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

Microbial Cell Factories

Jenny Arnling Bååth¹  Peter Westh¹  Morten H. H. Nørholm²  Kenneth Jensen³  Sophia A. H. Heyde⁴ 

[1] Department of Biotechnology and Biomedicine, Technical University of Denmark, Søltofts Plads, 2800, Kgs. Lyngby, Denmark;Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet B220, 2800, Kgs. Lyngby, Denmark;Novozymes A/S, Biologiens Vej 2, 2800, Kgs. Lyngby, Denmark;Novozymes A/S, Biologiens Vej 2, 2800, Kgs. Lyngby, Denmark;Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet B220, 2800, Kgs. Lyngby, Denmark;

关键词: PETase;    Ideonella sakaiensis;    Surface display;    Extracellular protein production;    E. coli;   

DOI  :  10.1186/s12934-021-01582-7

来源: Springer

【 摘 要 】

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.

【 授权许可】

CC BY