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Engineered yeast and construction method and application thereof

A yeast engineering and yeast technology, applied in the fields of biotechnology and bioenergy, can solve the problems of low tolerance and inability of lignin degradation products, and achieve the effects of broad industrial application prospects, improved yield and efficient transformation

Active Publication Date: 2021-11-09
DALIAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Natural oleaginous yeast cannot biotransform lignin degradation products and have low tolerance to lignin degradation products
At present, there is no report on the use of metabolic engineering to achieve the biotransformation of lignin degradation products by oleaginous yeast to improve the tolerance of oleaginous yeast to lignin degradation products

Method used

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  • Engineered yeast and construction method and application thereof
  • Engineered yeast and construction method and application thereof
  • Engineered yeast and construction method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1 Construction, screening and application of Rhodosporidium toruloides haploid engineering strain expressing lignin multifunctional peroxidase VP and manganese peroxidase MNP

[0064] 1. Construction of Rhodosporidium toruloides haploid engineering strain expressing lignin multifunctional peroxidase VP

[0065] (1) Synthesis of lignin multifunctional peroxidase gene VP

[0066] The source of the VP gene is Pleurotus eryngii (GenBank ID: AF175710, the amino acid sequence of the encoded protein is shown in SEQ ID NO.1), and it is optimized according to the codon preference of Rhodosporidium toruloides haploid (the optimized VP gene sequence shown in SEQ ID NO.3), and the whole sequence was synthesized, and the synthesized gene was connected to the pUC57 vector to obtain the plasmid pUC57-VP.

[0067] (2) Construction of pZPK-VP vector

[0068] The target vector was constructed by RF cloning method. Plasmid pZPK-P PGK -HYG-P2A-BLE-T HSP (JiaoXiang, Zhang Qi, Z...

Embodiment 2

[0101] Example 2 Construction, Screening and Application of Hyphoderma Dermatoides Engineering Strain Expressing Lignin Multifunctional Peroxidase VP and Manganese Peroxidase MNP

[0102] 1. Construction and screening of engineering strains of Trichosporium dermatitis expressing lignin multifunctional peroxidase VP and manganese peroxidase MNP

[0103] Trichosporon cutaneum (Trichosporon cutaneum) CGMCC No.2.571 was used as the starting strain to construct engineering strains of Trichosporon cutaneum overexpressing MNP and VP genes respectively. The construction method was as follows: the recombinant plasmids pZPK-MNP and pZPK-VP were transferred into The engineering strain obtained from the starting strain, wherein, the construction process of pZPK-MNP and pZPK-VP expression vectors, the ATMT transformation process is as described in Example 1, and the electric shock competent preparation and transformation steps are as follows:

[0104] (1) Pick a single colony of the yeast ...

Embodiment 3

[0118] Example 3 Construction, screening and application of L. stariae engineering strains expressing lignin multifunctional peroxidase VP and manganese peroxidase MNP

[0119] 1. Construction and screening of engineering strains expressing lignin multifunctional peroxidase VP and manganese peroxidase MNP Staria oleoyeast

[0120] Using Lipomyces starkeyi NRRL Y-11557 as the starting strain, the recombinant plasmids pZPK-MNP and pZPK-VP were transformed into L. starkeyi to obtain engineering strains of Lipomyces starkeyi overexpressing MNP and VP, respectively. The construction process of pZPK-MNP and pZPK-VP expression vectors, the ATMT transformation process are as described in Example 1, the electroporation transformation process is as described in Example 2, the engineering strain screening steps are as described in Example 1, and the obtained engineering strain is named LS -MNP and LS-VP.

[0121] 2. Fermentation of the engineered strains LS-MNP and LS-VP of Lepidoptera ...

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Abstract

The invention relates to the technical field of biotechnology and biological energy, in particular to an engineered yeast and a construction method and application thereof. The engineered yeast expresses lignin multifunctional peroxidase and / or manganese peroxidase. The engineered yeast can grow, synthesize and accumulate microbial oil by using a lignin degradation product as a carbon source, can realize efficient conversion from a lignin biomass raw material to oil, provides a core technology for full-resource utilization and efficient conversion of the lignocellulose biomass raw material, and has a wide industrial application prospect.

Description

technical field [0001] The invention relates to the technical fields of biotechnology and bioenergy, in particular to a yeast engineering bacterium and its construction method and application. Background technique [0002] Oil is an important bulk product indispensable to human life, and it can replace fossil resources as the basic processing raw material for the chemical industry and renewable energy industry. In recent years, the global oil demand has been increasing day by day, resulting in shortage of oil resources and large price fluctuations. Microbial oil is an intracellular product synthesized by oil-producing microorganisms under specific conditions. It has attracted much attention because its production is not restricted by climate, season and site. Lipid biorefinery research using cheap and abundant lignocellulosic biomass resources as raw materials has attracted great attention in recent years, and is considered to be a new approach for oil production with poten...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/19C12N15/53C12N15/81C12P7/64C12R1/645
CPCC12Y111/01013C12Y111/01014C12N9/0065C12N15/815C12P7/6463C12P2203/00
Inventor 薛闯黄其田吕力婷程驰吴又多
Owner DALIAN UNIV OF TECH