A method for improving ethanol production efficiency by overexpressing the ino2 gene

A production efficiency, alcohol technology, applied in the field of bioenergy development, can solve the problems of difficult control inhibition, poison inhibition, etc., and achieve the effect of good industrial application value and prospects

Active Publication Date: 2018-10-16
JIANGNAN UNIV
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  • Claims
  • Application Information

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Problems solved by technology

In the past 10 years, the technology of producing ethanol by fermentation of thick mash has made great progress, but there is still a big gap compared with developed countries such as the United States. Osmotic pressure inhibition; toxicity inhibition caused by high concentration of ethanol in the later stage of fermentation; inhibition caused by uncontrollable high temperature during fermentation
At present, there is no literature report that the production efficiency of ethanol can be improved by overexpressing INO2

Method used

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  • A method for improving ethanol production efficiency by overexpressing the ino2 gene
  • A method for improving ethanol production efficiency by overexpressing the ino2 gene
  • A method for improving ethanol production efficiency by overexpressing the ino2 gene

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Effect test

Embodiment 1

[0033] Embodiment 1: Construction of Saccharomyces cerevisiae Genetic Engineering Bacteria

[0034] Using the ino4 single-gene deletion strain YOL108C purchased from Invotrogen Company and numbered Sc04163923_s1 as the starting strain, the overexpression is by constructing a high-expression plasmid pHAC181-INO2. stop codon), and then clone this fragment into the high-expression plasmid pHAC181, and finally carry out DNA sequencing on the correct recombinant, verify that the sequence does not mutate, and finally obtain the recombinant high-expression plasmid pHAC181-INO2, and then pass the lithium acetate method. pHAC181-INO2 was introduced into the ino4 single gene deletion strain to obtain a strain with high expression of INO2 gene.

Embodiment 2

[0035] Example 2: Saccharomyces cerevisiae alcoholic fermentation

[0036] Prepare YPD plate and YPD liquid medium in advance according to the preparation method of solid medium.

[0037]Streak activation of Saccharomyces cerevisiae genetically engineered bacteria of the present invention, wild-type strain BY4743BG (BY4743 containing G418 resistance gene) and Saccharomyces cerevisiae strain YOL108C with ino4 single gene deletion on YPD plate; ino4 single gene deletion strain YOL108C is in BY4743BG background obtained by replacing the gene INO4 with the G418 resistance gene in bacteria; (http: / / clones.lifetechnologies.com / cloneinfo.php?clone=yeast). The YPD plate was placed in a constant temperature incubator at 30°C for 2 days. Working in an ultra-clean bench, take a large single colony from each activated strain plate and inoculate it in 30 mL of YPD liquid medium, culture it on a shaker at 220 r / min at 30°C for 23 hours until saturated. After culturing for 23 hours, add 50...

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Abstract

The invention discloses a method for improving ethyl alcohol production efficiency through INO2 gene overexpression, and belongs to the technical field of biological energy development. According to the method, an engineering strain obtained by performing INO2 overexpression in a strain with single ino4 gene deletion is applied to fermentation production of ethyl alcohol; compared with a wild no-load contrast strain, when fermentation is performed for 52 h, the biomass is increased by 186.6%, the ethyl alcohol yield is increased by 204.1%, and the ethyl alcohol-thallus yield is increased by 5.6%. Thus, the method has great industrial application value and broad industrial application prospects, and meanwhile important information is provided for constructing excellent saccharomyces cerevisiae genetic engineering strains capable of achieving a high ethyl alcohol yield.

Description

technical field [0001] The invention relates to a method for improving alcohol production efficiency by overexpressing the INO2 gene, in particular to a method for improving the yield of ethanol to bacteria by using Saccharomyces cerevisiae with single gene deletion and overexpression of transcription factors, and belongs to the technical field of bioenergy development. Background technique [0002] Fossil energy (coal, oil, natural gas, etc.) is currently the most important energy in the world. With the development of economy and society in various countries in the world, the demand for energy is increasing day by day, but the reserves of fossil energy are limited; at the same time, fossil energy The global warming caused by the greenhouse gases produced by combustion and the resulting problems have always troubled people. Therefore, searching for and developing renewable new energy sources has become a hot spot of concern all over the world. Fuel alcohol has the advantage...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/19C12P7/06C12R1/185C12R1/865
CPCY02E50/10
Inventor 蒋伶活徐国强吴瑶杜婕
Owner JIANGNAN UNIV
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