The present study is aimed to synthesize V2O5 using V(ORi)3 precursor and study its anti-microbial activity. The V(ORi)3 was prepared by reaction of VCl3 and isopropanol alcohol. The synthesized V(ORi)3 was hydrolyzed to yield black V(OH)3 gel using concentrated HCl as gelling agent. Yellow V2O5 obtained by thermal treatment of V(OH)3 gel at110oC and 380oC. All products were characterized using FT-IR Spectroscopy method. IR spectral revealed to found V(ORi)3, V(OH)3 gel and V2O5 were formed. The V2O5 has been tested in vitro against numbers of microorganisms, to assess their antimicrobial activity using disc diffusion method, according to the National Committee for Clinical Laboratory Standards Guidelines. The results showed that V2O5 is active against Staphylococcus aureus and Pseudomonas aeruginosa and partial active against Bacillus subtilis, Escherichia coli and Candida albicans. Antimicrobial Study of VO(OR)3 is required.