Construction and expression of gene coding MHETase in E. coli
17 viewsDOI:
https://doi.org/10.54939/1859-1043.j.mst.FEE.2024.280-285Keywords:
Plastic waste; MHETase; Plastic-degrading enzyme; Polyethylene terephthalate (PET); Recombinant protein.Abstract
Polyethylene terephthalate (PET) is a type of synthetic plastic widely used in many applications, especially packaging production. With the increasing production, consumption rates, and a short life cycle, the abundant amount of waste from PET plastic in the environment seriously affects human health as well as other living organisms. Currently, plastic treatment solutions that are safe, sustainable, and environmentally friendly have received much attention, especially biodegradation methods using natural microorganisms or enzymes for plastic treatment. To date, PETase and MHETase are two enzymes that have been studied with great potential in PET plastic degradation. In this study, we have successfully designed and expressed the gene encoding the enzyme MHETase using the E. coli BL21(DE3) expression system. The recombinant strain harboring the plasmid pET22b(+)-MHETase was cultured in LB medium containing 100 µg/mL Ampicillin, and extracellular protein was collected after 48 hours with the induction of 0.1 mM IPTG and cultivation at 30 °C. The results of the analysis by Western-blotting and SDS-PAGE protein electrophoresis showed that the expressed MHETase had a molecular weight of about 63 kDa, which is equivalent to the theoretical molecular weight. The specific activity of recombinant MHETase enzyme obtained by precipitation of (NH4)2SO4 salt was 85.88 U/mL. These results show the potential of MHETase enzyme for application in the degradation of PET plastic waste.
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