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Yeast strain for degrading long-chain alkane and application thereof

A technology of alkanes and bacterial strains, applied in the field of microorganisms, can solve the problems of low degradation efficiency of long-chain alkanes and difficulty in meeting the requirements of alkane degradation, and achieve the effect of improving the efficiency of alkane degradation

Pending Publication Date: 2022-07-29
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the alkane-degrading engineering bacteria based on Saccharomyces cerevisiae have low efficiency in degrading long-chain alkanes, and it is difficult to meet the industrial requirements for alkane degradation.

Method used

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  • Yeast strain for degrading long-chain alkane and application thereof
  • Yeast strain for degrading long-chain alkane and application thereof
  • Yeast strain for degrading long-chain alkane and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0083] Example 1 Construction of a lipolytic chassis for strengthening the alkane degradation pathway

[0084] The present invention selects Yarrowia lipolytica, and exogenously expresses AlmA1 and AlkM derived from Acinetobacter sp. ADP1, LadA alkane hydroxylase gene derived from Geobacillus thermodenitrificans NG80-2, and AlmA2 derived from Acinetobacter sp. M-1, respectively. To achieve efficient degradation of long-chain alkanes. Degradation of long-chain alkanes by Yarrowia lipolytica figure 1 shown.

[0085] 1. Screening of alkane hydroxylases

[0086] In this study, Yarrowia lipolytica ATCC 201249 was selected as the starting strain, and the codon-optimized alkane hydroxylase (AlmA1, AlkM, LadA and AlmA2) genes of Yarrowia lipolytica were integrated into the PUC57 vector. BsaI restriction site. After digestion with BsaI, the digestion system was subjected to gel electrophoresis, and agarose gel recovery kit was used for fragment recovery to obtain linear fragments A...

Embodiment 2

[0101] Example 2 Construction of a lipolytic chassis for strengthening the alkane transport pathway

[0102] 1. Overexpression of alkane transporters

[0103] The current research on the mechanism of alkane degradation is not only the first step of the hydroxylation process of alkanes entering cells, but also focuses on the study of genes encoding alkane transporters. Protein-mediated transport of small molecules across the cytoplasmic membrane is important for the import and export of hydrophobic or lipid compounds. Although membrane transporters are involved in one-third of remodeled metabolic networks in yeast, they are still widely ignored. In Yarrowia lipolytica, the alkane transporter ABC1, similar to the yeast pleiotropic drug resistance transporter PDR5, was identified as required for alkane uptake.

[0104] Therefore, we overexpressed lipolytic yeast endogenous alkane transporter (ABC1) in wild-type lipolytic yeast and engineered yeast YAH06, hoping to further impro...

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Abstract

The invention relates to the technical field of microorganisms, in particular to a yeast strain for degrading long-chain alkane and application of the yeast strain. The Yarrowia lipolytica is used as a chassis, an exogenous alkane hydroxylase gene module is integrated with a yeast genome, and the alkane hydroxylase exogenous single-copy expression yeast strain is obtained. On the basis, the strain is optimized, the degradation efficiency of the obtained alkane hydroxylase exogenous multi-copy expression yeast strain can reach 71.50%, the activity of the alkane degrading enzyme is 0.65 U / mg, and the degradation efficiency is relatively high when glucose is used as a substrate; the alkane degradation efficiency of the obtained strain formed by combining the endogenous alkane transporter and the exogenous alkane hydroxylase gene module is further improved, the degradation efficiency can reach 75.12%, and the hydrophobicity is 91.8%. In addition, the alkane degradation efficiency of the exogenous gene module integrated strain is superior to that of the endogenous gene module integrated strain.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a yeast strain for degrading long-chain alkanes and its application. Background technique [0002] With the rapid and vigorous development of my country's economy and society, the demand and consumption of energy are also increasing. However, the incidence of various oil spills is also increasing, causing immeasurable damage to our country's environment and economy. With the enhancement of environmental awareness, the research on bioremediation of petroleum hydrocarbon environmental pollution has been continuously strengthened. Compared with the commonly used physical and chemical remediation methods, bioremediation has received more and more attention due to its advantages such as high efficiency, environmentally friendly by-products, and low cost. In recent years, genetically engineered bacteria transformed by systems biology strategies have achieved biodegradation of ...

Claims

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Application Information

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IPC IPC(8): C12N15/53C12N15/31C12N15/81C12N1/19C12R1/645A62D101/20
CPCC12N9/0004C07K14/39C12N15/815A62D3/02C12R2001/645A62D2101/20Y02A20/204
Inventor 元英进王妍丁明珠曹志北
Owner TIANJIN UNIV
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