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Modified akkermansia muciniphila Amuc_1100 protein and preparation method and application thereof

A protein and amino acid technology, applied in the field of protein engineering, can solve the problems of high-efficiency protein drugs, unclear molecular mechanism, inability to develop and other problems, and achieve the effect of improving biological activity and good application prospects.

Active Publication Date: 2020-04-03
ANHUI UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • Modified akkermansia muciniphila Amuc_1100 protein and preparation method and application thereof
  • Modified akkermansia muciniphila Amuc_1100 protein and preparation method and application thereof
  • Modified akkermansia muciniphila Amuc_1100 protein and preparation method and application thereof

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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 present invention discloses a modified akkermansia muciniphila Amuc_1100 protein and a preparation method and an application thereof, and belongs to the technical field of protein engineering. Bymodifying the existing akkermansia muciniphila Amuc_1100 protein, 60-90 amino acids at a N-terminus of the Amuc_1100 protein is truncated all or partially, or mutation of some amino acids within the range of 60-90 amino acids at the N-terminus of the Amuc_1100 protein is conducted, both the truncated modification and the amino acid mutation can change aggregation state of the Amuc_1100 protein andalso improve biological activity of the Amuc_1100 protein, which provides a very good application prospect for 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 Applications(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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