Method for expressing trehalose synthase by Corynebacterium glutamicum through an EF-Tu promoter and application of method

A technology of Corynebacterium glutamicum and trehalose synthase, applied in the field of bioengineering, can solve the problems of increasing production cost, increasing technological process and control difficulty, etc.

Inactive Publication Date: 2021-07-23
山东恒仁工贸有限公司
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Problems solved by technology

[0004] In the research of enzymatic transformation and synthesis of trehalose in the prior art, trehalose synthase is expressed by recombining the plasmid into Escherichia coli, but when the recombinant strain is used to express the protein, the promoters of the commonly used plasmids for protein expression are inducible

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  • Method for expressing trehalose synthase by Corynebacterium glutamicum through an EF-Tu promoter and application of method
  • Method for expressing trehalose synthase by Corynebacterium glutamicum through an EF-Tu promoter and application of method
  • Method for expressing trehalose synthase by Corynebacterium glutamicum through an EF-Tu promoter and application of method

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[0023] In order to make the object, technical solution and advantages of the present invention more clear, the present invention will be further described in detail below in conjunction with the examples. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0024] 1. Material selection

[0025] The used Corynebacterium glutamicum model strain ATCC13032 was purchased. C. glutamicum ATCC13032P EF-Tu The promoter sequence is shown in SEQ ID NO.1, and the Cgtrs sequence of the C. glutamicum ATCC13032 trehalose synthase gene is shown in SEQ ID NO.2.

[0026] 2. Primer Design

[0027] Using PCR amplification technology, the C. glutamicum ATCC13032 genome was used as a template to amplify P EF-Tu Promoter gene fragment and trehalose synthase gene Cgtrs. The sequence listing 1 of the design is as follows:

[0028] Table 1: Primer sequences

[0029]

[0030] 3. Gene recombina...

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Abstract

The invention discloses a method for expressing trehalose synthase by Corynebacterium glutamicum through an EF-Tu promoter and application of the method. The method comprises the following steps: fusing the promoter PEF-Tu of a translation elongation factor EF-Tu in Corynebacterium glutamicum ATCC13032 to the upstream of a trehalose synthase gene of Corynebacterium glutamicum ATCC13032, and cloning to a Corynebacterium glutamicum induced expression plasmid pXMJ19, and the trehalose synthase can be over-expressed in the Corynebacterium glutamicum without induction and used for converting maltose to synthesize trehalose. According to the invention, the corynebacterium glutamicum endogenous strong constitutive expression promoter is utilized to express the target protein, overexpression can be realized without induction, and the production cost can be reduced in actual production.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a method and application for expressing trehalose synthase by Corynebacterium glutamicum using EF-Tu promoter. Background technique [0002] Trehalose is a non-reducing disaccharide composed of α,α-1,1-glucosidic bonds linking glucose molecules, and its stability is far superior to other small sugar molecules such as maltose, sucrose, and glucose. Trehalose has a specific protective effect on biological tissues, cells and biological macromolecules. It can be used to maintain cell activity, prevent food deterioration, maintain food freshness, improve food quality, and enhance the function of moisturizing cosmetics. Therefore, trehalose has broad application prospects in the pharmaceutical, food and cosmetic industries. [0003] Trehalose can be produced by chemical synthesis, microbial fermentation and enzymatic conversion. The production of trehalose by chem...

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Application Information

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IPC IPC(8): C12N15/77C12N15/67C12N15/61C12N1/21C12R1/15
CPCC12N15/77C12N9/90C12Y504/99016
Inventor 满在伟崔慧慧刘典典陈一鸣蒋心怡郭静
Owner 山东恒仁工贸有限公司
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