An efficient di-n-butyl phthalate（DBP） degrading bacterial strain Aci-17 was isolated from the fermentation grain of Baijiu. Based on its 16 S rRNA gene sequence， strain Aci-17 was identified as Acinetobacter indicus sp. The ester bond hydrolase gene in strain Aci-17 was cloned by gene recombination， aimed to gain insight into the mechanism of phthalate ester （PAEs） degradation. The recombinant Est96 protein demonstrated that it exhibits excellent capacity in degrading for PAEs， the degradation rate of dimethyl phthalate（DMP） reached 99.996%. The bioinformatics analysis of sequencing result was systematically performed， the Est96 encoded by gene Est96 which has the typical conserved motif G-X-S-X-G and HGGG， they are belonged to family IV of hydrolases. the Est96 exhibited a predicted molecular weight of 40.66 ku， amino acid residues 201 is the active site. Docking results of Est96 with DMP show the main interaction forces are the hydrogen bond， van der waals force and Pi-Pi interaction. The conserved sequence motif HGGG， GDSAG and HGF affect the hydrolysis of the substrate ester bond， and laid the foundation for the further exploring the mechanism of Acinetobacter esterase-catalyzed biodegradation of PAEs.