Abstract: Twenty four strains for production of sterol esterase were screened from soil by using a plate assay in a medium containing sterol ester as a substrate and rhodamine B as the fluorescent. Among these strains, the jxpLT, which was found to produce the highest level of sterol esterase, belongs to genus Chryseobacterium based on 16S rDNA sequencing analysis and then was designated as Chryseobacterium sp. jxpLT. The maximum enzyme activity of this strain was 0.194 IU/mL after 18 h incubation under the optimized culture conditions. The sterol esterase was preliminary purified by ion exchange chromatography on DEAE Sepharose Fast Flow column and its enzymatic properties were investigated. This enzyme showed an optimum activity at pH 7.5 and 40℃. The enzyme had a good stability below 45℃ and the pH in 5.0-9.0. Chryseobacterium sp. jxpLT sterol esterase can hydrolyze cholesterol esters and PNP esters of different fatty acids. It may have the potential to solve the pitch problems caused by lipophilic extractives during wood mechanical pulping process.