The honey bee species Apis mellifera, commonly known as Italian honeybee, is a prominent pollinator among other species. These are found to be associated with the symbionts in their gut, which make them a powerful organism. The present investigation on the culturing of the Apis mellifera gut bacteria and their isolation for the identification of the phylum of the bacteria. It was observed that three media type namely, Nutrient Agar, Tryptone soy Agar and Actinomycetes isolation agar were most suitable one by using spread plate method. The pure bacterial culture plates were obtained streaking the individual colony. Form the individual colony 16S rRNA gene sequence analysis showed that A. mellifera gut is a home for Firmicutes, Proteobacteria and Actinobacteria.