Catalases are antioxidant enzymes which are responsible for decomposition of hydrogen peroxide to water and oxygen. Catalase activities have been shown to be influenced by environmental factors and stress conditions. In this study, in silico analysis of Nicotiana tabaccum L. was performed via bioinformatic tools. The results of this sudy suggested that the ORF of NtCAT-1 gene is 1479 bp and encodes a polypeptide of 492 amino acids. The predicted polypeptide was revealed as a 56.82 kDa protein with a pI of 6.27. The polypeptide had an aliphatic index of 71.52 and the grand average of hydropathicity (GRAVY) of -0.519. NtCAT-1 protein is hydrophilic and localised in Peroxisome. NtCAT-1 had two conserved domains at the positions of 18-399 and 421 and 486. had the catalase activity motif (CAM) at the position of 54–70 and heme-binding site (HBS) at the position of 344– 352. A highly reliable 3D structure was obtained and from Ramachandran plot analysis it was found that the portion of residues falling into the most favoured regions was 97.23%. The results of this study will provide fundamental information for further research in silico studies on catalase protein in different plant species.