Directed Evolution of an Efficient Polycarbonate Depolymerase With Exceptional Operational Stability
Abstract
ABSTRACT We recently developed a high‐throughput directed evolution platform for engineering polymer degrading enzymes, and showcased its utility through the development of an efficient and thermostable variant of Is PETase, termed HotPETase. Here, we show that this platform can be used to re‐engineer PET degrading enzymes for the recycling of other aromatic‐containing commodity polymers. Promiscuous poly(bisphenol‐A carbonate) (PC) depolymerase activity of LCC ICCG was enhanced by directed evolution to afford an engineered polycarbonate hydrolase, that also benefits from improved solvent tolerance and operational stability at elevated temperatures. Interestingly, the enzyme‐concentration dependent inhibition observed with the parent enzyme is also alleviated through evolution, improving practical utility. PC‐2 can achieve rapid and complete depolymerization of a PC film to bisphenol‐A (BPA) in just 6 h at 75°C. This study shows how plastic degrading enzymes can be readily adapted through evolution to operate on new and valuable polymer classes.
Article Details
Authors (8)
Henry A. Jones
Manchester Institute of Biotechnology, School of Chemistry The University of Manchester Manchester UK
Amy E. Hutton
Department of Chemistry, University of Manchester, Manchester Institute of Biotechnology, 131 Princess Street, Manchester M1 7DN, U.K.
Dominic Harris‐Jukes
Sustainable Materials Innovation Hub, Department of Materials University of Manchester Manchester UK
John Davidson
Manchester Institute of Biotechnology, School of Chemistry The University of Manchester Manchester UK
Linus O. Johannissen
Colin W. Levy
Department of Chemistry, University of Manchester, Manchester Institute of Biotechnology, 131 Princess Street, Manchester M1 7DN, U.K.
Michael P. Shaver
Sustainable Materials Innovation Hub, Department of Materials University of Manchester Manchester UK
Anthony P. Green