Promote the expression and corrected folding of an extremely stable N-demethylase by promoter reconstruction, native environment simulation and surface design.

Thermomicrobium roseum sarcosine oxidase (TrSOX) was a N-demethylase with specific substrate chiral selectivity, outstanding thermostability and environmental resistance. To promote the expression of TrSOX in Bacillus subtilis W600, the HpaII promoter of pMA5 plasmid was replaced by constitutive or inducible promoters. Through orthogonal experiment, the expression process was optimized, B. subtilis W600 cells containing pMA5-Pxyl-trSOX plasmid were cultivated until OD600nm reached 2.0 and were then induced with 1.6% xylose at 37 °C for 2 h, and the native environment of T. roseum was simulated by heating at 80 °C, with the productivity of TrSOX increased from ~8.3 to ~66.7 µg/g wet cells; and the simulated high temperature was the key switch for the final folding. To reduce the surface hydrophobicity, a S320R mutant was built to form a hydrophilic lid around the entrance of the substrate pocket, and the yield of TrSOX (S320R) was ~163.0 µg/g wet cells, approximately 20 folds as that in the initial expression system. This mutant revealed the similar secondary structure, stability, resistance, chiral substrate selectivity and optimal reaction environment with wild type TrSOX; however, the N-demethylation activities for amino acid derivative substrates were dramatically increased, while those for hydrophobic non-amino acid compounds were repressed.

Interfacial Built-In Electric Field-Driven Direct Current Generator Based on Dynamic Silicon Homojunction.

Searching for light and miniaturized functional device structures for sustainable energy gathering from the environment is the focus of energy society with the development of the internet of things. The proposal of a dynamic heterojunction-based direct current generator builds up new platforms for developing in situ energy. However, the requirement of different semiconductors in dynamic heterojunction is too complex to wide applications, generating energy loss for crystal structure mismatch. Herein, dynamic homojunction generators are explored, with the same semiconductor and majority carrier type. Systematic experiments reveal that the majority of carrier directional separation originates from the breaking symmetry between carrier distribution, leading to the rebounding effect of carriers by the interfacial electric field. Strikingly, NN Si homojunction with different Fermi levels can also output the electricity with higher current density than PP/PN homojunction, attributing to higher carrier mobility. The current density is as high as 214.0 A/m2, and internal impedance is as low as 3.6 kΩ, matching well with the impedance of electron components. Furthermore, the N-i-N structure is explored, whose output voltage can be further improved to 1.3 V in the case of the N-Si/Al2O3/N-Si structure, attributing to the enhanced interfacial barrier. This approach provides a simple and feasible way of converting low-frequency disordered mechanical motion into electricity.

Tunable Dynamic Black Phosphorus/Insulator/Si Heterojunction Direct-Current Generator Based on the Hot Electron Transport.

Static heterojunction-based electronic devices have been widely applied because carrier dynamic processes between semiconductors can be designed through band gap engineering. Herein, we demonstrate a tunable direct-current generator based on the dynamic heterojunction, whose mechanism is based on breaking the symmetry of drift and diffusion currents and rebounding hot carrier transport in dynamic heterojunctions. Furthermore, the output voltage can be delicately adjusted and enhanced with the interface energy level engineering of inserting dielectric layers. Under the ultrahigh interface electric field, hot electrons will still transfer across the interface through the tunneling and hopping effect. In particular, the intrinsic anisotropy of black phosphorus arising from the lattice structure produces extraordinary electronic, transport, and mechanical properties exploited in our dynamic heterojunction generator. Herein, the voltage of 6.1 V, current density of 124.0 A/m2, power density of 201.0 W/m2, and energy-conversion efficiency of 31.4% have been achieved based on the dynamic black phosphorus/AlN/Si heterojunction, which can be used to directly and synchronously light up light-emitting diodes. This direct-current generator has the potential to convert ubiquitous mechanical energy into electric energy and is a promising candidate for novel portable and miniaturized power sources in the in situ energy acquisition field.

Cobalt sulfide aerogel prepared by anion exchange method with enhanced pseudocapacitive and water oxidation performances.

This work introduces the anion exchange method into the sol-gel process for the first time to prepare a metal sulfide aerogel. A porous Co9S8 aerogel with a high surface area (274.2 m2 g-1) and large pore volume (0.87 cm3 g-1) has been successfully prepared by exchanging cobalt citrate wet gel in thioacetamide and subsequently drying in supercritical ethanol. Such a Co9S8 aerogel shows enhanced supercapacitive performance and catalytic activity toward oxygen evolution reaction (OER) compared to its oxide aerogel counterpart. High specific capacitance (950 F g-1 at 1 A g-1), good rate capability (74.3% capacitance retention from 1 to 20 A g-1) and low onset overpotential for OER (220 mV) were observed. The results demonstrated here have implications in preparing various sulfide chalcogels.

[Membrane transfer-based colorimetric DNA detection using enzyme modified gold nanoparticles].

We report here a novel membrane transfer-based DNA detection method, in which alkaline phosphatase labeled gold nanoparticle (AuNP) probes were used as a means to amplify the detection signal. In this method, the capture probe P1, complimentary to the 3' end of target DNA, was immobilized on the chip. The multi-component AuNP probes were prepared by co-coating AuNPs with the detecting probe P2, complimentary to the 5' end of target DNA, and two biotin-labeled signal probes (T10 and T40) with different lengths. In the presence of target DNA, DNA hybridization led to the attachment of AuNPs on the chip surface where specific DNA sequences were located in a "sandwich" format. Alkaline phosphatase was then introduced to the surface via biotine-streptavidin interaction. By using BCIP/NBT alkaline phosphatase color development kit, a colorimetric DNA detection was achieved through membrane transfer. The signal on the membrane was then detected by the naked eye or an ordinary optical scanner. The method provided a detection of limit of 1 pmol/L for synthesized target DNA and 0.23 pmol/L for PCR products of Mycobacterium tuberculosis 16S rDNA when the ratio of probes used was 9:1:1 (T10:T40:P2). The method described here has many desirable advantages including high sensitivity, simple operation, and no need of sophisticated equipment. The method can be potentially used for reliable biosensings.