Application of a novel fluorogenic polyurethane analogue probe in polyester‐degrading microorganisms screening by microfluidic droplet

Abstract

Application of polyester‐degrading microorganisms or enzymes should be considered as an eco‐friendly alternative to chemical recycling due to the huge plastic waste disposal nowadays. However, current impranil DLN‐based screening of polyester‐degrading microorganisms is time‐consuming, labour‐intensive and unable to distinguish polyesterases from other protease‐ or amidase‐like enzymes. Herein, we present an approach that combined a novel synthetic fluorescent polyurethane analogue probe (FPAP), along with the droplet‐based microfluidics to screen polyurethane‐degrading microorganisms through fluorescence‐activated droplet sorting (FADS) pipeline. The fluorescent probe FPAP exhibited a fluorescence enhancement effect once hydrolysed by polyesterases, along with a strong specificity in discriminating polyesterases from other non‐active enzymes. Application of FPAP in a microfluidic droplet system demonstrated that this probe exhibited high sensitivity and efficiency in selecting positive droplets containing leaf‐branch compost cutinase (LCC) enzymes. This novel fluorogenic probe, FPAP, combined with the droplet microfluidic system has the potential to be used in the exploitation of novel PUR‐biocatalysts for biotechnological and environmental applications.

Improving plastic degrading enzymes via directed evolution

“…Methods such as FACS and FADS show significant potential for the selection of desired plastic-degrading enzymes from large libraries ( Qiao et al. 2022 , Xu et al. 2023 , Cribari et al.…”

Microbial Enzyme Biotechnology to Reach Plastic Waste Circularity: Current Status, Problems and Perspectives

“…The main bottleneck of this approach (function-based) is the availability of efficient and fast screening methods, as recently developed for PET [ 224 , 225 , 226 ]. Therefore, novel high-throughput screening methods suitable for enzymes and microorganisms must be developed for other polyesters, including polyester-based PURs, as shown by Xu et al [ 227 ].…”

Current trends in waste valorization

“…The authors centred their review and analysis on the following key points: (i) the physical, mechanical and chemical degradation (weathering) of plastics, microbial plastic colonization, (ii) the role of additional C sources for microbial degradation of plastics, (iii) the common structural traits of polyethylene terephthalate (PET)‐active enzymes, (iv) the model for enzymatic degradation of polyethylene terephthalate (PET) by mixed and multispecies microbial consortia and (v) the future challenges in this field. In this special issue, another work (Xu et al, 2022 ) has also examined the role of microorganisms in plastic degradation, and its authors have proven the use of a novel fluorogenic probe and fluorescence‐activated droplet sorting (FADS) pipeline for screening polyester‐degrading microorganisms. The authors assessed the specificity and sensitivity of the fluorogenic probe FPAP (fluorescent polyurethane analogue probe) for fluorescence detection in picolitre droplets.…”