Abstract: CFEM proteins, unique to fungi, play a crucial role in fungal growth, development, and pathogenicity. This study conducted a bioinformatics analysis based on CFEM proteins from 16 plant pathogenic fungi belonging to the Colletotrichum genus, aiming to clarify the physicochemical characteristics of these proteins. We searched the Pfam database using the keyword "CFEM" and obtained CFEM protein sequences from 16 Colletotrichum species. The presence of the CFEM domain was verified using SMART, and the sequences of CFEM proteins were finalized. Online programs such as protein databases and Expasy were utilized to analyze the physicochemical properties of these protein sequences. The results indicated that CFEM proteins from the Colletotrichum genus are primarily basic, stable, and hydrophobic proteins, with subcellular localization primarily at the plasma membrane. Their secondary structure is characterized by a relatively low number of beta sheets. The types of conserved domains include three categories: CFEM, CFEM+TM, and WSC+CFEM, all proteins possess conserved CFEM conserved domains, and the CFEM conserved domains sequences typically consist of 60-66 amino acids, including seven evenly spaced cysteine residues, exhibiting a high degree of conservation. This study provides insights into the function and mechanism of CFEM proteins in plant pathogenic fungi of the Colletotrichum genus.