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Reorganized saccharomyces cerevisia bacterial strain and preparation method and application thereof

A technology of yeast strain and Saccharomyces cerevisiae, which is applied in the fields of genetic engineering and metabolic engineering, can solve the problem that the utilization rate of arabinose needs to be improved, and achieves the improvement of the utilization rate of raw materials, the fermentation efficiency, the ethanol yield, and the high sugar alcohol conversion rate. Effect

Active Publication Date: 2019-04-12
JILIN COFCO BIOCHEM +2
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Problems solved by technology

However, the patent only discloses the replacement of Bacillus licheniformis L-arabinose isomerase (araA), Escherichia coli L-ribulokinase (araB) and L-ribulone with preferred codons from Candida The codon of sugar-5-phosphate 4-epimerase (araD) and the technical scheme of introducing it into Candida, and the utilization rate of arabinose still needs to be improved

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  • Reorganized saccharomyces cerevisia bacterial strain and preparation method and application thereof
  • Reorganized saccharomyces cerevisia bacterial strain and preparation method and application thereof
  • Reorganized saccharomyces cerevisia bacterial strain and preparation method and application thereof

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Embodiment approach

[0055] According to a preferred embodiment of the present invention, the recombinant yeast strain is a recombinant yeast strain (S.C araABD) deposited under the accession number CGMCC No. 16830.

[0056] The preparation method of the recombinant yeast strain provided by the third aspect of the present invention, the method comprises: introducing the gene expression cassette comprising the genes of the following (1) to (3) into the yeast strain:

[0057] (1) araA gene from Pediococcus lactis,

[0058] (2) araB gene from Lactobacillus plantarum,

[0059] (3) araD gene from Lactobacillus plantarum.

[0060] According to a preferred embodiment of the present invention, the nucleotide sequence of the araA gene from Pediococcus lactis is shown in SEQ ID NO: 1; the nucleotide sequence of the araB gene from Lactobacillus plantarum is shown in SEQ ID NO: 2; the nucleotide sequence of the araD gene from Lactobacillus plantarum is shown in SEQ ID NO: 3.

[0061] The process of introdu...

Embodiment 1

[0095] Construction of integrated plasmids: araA (PA-araA) gene of Pediococcus acidilactici (also referred to as PA), araB (LP-araB) and araD (LP-araD) of Lactobacillus plantarum (also referred to as LP) ) gene was amplified, and the expression cassettes of three genes were respectively constructed: ENO2p-araD-SLM5t (nucleotide sequence as shown in SEQ ID NO: 4), GPM1p-araB-FBA1t (nucleotide sequence as shown in SEQ ID NO: 5), TEF1p-araA-CYC1t (the nucleotide sequence is shown in SEQ ID NO: 6); then the resistance gene expression cassette and three gene expression cassettes were assembled in series by the Golden Gate method, and multiple copies of the Saccharomyces cerevisiae genome were selected The sequence rDNA site is used as the integration site, and the constructed plasmid sequentially contains the sequence of the upstream 1000bp homology arm of rDNA, the bleomycin Zeocin resistance gene, the araA expression cassette, the araB expression cassette, the araD expression cass...

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Abstract

The invention relates to the fields of genetic engineering and metabolism engineering, discloses the fields of genetic engineering and metabolism engineering, and particularly relates to a reorganizedsaccharomyces cerevisia bacterial strain and a preparation method and application thereof. The reorganized saccharomyces cerevisia bacterial strain can express gene expression boxes of genes (1) to (3): (1) araA genes from pediococcus acidilactici; (2) araB genes from lactobacillus plantarum; and (3) araD genes from plant bacterium lacticum. The reorganized saccharomyces cerevisia bacterial strain has arabinose integrated exogenously as a metabolism way of a carbon source, in a technology for preparing biofluid fuel ethanol through fermentation of lignocellulose raw materials, glucose can beused, and arabinose can also be used, conversion rate of 93.2% of sugar alcohol in the lignocellulose raw materials can be realized, the bacterial strain is favorable in fermentability, the yield of ethanol in each ton of the raw materials is increased, the raw material utilization factor is increased, and the reorganized saccharomyces cerevisia bacterial strain is suitable for mass production ofcellulosic ethanol.

Description

technical field [0001] The invention relates to the fields of genetic engineering and metabolic engineering, in particular to a recombinant Saccharomyces cerevisiae strain and its preparation method and application. Background technique [0002] Fuel ethanol is recognized as the most promising new bioenergy. Because the production of the first-generation fuel ethanol has the problem of competing with others for food, a second-generation fuel ethanol production strategy using lignocellulosic materials as raw materials is proposed to promote the large-scale application of fuel ethanol. Since the 1970s, the utilization of total sugar in biomass has become an important topic in the development of renewable energy from lignocellulosic raw materials. Developing a production process to fully convert lignocellulosic raw materials into monosaccharide components and converting all obtained monosaccharide components into ethanol is a necessary condition for the production of ethanol b...

Claims

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Application Information

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IPC IPC(8): C12N15/81C12N1/19C12P7/10C12R1/865
CPCC12N15/81C12P7/10C07K14/195C07K14/335Y02E50/10
Inventor 佟毅李凡王康何太波张媛王小艳王燕李义袁敬伟刘辉
Owner JILIN COFCO BIOCHEM
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