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A method for enzyme-catalyzed selective amidation and hydrazide modification of polypeptide C-terminus

A C-terminal and hydrazide technology, which is applied in the field of enzyme-catalyzed selective amidation and hydrazide modification of polypeptide C-terminus, can solve the problems of low yield, high difficulty, and many chemical modification steps, and achieve substrate spectrum Broad, mild effect

Active Publication Date: 2022-05-06
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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AI Technical Summary

Problems solved by technology

[0003] Due to the complexity of the polypeptide structure, there are many chemical modification steps, low yield and great difficulty

Method used

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  • A method for enzyme-catalyzed selective amidation and hydrazide modification of polypeptide C-terminus
  • A method for enzyme-catalyzed selective amidation and hydrazide modification of polypeptide C-terminus
  • A method for enzyme-catalyzed selective amidation and hydrazide modification of polypeptide C-terminus

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Embodiment 1

[0034] Example 1. Enzyme-catalyzed selective amidation and hydrazide modification of polypeptide C-terminus

[0035] In this example, the amino acid at position 1 of the C-terminus of the polypeptide is selectively amidated and hydrazidated using the polypeptide terminal amidation enzyme (PAM12B) shown in SEQ ID No.1.

[0036] Reaction process: The C-terminally amidated polypeptide to be modified is catalyzed by PAM12B shown in SEQ ID No.1 and nucleophilic organic amine (NH 2 R) reaction, thereby realizing the amidation and hydrazide modification of the C-terminal amino acid of the polypeptide to be modified.

[0037] Wherein, the general formula of the nucleophilic organic amine with the modifying group is NH 2 R can specifically be any of the following:

[0038]

[0039] 1. Preparation of polypeptide terminal amidation enzyme (PAM12B)

[0040] Adopt the double-stranded DNA molecule shown in SEQ ID No.2 (the coded gene of polypeptide end amidating enzyme PAM12B shown in...

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Abstract

The invention discloses an enzyme-catalyzed C-terminal selective amidation and hydrazide modification method of polypeptide. The present invention provides a method for amidation and / or hydrazide modification of the C-terminal amino acid of a polypeptide, which is to use a specific protein to perform amidation and / or hydrazide modification of the C-terminal amino acid of the polypeptide; the specific protein is SEQ The protein indicated by ID No.1. Using this method, the amidation and / or hydrazide modification of the C-terminal amino acid of the polypeptide can be realized, and it is an aqueous phase reaction with mild conditions, and the C-terminus is selectively modified without affecting other modification groups on the side chain, and the side chain does not need to be protected , wide substrate spectrum, no restrictions on the amino acid sequence of the substrate polypeptide, any amino acid except proline at the end of the modified polypeptide can be modified.

Description

technical field [0001] The invention relates to the field of polypeptide chemistry, in particular to an enzyme-catalyzed C-terminal selective amidation and hydrazide modification method of polypeptide. Background technique [0002] The terminal functionalization of a polypeptide has a very important impact on its biochemical properties. Through the specific modification of the C-terminus, the in vivo metabolic half-life of the polypeptide can be prolonged, and the immunogenicity or toxic side effects can be reduced. In the study of the biochemical properties of peptides, researchers often need to add different functional groups (for example, fluorescent groups, synthetic polymers or immunolabels) to the peptide structure. In addition, efficient and accurate peptide C-terminal protection, deprotection and selective activation methods are also urgently needed technologies in industrial peptide synthesis and production. Therefore, the selective C-terminal modification method ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/00
CPCC12P21/00
Inventor 吴边李瑞峰田玉娥
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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