Catalytically Active Inclusion Bodies of Recombinant LipAMS8 lipase from Antarctic Pseudomonas sp

Heterologous expression of some recombinant protein in E. coli. triggered the formation of inclusion bodies. Recent studies revealed that the aggregated proteins formed are folded correctly and retain their native-like structure, hence possessing catalytic activity. LipAMS8 lipase originally isolated from Antarctic Pseudomonas was overexpressed in E. coli. BL21(De3)/pET32b and resulted in the formation of inclusion bodies. The inclusion bodies were isolated using mild solubilising agents, 50 mM Tris-HCl, 50 mM NaCl, and 1% Triton-X 100 (pH 8.0). Scanning electron microscopy (SEM) analysis depicted the particles as rod-like structures ranging from 1 µm to 100 nm. Biochemical characterisation of the inclusion bodies showcased their catalytic activity, optimum temperature, pH stability, metal ion interaction, and high tolerance to organic solvents. Retaining significant residual activity up to 50% over a range of pH and temperature. Storage stability of LipAMS8 CatIBs at 4 0 C and 25 0 C revealed that it could retain more than 50% of its activity for up to thirteen and eight weeks respectively. This indicated the novelty of the inclusion bodies of LipAMS8 lipase, which is expressed naturally and displays outstanding properties of high activity, pH stability, and extreme tolerance to organic solvents. LipAMS8 lipase CatIBs are catalytically active inclusion bodies that occur naturally as the recombinant enzyme in heterologous expression and can influence the production of cold-active lipase which is highly demanded in various industries for their production processes.