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Method for producing ethyl alcohol through crop stalks by use of recombinant zymomonas mobilis

A technology for Zymomonas and ethanol production, applied in the field of microbial fermentation, can solve problems such as narrow substrate utilization range, and achieve the effects of solving the narrow substrate utilization range, high activity and yield, and improving efficiency

Inactive Publication Date: 2016-03-02
HUBEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, Z.mobills also has its own limitations, the substrate utilization range is narrow, and only glucose, fructose and sucrose can be utilized

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0027] The preparation of the activated seed solution in step (2) is to inoculate 2.5ml of Aspergillus niger spore suspension and 2.5ml of Trichoderma reesei spore suspension in a 250ml shake flask respectively, the total liquid volume is 50ml, and the fermentation conditions are: temperature 28°C, rotation speed 150rpm, fermentation time 7 days. And the fermentation broth was mixed 1:1.

[0028] The total volume of the enzymolysis solution in step (3) is 50ml. The enzymolysis conditions are: temperature 50°C, enzymolysis time 4d.

[0029] In the present invention, the crude enzyme liquid of cellulase fermented by Trichoderma reesei and the crude enzyme liquid of cellulase fermented by Aspergillus niger are mixed at 1:1, and the crude enzyme liquid after mixing is 12:1 according to the ratio of material to liquid , that is, adding 1 g of straw to 12 ml of crude enzyme solution, and enzymatically hydrolyzing it for 4 days at 50°C. The recombinant Zymomonas mobilis was inocul...

Embodiment 1

[0032] 1. Materials

[0033] (1) Strains: Trichoderma reesei: T.reesei RUTC30, Aspergillus niger: Aspergillus niger, Zymomonas mobilis: Z. mobilis

[0034] (2) PDA solid medium: PDA is potato dextrose agar medium. Wash and peel the potatoes, weigh 200g of potatoes and cut them into small pieces, add them to a beaker, add water and cook for 30min until the potatoes can be gently pierced by a glass . Filter with gauze, add 20g of glucose and 20g of agar, and dilute to 1L. Autoclave at 115 for 30 minutes, pour solid plate.

[0035] (3) Seed medium: straw 10g / L pretreated by ammonia explosion, bran 5g / L, yeast extract 5g / L, tryptone 3g / L, Tween-80 40 drops / L, Mandles trace element salt Solution 1ml / L, Mandles nutrient salt concentrated solution 100ml / L, pH is 4.8 and the concentration is 50ml / L of citric acid buffer solution of 1mol / L. Autoclave at 115 for 30 minutes and set aside.

[0036] (4) Fermentation medium: dry straw 30g / L pretreated by ammonia explosion, bran 25g / L,...

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Abstract

The invention provides a method for producing ethyl alcohol through crop stalks by use of recombinant zymomonas mobilis. According to the method, the crop stalks are used as a base material, aspergillus niger and trichoderma are mixed and fermented to produce a crude enzyme solution containing cellulase, the crop stalks are processed through the crude enzyme solution to enable cellulose in the crop stalks to be decomposed to produce reducing sugar, and finally recombinant zymomonas mobilis is used for fermenting the enzymatically-decomposed reducing sugar to produce ethyl alcohol. In the method, fermentation products of aspergillus niger and trichoderma are mixed according to the ratio of 1: 1 to obtain the cellulase crude enzyme solution, so that the product feedback inhibition is effectively controlled, and the cellulase crude enzyme with higher activity and yield is obtained. As zymomonas mobilis is inoculated in the enzymatically-decomposed fermentation liquor containing reducing sugar, the ethyl alcohol production efficiency through the crop stalks by use of zymomonas mobilis is greatly improved, and defects that the use range of a zymomonas mobilis substrate is narrow, and the crop stalks cannot be effectively utilized are overcome. According to the method, the conversion rate of ethyl alcohol through the crop stalks is 5%, that is, 100 g of the crop stalks can produce 5 g of ethyl alcohol.

Description

technical field [0001] The invention belongs to the technical field of microbial fermentation, and in particular relates to a method for producing ethanol from straw with recombinant Zymomonas mobilis. Background technique [0002] Energy conversion of straw is a utilization method of straw under the new situation. Some other countries in the world have tried the exploration of straw energy earlier. Denmark is one of the first countries in the world to use straw to generate electricity. In the United States, part of the straw is utilized through more traditional methods; part of the straw is used as raw material to produce fuel ethanol through fermentation to replace traditional liquid energy. At the same time, Japan and Canada have begun research on straw energy, and have made progress in using straw to extract alcohol. [0003] The main component of straw is cellulose, and the cellulose must be degraded in order to use straw. Cellulase is not a single enzyme, but a gen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/10C12R1/01
CPCC12P7/10C12N1/205C12R2001/01Y02E50/10
Inventor 邓颖
Owner HUBEI UNIV OF TECH
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