Purification and characterization of cellulase from May beetle (Phyllophaga errans) gut
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Abstract. Ezima EN, Adegbesan BO, Osonuga IO, Adefuye AO, Adepoju AR, Bello TH, Olalekan SO. 2024. Purification and characterization of cellulase from May beetle (Phyllophaga errans) gut. Asian J Trop Biotechnol 21: 52-58. Cellulose constitutes a significant portion of plants' primary and secondary cell walls. Insects, traditionally believed to lack the ability to digest cellulose, have evolved distinctive strategies for cellulose degradation. May beetle (Phyllophaga errans LeConte, 1859) is herbivorous insects that feed on numerous plant species, including vegetables, maize plants, and other fresh food crops, causing a lot of damage to farm products. The ability of these beetles to thrive on plant diet may be due to the presence of highly effective cellulose digesting enzyme (cellulase) in their gut which aids the breaking down of cellulose into glucose. Therefore, this study focused on the extraction and purification of cellulase from the P. errans gut sourced from a farm in Ijebu-Ode, Ogun State, Nigeria. Following the dissection and homogenization of the beetle's guts, cellulase isolation and purification were carried out utilizing techniques such as ammonium sulfate precipitation, ion-exchange chromatography on CM-Sephadex G-200 and gel filtration using Sephacryl S-200 gel. The resulting pure cellulase from the gut of the P. errans exhibited a purification fold of 43.71 and a yield of 21.61% with a specific activity of 70.38 units/mg of protein. Characterization revealed the enzyme's native molecular weight of 24.6 kDa, Km (0.67 mM), and Vmax (192.31 mg/mL/min), with optimal activity observed at pH 9 and 70°C. The cellulase from the P. errans gut showcased distinctive characteristics that, if properly harnessed, could pave the way for practical applications in various industries, particularly in developing an efficient pesticide for pest management and environmental conservation.
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