Purification and characterization of chitinase produced by thermophilic fungi Thermomyces lanuginosus

Background In the past few years, the production of shrimp shell waste from the seafood processing industries has confronted a significant surge. Furthermore, insignificant dumping of waste has dangerous effects on both nature and human well-being. This marine waste contains a huge quantity of chitin which has several applications in different fields. The chitinase enzyme can achieve degradation of chitin, and the chitin itself can be used as the substrate as well for production of chitinase. In the current study, the chitinase enzyme was produced by Thermomyces lanuginosus. The extracellular chitinase was purified from crude extract using ammonium sulfate precipitation followed by DEAE-cellulose ion-exchange chromatography and Sephadex G-100 gel filtration chromatography. The stability and activity of chitinase with different pH, temperature, different times for a reaction, in the presence of different metal ions, and different concentration of enzyme and substrate were analyzed. Result The chitinase activity was found to be highest at pH 6.5, 50 degrees C, and 60 min after the reaction began. and the chitinase showed the highest activity and stability in the presence of beta-mercaptoethanol (ME). The SDS-PAGE of denatured purified chitinase showed a protein band of 18 kDa. Conclusion The characterization study concludes that Cu2+, Hg2+, and EDTA have an inhibitory effect on chitinase activity, whereas beta-ME acts as an activator for chitinase activity. The utilization of chitin to produce chitinase and the degradation of chitin using that chitinase enzyme would be an opportunity for bioremediation of shrimp shell waste.

Automated summary

The research found that the chitinase enzyme showed the highest activity at pH 6.5, 50 degrees Celsius, and 60 minutes into the reaction. The enzyme also exhibited the highest activity and stability in the presence of beta-mercaptoethanol (ME). The denatured purified chitinase showed a protein band of 18 kDa on SDS-PAGE.