A recombinant microorganism, a preparation method thereof and its application in the production of coenzyme Q10. Specifically, the present disclosure provides a method of exogenously introducing a gene encoding the global regulatory protein irrE to construct a recombinant microorganism. This recombinant microorganism is suitable for producing coenzyme Q10 by fermentation method, and is particularly suitable for producing oxidized coenzyme Q10. The recombinant microorganism of the present disclosure has stress resistance, and has good tolerance against harsh environments including high osmotic pressure and high redox potential, thus making it possible to significantly increase the yield of coenzyme Q10, especially the yield of oxidized coenzyme Q10.