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Recombinant Saccharomyces cerevisiae strain and its preparation method and application

A technology of yeast strain and Saccharomyces cerevisiae, which is applied in the field of genetic engineering and metabolic engineering, can solve the problems that the utilization rate of arabinose needs to be improved, and achieve the effects of improving raw material utilization rate, high sugar alcohol conversion rate, and increasing ethanol yield

Active Publication Date: 2022-02-18
JILIN COFCO BIOCHEM +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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

Method used

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  • Recombinant Saccharomyces cerevisiae strain and its preparation method and application
  • Recombinant Saccharomyces cerevisiae strain and its preparation method and application
  • Recombinant Saccharomyces cerevisiae strain and its preparation method and application

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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 with the deposit number CGMCC No.16830.

[0056] The third aspect of the present invention provides a method for preparing a recombinant yeast strain, comprising: introducing a 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 as 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 introducing the gene expression...

Embodiment 1

[0095] Construction of integrated plasmid: 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 constructed respectively: ENO2p-araD-SLM5t (nucleotide sequence as shown in SEQ ID NO: 4), GPM1p-araB-FBA1t (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 was used as the integration site, and the construction plasmid sequentially contained the sequence of the 1000bp homology arm upstream of the rDNA, the bleomycin Zeocin resistance gene, the araA exp...

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Abstract

The invention relates to the field of genetic engineering and metabolic engineering, discloses the field of genetic engineering and metabolic engineering, and specifically relates to a recombinant Saccharomyces cerevisiae strain and its preparation method and application. The recombinant Saccharomyces cerevisiae strain of the present invention comprises a gene expression cassette capable of expressing the genes of the following (1) to (3): (1) the araA gene from Pediococcus acidilactici, (2) the araA gene from Lactobacillus plantarum plantarum) araB gene, (3) araD gene from Lactobacillus plantarum. The recombinant Saccharomyces cerevisiae strain of the present invention has a metabolic pathway of exogenously integrated arabinose as a carbon source, and can utilize both glucose and arabinose in the process of preparing bioliquid fuel ethanol by fermenting lignocellulosic raw materials, and can realize lignocellulosic The sugar-alcohol conversion rate of 93.2% of the vegetarian raw material, and the fermentation performance of the bacterial strain are good, the ethanol production rate per ton of raw material is improved, the raw material utilization rate is improved, and the method is suitable for large-scale production of cellulosic 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 a preparation method and application thereof. Background technique [0002] Fuel ethanol is recognized as the most promising new bioenergy. Since the production of the first generation of fuel ethanol has the problem of competing with people for food, a strategy for the production of the second generation of fuel ethanol using lignocellulosic materials as raw materials is proposed to promote the large-scale application of fuel ethanol. Since the 1970s, the utilization of whole sugars from biomass has become an important issue 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 the obtained monosaccharide components into ethanol is a necessary condition...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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