4.7 Article

Surface display as a functional screening platform for detecting enzymes active on PET

Journal

MICROBIAL CELL FACTORIES
Volume 20, Issue 1, Pages -

Publisher

BMC
DOI: 10.1186/s12934-021-01582-7

Keywords

PETase; Ideonella sakaiensis; Surface display; Extracellular protein production; E; coli

Funding

  1. European Union's Horizon 2020 research and innovation program [887711]
  2. Novo Nordisk Foundation [NNFSA170028392, NNF17OC0027752]

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PET is the most common polyester plastic in the world, but its accumulation in nature is causing global environmental issues. Current recycling processes deteriorate the mechanical properties of PET, necessitating more efficient and bio-sustainable hydrolysis methods. A recently discovered PETase enzyme has shown potential for biodegrading PET, but engineering more efficient PETases is currently progressing slowly.
Poly(ethylene terephthalate) (PET) is the world's most abundant polyester plastic, and its ongoing accumulation in nature is causing a global environmental problem. Currently, the main recycling processes utilize thermomechanical or chemical means, resulting in the deterioration of the mechanical properties of PET. Consequently, polluting de novo synthesis remains preferred, creating the need for more efficient and bio-sustainable ways to hydrolyze the polymer. Recently, a PETase enzyme from the bacterium Ideonella sakaiensis was shown to facilitate PET biodegradation, albeit at slow rate. Engineering of more efficient PETases is required for industrial relevance, but progress is currently hampered by the dependency on intracellular expression in Escherichia coli. To create a more efficient screening platform in E. coli, we explore different surface display anchors for fast and easy assaying of PETase activity. We show that PETases can be functionally displayed on the bacterial cell surface, enabling screening of enzyme activity on PET microparticles - both while anchored to the cell and following solubilization of the enzymes.

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