Purification and Biochemical Characterization of Alkalophilic Cellulase from the Symbiotic Bacillus subtilis BC1 of the Leopard Moth, Zeuzera pyrina (L.) (Lepidoptera: Cossidae)

• Published in: Current microbiology 2020-07, Vol.77 (7), p.1254-1261
• Main Authors: Dehghanikhah, Fahimeh, Shakarami, Jahanshir, Asoodeh, Ahmad
• Format: Article
• Language: English
• Subjects: Acetone; Animals; Bacillus subtilis; Bacillus subtilis - enzymology; Bacterial Proteins - chemistry; Bacterial Proteins - isolation & purification; Bacterial Proteins - metabolism; Biomedical and Life Sciences; Biotechnology; Butterflies & moths; Calcium chloride; Carboxymethyl cellulose; Carboxymethylcellulose; Cellobiose; Cellulase; Cellulase - chemistry; Cellulase - isolation & purification; Cellulase - metabolism; Cellulose; Endoglucanase; Enzyme Stability; Enzymes; Gel electrophoresis; Kinetics; Life Sciences; Metal ions; Microbiology; Moths - microbiology; Nickel chloride; Organic solvents; pH effects; Sodium lauryl sulfate; Solvents; Symbiosis; Thin layer chromatography; Zeuzera pyrina
• ISSN: 0343-8651; 1432-0991
• DOI: 10.1007/s00284-020-01938-z

In the current study, an extracellular cellulase belonging to symbiotic Bacillus subtilis Bc1 of the leopard moth is purified and characterized. The molecular mass of enzyme was 47.8 kDa using SDS-PAGE. The purified enzyme had optimum activity in temperature and pH around 60 °C and 8, respectively. The purified cellulase was introduced as a stable enzyme in a wide variety of temperature (20–80 °C) and pH (4–10) and remained active to more than 74% at 80 °C for 1 h. Moreover, the cellulase extremely was stabled in the presence of metal ions and organic solvents and its activity was increased by acetone (20% v/v), CaCl 2 and CoCl 2 and inhibited by MnCl 2 and NiCl 2 . The values of enzyme's K m and V max were found to be 1.243 mg/mL and 271.3 µg/mL/min, respectively. The purified cellulase hydrolyzed cellulose, avicel and carboxymethyl cellulose (CMC) and the final product of CMC hydrolysis was cellobiose using thin-layer chromatography analysis. Consequently, owing to exo/endoglucanase activity and organic solvent, temperature and pH stability of the purified cellulase belong to B. subtilis BC1, it can be properly employed for various industrial purposes.