Liesong Chen ^1,2 Zhuojia Zhang ^1,2* Qilin Zeng ^1,2 Wei Wang ^3* Hui Zhou ^4* Yimou Wu ^1*

• ¹ Institute of pathogenic biology, Hengyang, China
• ² Hunan Provincial Key Laboratory for Special Pathogens Prevention and Control, University of South China, Hengyang, Hunan Province, China
• ³ Special Inspection Department, Hengyang traditional Chinese Medicine Hospital, Hengyang, Hunan, CHina, Hengyang, Hunan Province, China
• ⁴ The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan, China

Ureaplasma urealyticum, which is commonly present in the human lower genitourinary tract as a commensal organism, has been associated with various urogenital infections along with several complications in certain individuals. Increasing issue of antibiotic resistance, coupled with the scarcity of vaccines, underscores the necessity to explore novel drug targets for effectively treating U. urealyticum infection. In this research, we focus on identifying novel drug target proteins against U. urealyticum using a subtractive genomics approach integrated with comparative metabolic pathway analysis. Through diverse subtractive genomics methods, we systematically narrowed down the complete proteomes of thirteen shortlisted Ureaplasma strains to two target proteins, which we proposed as novel therapeutic targets. Subsequently, these two target proteins, designated B5ZC96 and B5ZAH8, were selected for further structure-based studies. These proteins could serve as a starting point for developing lead drug candidates that could potentially inhibit them and reduce the risk of drug-resistant U. urealyticum infection. Ultimately, targeting these enzymatic proteins could pave the way for targeted therapy against U. urealyticum.