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Method for preparing bioethanol

A bio-ethanol and ethanol technology, applied in biochemical equipment and methods, microorganism-based methods, microorganisms, etc., can solve the problems of waste of xylose, slow growth rate, and inability to utilize xylose, so as to improve utilization efficiency and increase production. The effect of high rate and yield

Inactive Publication Date: 2012-09-12
JIANGSU UNIV OF SCI & TECH
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Problems solved by technology

Since the commonly used microorganisms such as Saccharomyces cerevisiae and Zymomonas mobilis can only use six-carbon sugars such as glucose to produce ethanol, they cannot use xylose produced by plant fiber degradation, resulting in waste of xylose produced by plant fiber degradation, resulting in low ethanol yields. not high; and these microbial ethanol-producing bacteria cannot directly use plant cellulose to produce bioethanol
[0005] At present, it has been reported that genetically engineered yeast, Escherichia coli and other engineered bacteria directly use cellulose to produce ethanol, but genetically engineered bacteria usually require antibiotics to maintain their metabolic capabilities, and have slow growth, slow metabolism, low ethanol yield, Can not efficiently utilize natural plant fibers and other shortcomings, so genetically engineered bacteria show shortcomings in the direct use of plant fibers to produce ethanol

Method used

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  • Method for preparing bioethanol

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Embodiment 1

[0025] The determination of embodiment 1 fermentation medium

[0026] According to the culture characteristics of cellulolytic microorganisms and oil-producing microorganisms, the formula of the mixed-bacteria fermentation medium was designed by using single-factor experiments and orthogonal experiments. According to the needs of microbial growth and metabolism, the fermentation medium is screened and designed as follows:

[0027] (1) Medium Ⅰ: 9g Na 2 HPO 4 12H 2 O, 1.5g KH 2 PO 4 , 1.2 mg 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 2O, 0.0016g MnSO 4 ·H 2 O, 0.0014g ZnSO 4 ·H 2 O, 0.002g CoCl 2 , 1000mL H 2 O; pH is 4-6;

[0028] (2) Medium II: 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; pH is 4-6;

[0029] (3) Medium III: 1g peptone...

Embodiment 2

[0031] Embodiment 2 Fermentation condition investigation

[0032] (1) Activation of strains

[0033] Preparation of seed solution: Aspergillus niger (Aspergillus niger, DSMZ 821) was cultured in PDA medium at 25-30°C for 3-5 days at constant temperature

[0034] The ethanol-producing bacteria Candida shehatae (Candida shehatae, ATCC34887) was cultured in a YEPD medium at a constant temperature of 25-30°C for 3-5 days.

[0035] (2) mixed bacteria fermentation ethanol method

[0036] Respectively from the mixed ratio of strains (Aspergillus niger: Candida shohata, the biomass ratio is 1:0.5, 1:1 or 1:2 respectively.), the total inoculation amount, fermentation temperature and initial pH value of fermentation to ethanol obtained The influence of the rate is investigated to determine the relevant factors and their scope of influence. Based on the single factor experiment, the optimal level of the mixing ratio of strains, total inoculum, fermentation temperature and fermentation...

Embodiment 3

[0042] The preparation of embodiment 3 bioethanol

[0043] (1) Preparation of seed liquid: Aspergillus niger (DSMZ 821), which produces cellulase, was cultured at constant temperature in PDA medium to prepare Aspergillus niger seed liquid, and Candida shehatae (Candida shehatae, ATCC34887) The seed liquid of Candida shohata was obtained by constant temperature cultivation in YEPD medium, the cultivation temperature was 25-30° C., and the cultivation time was 3 days.

[0044] (2) Plant fibers such as straw and Aspergillus niger seed liquid were cultured in the medium (I) as described in Example 1 with constant temperature and shaking for 5 days, and the volume of Aspergillus niger seed liquid was 2% of the volume of the medium (I) ; Insert Candida shohata seed liquid, the biomass ratio of Aspergillus niger seed liquid and Candida shohata seed liquid is 1:0.5, continue constant temperature shaking culture for 3 days, culture temperature is 28 ℃, shaking speed 150r / min;

[0045...

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Abstract

The invention provides a method for preparing bioethanol. The method comprises the following steps of: performing constant-temperature shaking culture on plant fiber and seed liquid of a cellulase producing strain for 0 to 5 days in a fermentation culture medium, inoculating seed liquid of an ethanol producing strain into the fermentation culture medium, and continuing to perform constant-temperature shaking culture for 3 to 7 days; and filtering fermentation liquor, and distilling a filtrate under reduced pressure to obtain the bioethanol. The method for preparing the bioethanol is high in yield and low in cost, is environment-friendly, and is suitable for industrialized production.

Description

technical field [0001] The invention relates to a method for preparing bioethanol, in particular to a method for preparing bioethanol by using plant fibers through microbial mixed fermentation. Background technique [0002] With the rapid development of world industry and the gradual depletion of petroleum and other fossil resources, the research of using renewable resources to produce bioenergy has attracted more and more attention from researchers at home and abroad. Cellulose is the most abundant renewable resource on the earth. It has the advantages of being cheap, degradable and non-polluting to the ecological environment. Therefore, the production of fuel ethanol from lignocellulose to replace petroleum fuel is of great significance to the sustainable development of society and economy. [0003] However, cellulose is a polymer compound with a linear structure connected by glucose groups through glycosidic bonds. Its structure is complex, and there are a large number of...

Claims

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Application Information

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IPC IPC(8): C12P7/14C12R1/685C12R1/72C12R1/885C12R1/84
Inventor 李强季更生吴再强谷绪顶万富费娟娟
Owner JIANGSU UNIV OF SCI & TECH
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