Biosensor and chemo-enzymatic one-pot cascade applications to detect and transform PET-derived terephthalic acid in living cells
Thomas Bayer,
Lara Pfaff,
Yannick Branson,
Aileen Becker,
Shuke Wu,
Uwe T. Bornscheuer,
Ren Wei
Affiliations
Thomas Bayer
Institute of Biochemistry, Department of Biotechnology & Enzyme Catalysis, University of Greifswald, Felix-Hausdorff-Straße 4, 17487 Greifswald, Germany; Institute of Molecular Biotechnology, TU Graz, Petersgasse 14, 8010 Graz, Austria; Corresponding author
Lara Pfaff
Institute of Biochemistry, Department of Biotechnology & Enzyme Catalysis, University of Greifswald, Felix-Hausdorff-Straße 4, 17487 Greifswald, Germany
Yannick Branson
Institute of Biochemistry, Department of Biotechnology & Enzyme Catalysis, University of Greifswald, Felix-Hausdorff-Straße 4, 17487 Greifswald, Germany
Aileen Becker
Institute of Biochemistry, Department of Biotechnology & Enzyme Catalysis, University of Greifswald, Felix-Hausdorff-Straße 4, 17487 Greifswald, Germany
Shuke Wu
Institute of Biochemistry, Department of Biotechnology & Enzyme Catalysis, University of Greifswald, Felix-Hausdorff-Straße 4, 17487 Greifswald, Germany; College of Life Science & Technology, Huazhong Agricultural University, Shizishan Street 1, Wuhan 430070, China
Uwe T. Bornscheuer
Institute of Biochemistry, Department of Biotechnology & Enzyme Catalysis, University of Greifswald, Felix-Hausdorff-Straße 4, 17487 Greifswald, Germany
Ren Wei
Institute of Biochemistry, Department of Biotechnology & Enzyme Catalysis, University of Greifswald, Felix-Hausdorff-Straße 4, 17487 Greifswald, Germany; Corresponding author
Summary: Plastic waste imposes a serious problem to the environment and society. Hence, strategies for a circular plastic economy are demanded. One strategy is the engineering of polyester hydrolases toward higher activity for the biotechnological recycling of polyethylene terephthalate (PET). To provide tools for the rapid characterization of PET hydrolases and the detection of degradation products like terephthalic acid (TPA), we coupled a carboxylic acid reductase (CAR) and the luciferase LuxAB. CAR converted TPA into the corresponding aldehydes in Escherichia coli, which yielded bioluminescence that not only semiquantitatively reflected amounts of TPA in hydrolysis samples but is suitable as a high-throughput screening assay to assess PET hydrolase activity. Furthermore, the CAR-catalyzed synthesis of terephthalaldehyde was combined with a reductive amination cascade in a one-pot setup yielding the corresponding diamine, suggesting a new strategy for the transformation of TPA as a product obtained from PET biodegradation.
