Abstract: The present disclosure relates to a composition and a kit for colonization of external probiotics in the gut through bioorthogonal click chemistry. The composition or kit for colonization of external probiotics in the gut according to the present disclosure has the effects of overcoming colonization resistance, which is the limitation of existing external probiotics caused by physical discharge and indigenous bacteria, by linking surface-modified external probiotics to the intestinal mucosa into which a bioorthogonal functional group has been introduced, and beneficially changing the intestinal environment by long-term colonization of various external probiotics, such as lactic acid bacteria, bifidobacteria, and yeast. Accordingly, the present disclosure is expected to be effectively used for the development of various medicines and functional foods using probiotics.