation.

[0065] Extraction of fatty acids: Ferment for 5 days, take 50mL of the bacterial liquid, centrifuge to get the precipitate, and extract the precipitate with 500mL extractant (volume ratio: chloroform:methanol=2:1) ​​overnight. Centrifuge...

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Abstract

The invention discloses a method for preparing biodiesel by directly using cellulose through mixed fermentation. The mixed fermentation is performed by using a plant fiber degrading strain and a grease producing strain, and ethanol is produced by directly using natural plant fiber such as straws. The method comprises the following steps of: designing a culture medium for the mixed fermentation by optimization methods such as a response surface method, and optimizing to obtain the fermentation conditions such as the optimum temperature of mixed fermentation, optimum mixing proportion of strains and optimum mixing time of the strains by utilizing the methods such as orthogonal test and response surface methodology; according to the optimized fermentation conditions, by using the plant fiber as a carbon source, degrading the cellulose into fermentable sugar by using a cellulase producing strain, and adding grease-producing microorganisms into the culture system for the mixed fermentation; and by using the sugar which is produced by the constant enzymolysis of the plant fiber as the carbon source, fermenting to produce grease by using the grease producing strain, and performing methyl esterification reaction of the microbial grease to obtain the biodiesel.

Description

technical field [0001] The invention belongs to the technical field of bioenergy and biocatalysis, and in particular relates to a microbial mixed fermentation method for producing biodiesel. Background technique [0002] Biodiesel refers to a renewable diesel fuel that can replace petrochemical diesel oil, which is made from oil crops, wild oil plants, engineering microalgae and other aquatic vegetable oils, animal oils, and catering waste oils through transesterification processes. Biodiesel is a kind of biomass energy, which is a mono-alkyl ester of long-chain fatty acid obtained from biomass by pyrolysis and other technologies. Biodiesel is a mixture of complex organic components with extremely high oxygen content. These mixtures are mainly organic compounds with large molecular weight, including almost all kinds of oxygen-containing organic compounds, such as: ethers, esters, aldehydes, ketones, phenols, organic acids, alcohols Wait. Biodiesel has the following excelle...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P39/00C12P7/64
CPCY02E50/13Y02E50/10
Inventor 李强季更生谷绪顶费娟娟唐玉斌
Owner JIANGSU UNIV OF SCI & TECH
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