Excretory expression of IsPETase in E. coli by an enhancer of signal peptides and enhanced PET hydrolysis.

The PET hydrolase from Ideonella sakaiensis (IsPETase) is efficient for PET degradation, which provides a promising solution for environmental contamination by plastics. This study focuses on improving the excretion of IsPETase from E. coli by signal peptide (SP) engineering. A SP enhancer B1 (MERACVAV) was fused to the N-terminal of commonly-used SP (PelB, MalE, LamB, and OmpA) to mediate excretion of IsPETase. Strikingly, the modified SP B1OmpA, B1PelB, and B1MalE significantly increased the excretion of IsPETase, while IsPETase was basically expressed in periplasmic space without enhancer B1. The excretion efficiency of IsPETase mediated by B1PelB was improved by 62 folds compared to that of PelB. The hydrolysis of PET by crude IsPETase in culture solution was also enhanced. Furthermore, the amount of released MHET/TPA from PET by IsPETase was increased by 2.7 folds with pre-incubation of hydrophobin HFBII. Taken together, this work may provide a feasible strategy for the excretion and application of the IsPETase.

3D printed smart silk wearable sensors.

Wearable sensors play a key role in point-of-care testing (POCT) for their flexible and integration capability for sensitive physiological and biochemical sensing. Here, we present a multifunction wearable silk patch with both electronic channels and microchannels by utilizing matrix-assisted sacrificial 3D printing methods. Owing to the unique properties of a composite silk film (polyvinyl alcohol (PVA) and silk fibroin (SF)), the wearable sensors possess excellent tensile properties, self-healing ability and biocompatibility. Multi-layer channel (microfluidics and microcircuit)-integrated silk wearable sensors were then fabricated for simultaneous sensitive sensing of human cancer markers (carcinoembryonic antigen (CEA) and alpha-fetoprotein (AFP)) and motion monitoring. These features of the silk wearable sensors indicate their potential value for sensitive sensing, which will enable them to find broader applications in many fields in POCT, artificial skin and organ-on-a-chip systems.

Excretory overexpression of hydrophobins as multifunctional biosurfactants in E. coli.

Hydrophobins are small amphipathic proteins excreted from filamentous fungi that self-assemble into the amphipathic film at hydrophobic/hydrophilic interfaces and can be used in a wide range of biotechnological application such as antimicrobial coatings, biosensors, and drug delivery. Here we describe a simple method for producing functionally active class I and class II hydrophobins in E. coli. The class I hydrophobin EAS (rodlet protein) from Neurospora crassa and class II hydrophobin HFBII from Trichoderma reesei were separately fused with fusion partner Ffu312 (ß-fructofuranosidase truncation with a native signal peptide) and successfully expressed in E. coli. Significantly, fused hydrophobins Ffu312-EAS and Ffu312-HFBII were excreted into the culture medium. The excretory expression of hydrophobins facilitated the correct disulfide-bond formation and simplified the purification. Both fusion hydrophobins reversed the glass surface hydrophilicity, reduced the water surface tension and improved emulsion stability. Ffu312 has little effect on surface coating, water surface tension and emulsion stabilization of hydrophobins. This study may provide an efficient approach for excretory and functional expression of class I and class II hydrophobins in E. coli.