The eukaryotes sterol pathways are extremely conserved and these biosynthetic pathway are very long which includes the synthesis of dolichols, coenzyme Q, heme A, and isoprenylated proteins.14-Demethylase is an essential enzyme of the cytochrome P450 superfamily, which is potential the target of azole antifungals. Predicted results shows that 14-alpha sterol demethylase have molecular weight of 58930.8 Daltons and the theoretical isoelectric point (pI) of 7.64. The negative Grand average of hydropathicity (GRAVY) index of ‐0.125. The Aliphatic index of Aspergillus fumigates 14-alpha sterol demethylase is 89.48. Alpha helix (Hh) accounts 210 amino acids of about 40.08%. The extended strand (Ee) had 91 amino acids accounting 17.37, Beta turn (Tt) made up of 51 amino acids making up 9.73% and random coil (Cc) made up of 172 amino acids accounting 32.82%. The subcellular localization of 14alpha sterol demethylase Cyp51B was predicted to be a Plasma membrane protein.