Abstract: In this study, the glutathione peroxidase（GPX） of Lentinula edodes（LeGPX） was identified and cloned via coding sequence, and the protein bioinformation and gene expression pattern of LeGPX were analyzed. The results showed that a Le GPX gene was successfully identified and cloned, the open reading frame（ORF） of LeGPX gene was 621 bp, and the encoding protein contained 206 amino acid residues. The phylogenetic analysis showed that LeGPX protein was closely related to GPX in Marasmius fiardii and Moniliophthora roreri, and homology comparison showed that LeGPX protein contained characteristic sequence and catalytic site of GPX. Bioinformatic analysis showed that LeGPX was a hydrophilic unstable protein with molecular weight of 22.81 kD and theoretical isoelectric point of 8.83. The protein presented mainly random curly secondary structure without signal peptide and transmembrane structure. Le GPX soluble protein was obtained by prokaryotic expression and purification, and SDS-polyacrylamide gel electrophoresis（SDS-PAGE） showed that the protein size was in accordance with expectations. Fluorescence quantitative PCR analysis showed that the expression level of LeGPX gene was increased rapidly during L. edodes browning, and the LeGPX gene expression in mushroom stipe was more active compared with that in the cap.