Hypothetical proteins are predicted or experimentally uncharacterized proteins and they constitute a substantial portion of the proteome of both eukaryotes and prokaryotes . With the remarkable advancement in the field of Next Generation Sequencing (NGS), the number of hypothetical proteins is increasing rapidly and comparing to that experimental validation rate is not so high. This gap of structural and functional annotation can be reduced through in silico approach using modern bioinformatic tools which might pave the way for new drug synthesis and vaccine development. Thus, the current study focuses on annotating AVO28_00330 hypothetical protein of Y. pestis, both structurally and functionally for an improved understanding, which might help later at drug and vaccine development.