The soil microbes were preliminarily screened by a simple and modified rapid screening method. The strains with significant change in color ring were selected to conduct a secondary screening through UV-visible spectrograph and one strain z123 capable of producing pancreatic lipase inhibitor was finally obtained. It was identified as Bacillus subtilis based on morphology, physiological and biochemical characterization, 16S rDNA sequence homology analysis and phylogenetic tree. The inhibition rate of the fermentation product reached 38.4%, which had great value for further study.