A modified Ekkermansia amuc_1100 protein and its preparation method and application

A protein, A85E technology, applied in the field of protein engineering, can solve problems such as unclear molecular mechanism, high-efficiency protein drugs, and failure to develop them

Active Publication Date: 2021-05-04
ANHUI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Although the current research results show that Amuc_1100 protein can inhibit obesity, type 2 diabetes and improve human intestinal immune function, but its molecular mechanism is not clear, so it is impossible to develop more efficient protein drugs

Method used

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  • A modified Ekkermansia amuc_1100 protein and its preparation method and application
  • A modified Ekkermansia amuc_1100 protein and its preparation method and application
  • A modified Ekkermansia amuc_1100 protein and its preparation method and application

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Embodiment 1

[0031] 1. Materials

[0032] The methods used in this example are conventional methods known to those skilled in the art unless otherwise specified, and the reagents and other materials used are all commercially available products unless otherwise specified.

[0033] 2. Method

[0034] 2.1 Construction, expression and purification of Ekkermansia Amuc_1100 protein (Amuc_1100)

[0035] The present invention uses gene synthesis technology to obtain the gene of Amuc_1100 protein, constructs the gene on the pET-22b vector, utilizes the Escherichia coli prokaryotic expression system to express Amuc_1100 protein in large quantities, and uses Ni-NTA column purification to obtain high-purity recombinant protein, specifically Steps include:

[0036] 2.1.1 Construction of recombinant plasmids

[0037] (1) According to the nucleotide sequence of the wild-type Amuc_1100 protein coding gene shown in SEQ ID NO.2, the Amuc_1100 protein gene was synthesized using gene synthesis technology, ...

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Abstract

The invention discloses a modified Ekkermansia Amuc_1100 protein and its preparation method and application, belonging to the technical field of protein engineering, through the modification of the existing Ekkermansia Amuc_1100 protein, including all or part of the Amuc_1100 protein N-terminal truncation The transformation of the first 60‑90 amino acids, or the mutation of some amino acids within the range of 60‑90 amino acids at the N-terminal of the Amuc_1100 protein, found that whether it is truncation modification or amino acid mutation, it can change the aggregation state of the Amuc_1100 protein, and at the same time improve The biological activity of Amuc_1100 protein has been confirmed, which provides a good application prospect for the development of more efficient drugs.

Description

technical field [0001] The present invention relates to the technical field of protein engineering, in particular to a modified Ekkermansia Amuc_1100 protein and its preparation method and application. Background technique [0002] Akkermansia muciniphila (A. muciniphilas) is a mucin-degrading gut microorganism discovered in 2004 that colonizes the mucus layer of the human intestine1. Recent research results have shown that A. muciniphilas can alleviate the symptoms of obesity and metabolic diseases2-4, making it gradually become a new type of beneficial bacteria for human health research. While the research on A. muciniphilas is gradually deepening, its outer membrane protein is gradually being paid attention to. Amuc_1100 protein is an outer membrane protein of A.muciniphilas discovered in 20165. Research results show that this protein has the same effect as A.muciniphilas on reducing mouse body weight and improving mouse glucose and insulin tolerance6, In addition, Amuc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/195C12N15/70A61K38/17A61P3/04A61P3/10A61P1/00
CPCA61K38/00A61P1/00A61P3/04A61P3/10C07K14/195C12N15/70
Inventor 张敏王明珠王俊超相蕊
Owner ANHUI UNIVERSITY
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