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N-acetylneuraminic acid aldolase as well as coding gene and application thereof

A coding gene and coding technology, applied in application, genetic engineering, plant genetic improvement, etc., can solve the problems of small content and difficult separation of N-acetylneuraminic acid aldolase

Inactive Publication Date: 2013-04-24
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although N-acetylneuraminic acid aldolase exists in higher animals and some microorganisms, its content in organisms is very small, and it is difficult to separate and obtain a sufficient amount of N-acetylneuraminic acid aldolase

Method used

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  • N-acetylneuraminic acid aldolase as well as coding gene and application thereof
  • N-acetylneuraminic acid aldolase as well as coding gene and application thereof
  • N-acetylneuraminic acid aldolase as well as coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1, Preparation and functional verification of N-acetylneuraminic acid aldolase

[0032] 1. Extraction of Staphylococcus aureus genomic DNA and PCR amplification of SaNanA gene

[0033] Staphylococcus aureus (purchased from the Culture Collection Center of the Institute of Microbiology, Chinese Academy of Sciences, CGMCC 1.1529), and the bacterial genome extraction kit (Tiangen Company) was used to extract the genomic DNA of Staphylococcus aureus. Using the total genomic DNA extracted from Staphylococcus aureus as a template and Sa-F and Sa-R as primers, the SaNanA gene was amplified by PCR with high-fidelity pyrobest enzyme (Takara Company). The primer sequences are as follows:

[0034] Sa-F: 5'-GCGCGAC CATATG AACAAAGATTTAAAAGG-3' (the underline indicates the NdeI restriction site),

[0035] Sa-R: 5'-GCG GGATCC CTATAAATCGTATTTTGCAATG-3' (the underline indicates the BamHI restriction site).

[0036] The PCR program is:

[0037]

[0038] A fragment of a...

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Abstract

The invention discloses an N-acetylneuraminic acid aldolase as well as a coding gene and an application thereof. The N-acetylneuraminic acid aldolase provided by the invention is a protein of the following a) or b): a) a protein consisting of an ammonia acid sequence represented by a sequence 2 in a sequence table; and b) a protein with an activity of N-acetylneuraminic acid aldolase, wherein the ammonia acid sequence represented by the sequence 2 is subjected to substitution and / or deletion and / or addition of one or more of ammonia acid residues so as to obtain the protein derived by the protein in the a). According to the invention, a new gene SananA for coding the N-acetylneuraminic acid aldolase is obtained, and the gene SananA is cloned to a prokaryotic expression vector 6HisT-pRSET plasmid, so that a high-efficiency expression of a N-acetylneuraminic acid aldolase protein (SaNanA) can be carried out in escherichia coli. The obtained recombinant N-acetylneuraminic acid aldolase protein provided by the invention can catalyze N-acetylmannosamine and pyruvic acid to generate N-acetylneuraminic acid, so that the N-acetylneuraminic acid aldolase protein has high enzyme activity, application foreground and development value.

Description

technical field [0001] The invention relates to N-acetylneuraminic acid aldolase, its coding gene and application. Background technique [0002] Sialic acid (Sialic acid) is a kind of neuraminic acid derivatives, and more than 40 natural sialic acid derivatives have been found so far. The most important of these is N-acetylneuraminic acid (N-acetyl-D-neuraminic acid, Neu5Ac). Neu5Ac is usually located at the non-reducing end of the sugar chain of glycoproteins and glycolipids on the cell surface, and plays an important role in many physiological processes related to the interaction of sugars and proteins: such as cell adhesion, signal transmission, and stability of glycoproteins sex, bacterial pathogenicity, viral infection, inflammation and tumor metastasis, etc. The biological functions of N-acetylneuraminic acid are diversified, and it has important medical value in the treatment of influenza, neurological diseases, inflammation and tumors, especially in the prevention ...

Claims

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Application Information

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IPC IPC(8): C12N9/88C12N15/60C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21C12N15/11C12P19/26
Inventor 林白雪张子娟陶勇
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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