Enhanced production of extracellular triacylglycerol lipase for bioplastic degradation by replacing signal peptide

Abstract

With the increase in plastic production, efficient and timely plastic degradation are urgently needed. In that point, biodegradable plastics have attracted attention as potential solutions for environmental pollution of plastics. However, finding of superior degrading strains and enzymes such as esterase, cutinase, and triacylglycerol lipase (TGL) of bioplastic are still needed together with the efficient secretion systems of degrading enzymes. As a result, we investigated methods to enhance protein expression and secretion of novel bioplastic degrading enzyme by using signal peptides. The genes encoding TGL from Bacillus sp. JY35 and various secretory (Sec) pathway signal peptides were cloned together by replacing the original signal sequence, and they were expressed under T7 promoters in Escherichia coli BL21 (DE3). Esterase activity with p-nitrophenol esters, a plate assay, and SDS-PAGE were performed to screen and evaluate signal peptide efficiency. As a result, the PhoA-TGL combination was the most effective against bioplastic degradation, achieving a Polycaprolactone (PCL) degradation efficiency of 77？%, which was approximately 3.3 times higher than that of TGL with the original signal peptide. Furthermore, Polybutylene succinate (PBS) degradation under similar conditions was 1.5 times higher. Overall, this study showed signal peptide engineering could enhance the extracellular secretion and degradation system of triacylglycerol lipase (TGL) and highlights the potential of PhoA signal peptides and E. coli host to enhance production and secretion of plastic-degrading enzyme and degrading system.