Abstract: To explore the physicochemical properties of pathogenic gene protein sequences from different categories of Colletotrichum gloeosporioides, we conducted a bioinformatics analysis was conducted based on 59 previously identified pathogenic gene protein sequences. The physicochemical properties of these proteins were analyzed using various online tools, including ProtParam, Protscale, ProtComp, Phyre, DeepTMHMM, SMART, and STRING. The 59 pathogenic genes were classified into proteins, enzymes, and regulatory factors categories for statistical summary. The results revealed that the theoretical isoelectric points were concentrated between 5.01 to 7.00, and the instability coefficients were mainly in the range of 30.01 to 50.00. Subcellular localization was predominantly in the nucleus, cytoplasm, and plasma membrane. Among the amino acid residues, K had the highest number of hydrophilic residues, while A had the highest number of hydrophobic residues. However, all enzymes and regulatory factors were hydrophilic proteins, with some proteins being hydrophobic. In terms of secondary structure, enzymes and proteins had a higher proportion of α-helices, while regulatory factors had a higher proportion of random coils. Regulatory factors also had a higher proportion of conserved domains, and protein interactions were mainly observed among enzymatic proteins.