In this paper, we report Bacillus subtilis TLO3, which was isolated from olive tree rhizosphere and exhibits high amylolytic activity. The genome of Bacillus subtilis TLO3 contains 4,071 protein-coding sequences carried on one chromosome (4,232,155 bp) with an average G+C content of 44.1% and 119 RNA molecules. The gene encoding α-amylase was detected, as well as other genes related to starch and cellulose hydrolysis, making the strain a potential candidate for industrial treatment of starch and lignocellulose biomasses.