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Ethanol fermentation engineering bacteria capable of reducing fermentation byproduct

A technology of ethanol fermentation and engineering bacteria, applied in the directions of fermentation, microorganism-based methods, bacteria, etc., can solve problems such as affecting ethanol production, and achieve the effect of reducing the accumulation of by-products

Inactive Publication Date: 2010-11-03
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] However, it is known that in the fermentation process of six-carbon sugar and five-carbon sugar, many by-products of organic acid intermediate metabolism, such as formic acid and lactic acid, etc., are accumulated, and ethanol only accounts for a small part
The problem of by-products not only affects the yield of ethanol, but also has technical problems in the separation of by-products during subsequent treatment

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0025] Embodiment 1 Preparation of ethanol-resistant Escherichia coli ethanol fermentation engineering bacteria

[0026] A. Screening of Escherichia coli ethanol-resistant strain MG1655

[0027] Escherichia coli MG1655, JML09, BW25113, BL21, and BW25141 were cultured in LB medium, and after growing to the logarithmic phase, they were transferred to LB medium containing 4%, 6% ethanol for culture, (ethanol was filtered out through a 0.45 μm filter membrane) added to the LB medium after bacteria), 12h, 24h respectively measured OD in the liquid LB medium 600 The absorbance value was used to measure the difference in growth rate between screened strains. Compare OD 600 It was concluded that MG1655 was superior to other E. coli in ethanol tolerance.

[0028] B. Screening of mutant strains of ethanol-producing pyruvate formate lyase gene and lactate dehydrogenase gene of Escherichia coli

[0029] Escherichia coli MG1655 was made into competent cells by using calcium chloride, t...

Embodiment 2

[0133] Example 2 Engineering strain MPL-pzy507bc fermentation to produce ethanol

[0134] 1. Culture conditions

[0135] 1. 1M9 medium, the composition is:

[0136] Glucose 2%, Na 2 HPO 4 ·7H 2 O 64g, KH 2 PO 4 15g, NaCl 2.5g, NH 4 Cl 5.0g, 121°C, 15min. 1mol / L MgSO 4 2mL, 1mol / L CaCl 2 0.1mL, final volume 1L;

[0137] Xylose 10% Na 2 HPO 4 ·7H 2 O 64g, KH 2 PO4 15g, NaCl 2.5g, NH 4 Cl 5.0g, 121°C, 15min. 1mol / L MgSO 4 2mL, 1mol / L CaCl 2 0.1mL, final volume 1L;

[0138] 1.2 Culture and fermentation conditions

[0139] Inoculate MPL-pZY507bc into M9 medium and culture to OD 600 =0.2 or so, add 1.0mM IPTG to induce, when MPL-pZY507bc grows to OD 600 = 1.0, the bacteria were collected by centrifugation, inoculated into the fermentation medium described in 1.1, and made OD after inoculation 600 The value is about 1.0, 37 ℃ anaerobic culture.

[0140] 2. Determination of residual sugar in fermentation broth

[0141] High-performance liquid chromatography was...

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Abstract

The invention relates to an ethanol fermentation engineering bacteria capable of reducing a fermentation byproduct. The ethanol fermentation engineering bacteria is characterized in that a pyruvate formate-lyase gene and a lactate dehydrogenase gene of Escherichia coli are inactivated; and a Zymomonas mobilis gene is transferred into the Escherichia coli. When ethanol is produced by fermenting hexose and pentose with the engineering bacteria, lactic acid and formic acid are not produced and the byproduct is reduced, so that the ethanol productivity is improved.

Description

technical field [0001] The invention relates to an ethanol fermentation engineering bacterium capable of reducing fermentation by-products, and also relates to a preparation method of the engineering bacterium and its application in fermentation production of ethanol. Background technique [0002] Many plants, especially some plants that are often discarded, have high cellulose content. For example, in crop straw and residues, the content of cellulose and hemicellulose can account for more than 75% of the dry weight. If it can be used for fermentation to produce ethanol, it is a very good way to use it. [0003] Cellulose is mainly composed of six-carbon sugars and five-carbon sugars, such as glucose, xylose, arabinose, galactose, and mannose. [0004] E. coli contains all the necessary enzymes to utilize and ferment some six- and five-carbon sugars. However, the use of unmodified wild-type E. coli to ferment and produce ethanol has a very low yield and many by-products, w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/01C12N15/70C12P7/06C12R1/19
CPCY02E50/17Y02E50/10
Inventor 陈明马瑞强张维陆伟林敏徐玉泉
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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