Team:DUT China/Description

Description

Description

Wide spreading and utilization of plastic polyethylene terephthalate (PET) in the world has caused a large quantity of environmental challenges and gained much attention. Because of its nonbiodegradability, a large amount of plastic waste accumulation forms a global environmental crisis [1]. Notably, in 2016, Yoshida et al. reported the discovery of the bacterium, Ideonella sakaiensis [2]. Microbe Ideonella sakaiensis was reported to be capable of secreting two efficient enzymes to deconstruct PET polymers in mild temperature. First, Ideonella sakaiensis PETase, a structurally well-characterized consensus α/β-hydrolase fold enzyme, converts PET to mono-(2-hydroxyethyl) terephthalate (MHET). MHETase, the second key enzyme, hydrolyzes MHET to the PET educts terephthalate and ethylene glycol [3].

However, this two-enzyme system degradation capacity is highly limited by inhibition effects, diffusion of intermediates and PET surface physicochemical properties [4]. Here, we design a delicate multicomplex enzyme system, in which short peptide tags (RIAD and RIDD) are applied to create scaffold-free modular enzymes assemblies [5]. In order to effectively degrade microplastic PET particles, we construct enzymes of IsPETase and MHETase and protein hydrophobin in our complex system via scaffold modular part, which reveal higher catalytic efficiency in mild temperature. All proteins involved have unique functions in our system. IsPETase and MHETase are two enzymes involved deconstructing polymer PET plastic to MHET and MHET to TPA molecules, respectively. And hydrophobin protein, a small fungal protein, possess positive effects on altering the physicochemical properties of PET surfaces and enzyme aggregation enhancement when it was fused with PET degradation enzyme cutinase [6]. In conclusion, our work presents an innovative strategy to improve PET degradation via biosynthetic factories and artificially designed proteins system that do not exist in the nature.

References

[1] Hachisuka Shin-ichi, Nishii Tarou, Yoshida Shosuke, & Cann Isaac. Development of a Targeted Gene Disruption System in the Poly(Ethylene Terephthalate)-Degrading Bacterium Ideonella sakaiensis and Its Applications to PETase and MHETase Genes. Applied and Environmental Microbiology 87, e00020-21.

[2] Yoshida S, Hiraga K, Takehana T, et al. A bacterium that degrades and assimilates poly(ethylene terephthalate).[J]. Science 351, 1196-1199. (2016).

[3] Palm, G. J., Reisky, L., Böttcher, D. et al. Structure of the plastic-degrading Ideonella sakaiensis MHETase bound to a substrate. Nature Communications 10, 1717 (2019).

[4] Knott, B. C. et al. Characterization and engineering of a two-enzyme system for plastics depolymerization. Proc Natl Acad Sci USA 117, 25476–25485 (2020).

[5] Kang, W. et al. Modular enzyme assembly for enhanced cascade biocatalysis and metabolic flux. Nature Communications 10, 4248 (2019).

[6] Ribitsch Doris et al. Enhanced Cutinase-Catalyzed Hydrolysis of Polyethylene Terephthalate by Covalent Fusion to Hydrophobins. Applied and Environmental Microbiology 81, 3586–3592 (2015).