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Three-stage genetic transcription control method for improving cellulosic ethanol yield and genetic engineering bacterial strain

A cellulosic ethanol and gene transcription technology, applied in the field of genetic engineering strains, three-stage gene transcription regulation to improve cellulosic ethanol production, and can solve problems affecting ethanol production and yield, low consumption rate, low conversion rate, etc.

Inactive Publication Date: 2013-06-12
CAPITAL NORMAL UNIVERSITY
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Problems solved by technology

[0005] However, the plasmid burden, metabolic burden, and various environmental stress responses brought about by gene transcription regulation technology in the fermentation and metabolism of genetic engineering may affect the final ethanol production and yield, which is the uncertainty of this research technology
At the same time, the low consumption rate and low conversion rate of xylose utilization, although researchers have tried different promoters, different copy numbers, different background sources of microbial strains, and different genetic protein engineering, have not changed the current face of cellulosic ethanol technology. bottlenecks and difficulties

Method used

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  • Three-stage genetic transcription control method for improving cellulosic ethanol yield and genetic engineering bacterial strain
  • Three-stage genetic transcription control method for improving cellulosic ethanol yield and genetic engineering bacterial strain
  • Three-stage genetic transcription control method for improving cellulosic ethanol yield and genetic engineering bacterial strain

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Embodiment 1

[0044] 1. Construction of mixed sugar co-fermentation strain

[0045] The starting strain WT comes from the industrial strain of Saccharomyces cerevisiae produced by Angel Yeast Company. After splitting the spores from industrial diploids and transforming them into haploids, the dominant strains with better fermentation characteristics were screened through fermentation, and they were used as the original industrial starting strains for our experiment.

[0046] The plasmid p61 was transformed into WT to obtain WXY1, and its strain contained two genes pADH1-RPE1 and pPGK1-XDH; the plasmid p62 was transformed into WXY1 to obtain WXY2, and the strain additionally contained pADH1-XR4m and pPGK1-XK; WXY2, to get WXY3, this strain additionally contains multiple copies of pPGK1-RKI1 and pADH1-TAL1, and this strain can initially utilize xylose to produce ethanol; transform the plasmid pUC-3XK270R into WT, and get WXY4, this strain contains three genes pADH1- XR(K270R) / pPGK1-XDH / pPGK1-...

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Abstract

The invention relates to the field of biology preparation of ethanol, in particular to a three-stage genetic transcription control method for improving cellulosic ethanol yield and genetic engineering bacterial strain. The method comprises a step of expressing genes of XR, XDH, XK, RPE1, RKI1 and TAL1 in saccharomyces cerevisiae by building of expression plasmids of the genes of XR, XDH, XK, RPE1, RKI1 AND TAL1, wherein KGD1 is used for starting sub-mediation rate-limiting gene XK, and HSP26 is used for starting sub-mediation key rate-limiting gene TAL1. The cellulose alcoholic fermentation process is divided into three stages including an anaerobic glucose ferment stage, an aerobic xylose respiratory metabolism stage and a ferment later stage period which has a heat shock characteristic and gives priority to high temperature restraint. A target gene is enabled to be high efficiently expressed in each stage so that cellulose zymolyte can be high efficiently transferred to cellulosic ethanol.

Description

technical field [0001] The invention relates to the field of biological preparation of ethanol, in particular to a method for improving cellulose ethanol production through three-stage gene transcription regulation and a genetically engineered bacterial strain. Background technique [0002] With the continuous progress of human civilization, global energy issues have become prominent and have become an important factor restricting the sustainable and stable development of the global economy. Fuel ethanol is a clean and renewable biomass energy. Cellulose fuel ethanol technology has a wide range of raw materials and is a sustainable and environmentally friendly technology. The construction of its core technology system has become a global energy strategy. land. After lignocellulose is decomposed, it is mainly converted into glucose and xylose, which are fermented by Saccharomyces cerevisiae under anaerobic conditions to produce ethanol. The present invention aims to use lig...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/81C12N1/19C12P7/10C12R1/865
CPCY02E50/10
Inventor 萧伟曹利民汤兴良田雪蕾
Owner CAPITAL NORMAL UNIVERSITY
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