Method for carrying out bio-orthogonal polysaccharide labeling from microbiome level, and applications thereof

Abstract

The present invention provides a method for carrying out bio-orthogonal polysaccharide labeling from a microbiome level. The method comprises: placing microbial flora in a culture medium added with afirst probe, and culturing to obtain a microbial flora labeled with the first probe, wherein the first probe is one or a variety of unnatural saccharides; and carrying out a bio-orthogonal reaction onthe microbial flora labeled with the first probe and a second probe to obtain a microbial flora labeled with the second probe, wherein the second probe is a chemical marker capable of being subjectedto a bio-orthogonal reaction with the first probe. According to the present invention, with the method, a variety of bacteria capable of being metabolized and labeled with unnatural saccharides can be rapidly screened in the high-throughput manner, the specific types of the unnatural saccharides for metabolizing and labeling different bacteria can be determined, the possibility can be provided for the in vivo imaging of the specific microbiome, and the powerful tool is provided for the exploring of the biological functions of the macromolecules containing the polysaccharide structure in the bacteria difficultly cultured separately.

Description

technical field [0001] The invention relates to the field of chemical biology, in particular to a method and application of bio-orthogonal glycan labeling at the microbiome level. Background technique [0002] Using chemical biology methods as an entry point, labeling cellular glycan structures by introducing precursor monosaccharide analogs (non-natural sugars) containing bioorthogonal groups such as alkyne or azide during the biosynthesis of glycans the goal of. Through the design of non-natural sugar probes, they can enter the sugar biosynthesis mechanism of cells, pass through cell metabolism, and finally be expressed on the glycan-containing structures on the cell surface. Finally, probe molecules, such as fluorescent probes and biotin probes, can be introduced under living cell conditions through bioorthogonal reaction. Bioorthogonal glycan labeling has also been extensively reported in bacteria. In several representative studies, different non-natural sugar probes c...

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Problems solved by technology

[0005] However, the reports on bio-orthogonal glycan labeling in bacteria are limited to screening for a specific bacteria that can be cultured alone, the efficiency is low, and for bacteria that cannot be cultured alone (generally considered to account for more than 95% of all bacteria) ) helpless

The same problem exists in the discovery of new glycoproteins in bacteria. It is often impossible to study bacteria that cannot be cultured alone, and it is impossible to understand the potential glycoproteins.

[0006] The above-mentioned commonly used methods for microbiome imaging have the following problems (1) The disadvantage of the method of inserting fluorescent proteins is that >80% of intestinal commensal microorganisms cannot be cultured in vitro alone, and thus cannot be transgenic

(2) FISH has problems such as complex DNA probe design, low rRNA content, cumbersome operation, etc., and is limited to the detection of bacteria whose existence is known in advance and whose primer feasibility has been verified, so its application is still subject to many restrictions

(3) The method of using chemical biology to label intestinal commensal bacteria has the disadvantage that specific bacteria still need to be cultured separately

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