An efficient strategy to tailor PET hydrolase: Simple preparation with high yield and enhanced hydrolysis to micro-nano plastics

Polyethylene terephthalate (PET) nano/microplastics (PET-NMPs) are regarded as an emergent hazardous waste for the environment. Enzymatic treatment of PET-NMPs is one of the most promising methods. However, strategies for mining or engineering of PET hydrolases with better characteristics and the simple and cost-effective preparation of them are the bottlenecks currently. Herein, we proposed a gene fusion strategy to tailor PET hydrolase (ICCG) with ferritin (namely F-C) towards micro-nano PET degradation. The purified F-C was obtained by an easy scalable low-speed centrifugation with 80.8 % activity recovery and 82.9 % protein recovery compared to the crude protein extraction, with the final high yield of 2.17 g/L. Encouragingly, unlike only hydrolyzing amorphous PET (crystallinity lower than 10 %), the resulted F-C showed 84.53 mg/h/mg specific activity at 70 °C for 5 h towards micro-PET with relatively high crystallinity (20.54 %) at the optimized enzyme/PET ratio of 1:100 (Wt), without producing intermediates. The supreme activity of F-C was closely related to its enhanced affinity towards substrate, increased substrate's ester bond tensions and binding pocket volume. More interestingly, F-C exhibited promising stability not only in storage or high temperature, but also in simulated seawater (hypersaline environment), with the half-lives of 128.4 days at 30 °C. Thus, the all-in-one strategy will offer a green and alternative solution to assist the PET-NMPs waste treatments such as recycling in the high-temperature reactor or degradation in seawater.