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Method of producing ethanol by fermenting immobilized yeast cells taking waste molasses as raw material

An immobilized yeast and cell fermentation technology, applied in the field of new immobilized yeast cells, can solve the problems of low ethanol yield, increase cost, affect the activity of free cells, etc., achieve low-cost utilization, increase service life, and improve ethanol Yield effect

Active Publication Date: 2017-05-31
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But, find that when the fermentative production of ethanol is with waste molasses as carbon source, because also contain other organic acids in molasses, colloid, ash and other components can seriously affect the activity of free cells, so that ethanol yield is not high, in order to achieve better The production effect requires pretreatment of acid and alkali processes before the use of molasses, which increases the cost of production

Method used

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  • Method of producing ethanol by fermenting immobilized yeast cells taking waste molasses as raw material
  • Method of producing ethanol by fermenting immobilized yeast cells taking waste molasses as raw material
  • Method of producing ethanol by fermenting immobilized yeast cells taking waste molasses as raw material

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Experimental program
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Effect test

Embodiment 1

[0032] The immobilization of embodiment 1 yeast cell

[0033]3 bottles of seed culture medium (250mL Erlenmeyer flask, liquid volume 120mL): molasses 50g / L (calculated based on the amount of glucose contained in molasses), CaCl 2 0.022g, (NH 4 ) 2 SO 4 0.72g, (NH 4 ) 2 HPO 4 0.36g, MgSO 4 0.036g, yeast powder 1.8g, H 2 O270mL, molasses needs to be sterilized separately and then mixed with other sterilized medium components.

[0034] Preparation of composite immobilized carrier: 300mL of mixed solution of 6% (w / w) PEG and 4% (w / w) SA and 1% (w / w) activated carbon (18 grams of PEG in 300mL mixed solution, 12g of SA , 3g of activated carbon); 5% (w / w) of CaCl 2 300mL of the solution was sterilized at 121°C for 15min respectively.

[0035] Inoculate the strains on the slant of the test tube into the seed medium, culture at 30°C and 200r / min for 20 hours, centrifuge at 6000 rpm for 1min to collect the bacteria, wash the bacteria 3-5 times with sterile normal saline, a...

Embodiment 2

[0036] The optimization experiment of embodiment 2 composite immobilization carrier

[0037] In order to find out the best immobilization conditions, the concentration of the immobilization materials sodium alginate (SA), polyethylene glycol (PEG) and activated carbon was optimized through three factors and three levels of orthogonal experiments, and assuming the relationship between each factor There is no interaction between them.

[0038] According to Example 1, the yeast cells were immobilized and cultured on a shaking table at 200 rpm at 30°C, and the molasses ethanol fermentation experiment was carried out according to the ratio of the volume of the immobilized cells to the volume of the fermentation broth of 1:7, wherein the volume of the fermentation broth was 750ml, temperature 30°C, rotation speed 80rpm, adjust pH to 5.0 with ammonia water, ventilate, record the highest ethanol concentration that can be achieved in the fermentation broth.

[0039] Table 1: Factor le...

Embodiment 3

[0044] The control experiment of embodiment 3 free bacteria fermentation and different immobilized carrier fermentation

[0045] In the experiment, after the yeast cells were immobilized according to the method of Example 1, the value-added culture was carried out on a shaker at 200 rpm at 30°C, and the seed culture of free bacteria was used as a control experiment. After 20 hours of value-added culture, the volume of immobilized cells and the fermentation broth The ratio of the volume ratio is 1:7 to the fermentation medium, and the free bacteria are inoculated into the fermentation medium with the same amount of bacteria. The volume of the fermentation broth is 750ml, the temperature is 30°C, the rotation speed is 80rpm, the ammonia water is adjusted to PH5.0, aeration is performed, and samples are taken at regular intervals to measure the production of ethanol and the consumption of sugar. The sugar here is calculated by the total reducing sugar. . Experimental results suc...

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Abstract

The invention discloses a method of producing ethanol by fermenting immobilized yeast cells taking waste molasses as a raw material. The method comprises the following steps: diluting cultivated yeast cells to a bacterial suspension, and uniformly mixing the bacterial suspension with an immobilized carrier to prepare immobilized yeast cells, wherein the immobilized carrier is a mixed solution of sodium alginate (SA) and polyethylene glycol (PEG) and activated charcoal; and performing enrichment culture on the prepared immobilized yeast cells, then transferring the culture to a fermenting culture medium, and producing ethanol by fermenting by taking waste molasses as the raw material. The method disclosed by the invention is suitable for fermentation in a system of molasses which is not treated, so that the output of ethanol is improved, and the carrier loss rate is reduced; the highest ethanol concentration after fermentation for 58h can reach 68g / L, the carrier is not damaged after batched fermentation for 90h, and the capacity of producing ethanol is not declined after continuous fermentation for near 400 hours. The method is simple to operate and low in cost, and provides a good referential method for fermenting the immobilized cells in the system of molasses.

Description

technical field [0001] The invention belongs to the technical field of biological fermentation, and in particular relates to a novel immobilized yeast cell used for fermentation in molasses raw materials without any pretreatment. Background technique [0002] As an important industrial raw material, ethanol is widely used in food, chemical industry, medicine, dyestuff, national defense and other fields. At the same time, it is also a renewable clean energy, and its combustion value as a fuel reaches 26900kJ / kg. [0003] As one of the by-products of the sugar industry, molasses is one of the by-products of the sugar industry. Because its main component is sugar, in order to improve the utilization of waste molasses biomass resources, using molasses as a carbon source in the fermentation process has been extensively studied. In addition, molasses also contains high mineral ion Na + 、K + , Mg 2+ , Ca 2+ , Cl - etc., also contains a small amount of protein, nitrate, amides...

Claims

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Application Information

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IPC IPC(8): C12P7/08C12N11/08C12N11/10C12N11/14C12R1/865
CPCC12N11/08C12N11/10C12N11/14C12P7/08
Inventor 陈可泉李梅何珣张阿磊齐雁斌魏国光欧阳平凯
Owner NANJING UNIV OF TECH
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