Fluorescence-activated droplet sorting of PET degrading microorganisms.

Enzymes that can decompose synthetic plastics such as polyethylene terephthalate (PET) are urgently needed. Still, a bottleneck remains due to a lack of techniques for detecting and sorting environmental microorganisms with vast diversity and abundance. Here, we developed a fluorescence-activated droplet sorting (FADS) pipeline for high-throughput screening of PET-degrading microorganisms or enzymes (PETases). The pipeline comprises three steps: generation and incubation of droplets encapsulating single cells, picoinjection of fluorescein dibenzoate (FDBz) as the fluorogenic probe, and screening of droplets to obtain PET-degrading cells. We characterized critical factors associated with this method, including specificity and sensitivity for discriminating PETase from other enzymes. We then optimized its performance and compatibility with environmental samples. The system was used to screen a wastewater sample from a PET textile mill. We successfully obtained PET-degrading species from nine different genera. Moreover, two putative PETases from isolates Kineococcus endophyticus Un-5 and Staphylococcus epidermidis Un-C2-8 were genetically derived, heterologously expressed, and preliminarily validated for PET-degrading activities. We speculate that the FADS pipeline can be widely adopted to discover new plastic-degrading microorganisms and enzymes in various environments and may be utilized in the directed evolution of degrading enzymes using synthetic biology.

Multifunctional cotton non-woven fabrics coated with silver nanoparticles and polymers for antibacterial, superhydrophobic and high performance microwave shielding.

Multifunctional cotton fabrics have attracted significant attention as next-generation wearable materials. Herein, we report a facile method for the fabrication of flexible and wearable cotton fabrics with ultra-high electromagnetic interference (EMI) shielding, antibacterial, and superhydrophobic properties. Cotton fabrics were first coated chemically with silver nanoparticles using polydopamine as adhesive and then with hydrophobic polydimethylsiloxane or polyimide. The introduction of polydopamine significantly increased the bond between silver nanoparticles and cotton fibers, thereby preventing silver nanoparticles from falling off the surface. The composite fabrics exhibited a high conductivity of ~1000 S/cm, and their EMI shielding effectiveness increased up to ~110 dB. The composite fabrics exhibited excellent self-cleaning performance and acid-alkali corrosion resistance because of their superhydrophobicity. Notably, the fabric composites showed a significant antibacterial action against Staphylococcus aureus and Escherichia coli.

Effect of organic solvent on enzymatic degradation of cyclic PBS-based polymers by lipase N435.

Poly(butylene succinate-co-cyclohexane dimethanol succinate) (P(BS-co-CHDMS)) and poly(butylene succinate-co-butanediol cyclohexanedicarboxylic acid) (P(BS-co-BCHDA)) were catalytically degraded by Candida antarctica lipase Novozyme 435 (N435) in CHCl3 and THF. The results indicated that the degradation rate was P(BS-co-CHDMS) > P(BS-co-BCHDA) > poly(butylene succinate) (PBS). The degradation rate of copolyesters was higher in CHCl3 than in THF, the highest degradation rate of 67% being obtained for P(BS-co-CHDMS). Hence, the CHCl3 solvent is more suitable for the enzyme-catalytic degradation of copolyesters, since the lipase can easier recognize the butylene succinate (BS-), (butanediol cyclohexanedicarboxylic acid) (BCA-), and (cyclohexane dimethanol succinate-type) (CMS-type) ester bonds in this solvent. Moreover, it can recognize the CMS-type ester bonds with a higher specificity than the (butanediol cyclohexanedicarboxylic acid type) (BCA-type) ester bonds. Molecular simulation results indicated that the structure of the lipase was stable in CHCl3 and THF. However, CHCl3 proved to be more suitable for a stable activity of the enzyme. The active pocket contains acyl-binding hydrophilic residues which are recognized by the substrate. The increase in the content of saturated cycles can increase the hydrophobicity of the substrate and thus, the amount of substrate bond to enzyme active site is increased, which facilitates the enzymatic degradation of copolyesters.

Influence of ether linkage on the enzymatic degradation of PBS copolymers: Comparative study on poly (butylene succinate-co-diethylene glycol succinate) and poly (butylene succinate-co-butylene diglycolic acid).

The difference of enzymatic degradation behavior between Poly (butylene succinate-co-diethylene glycol succinate) (PBS-co-DEGS) and Poly (butylene succinate-co-butylene diglycolic acid) (PBS-co-BDGA) was studied in a Tetrahydrofuran (THF)/toluene mixed system by Novozym 435 (N435, immobilized Candida Antarctica lipase supported on acrylic resin) catalysis for 30 h. These two copolymers (modified with alcoholic acid by ether linkage) were synthesized by melt polycondensation and characterized by 1H NMR. The average molecular weight and thermal property before and after degradation were determined by gel permeation chromatography (GPC) and thermogravimetric analysis (TGA), respectively. Results revealed that end-chain degradation of DEG20 (20% content diethylene glycol of diols) and intramolecular random degradation of DGA20 (20% content diglycolic acid of diacids) both occurred at the same time from 0 h to 12 h. TGA curves show that after degradation by N435, the T-5% of both copolymers decreased from about 300 °C to below 210 °C. In degradation products (linear and cyclic oligomers, no monomer was appeared below 10 degree of polymerization. According to the molecular docking results, the free binding energy between PC lipase and substrate was in the order of BDGAB < DEGSDEG < BSDEG < BSB. Thus, the enzymatic degradability of PBS-co-DEGS is more effective than that of PBS-co-BDGA.