RESUMO
The combination of wet-lab experimental data on multi-site combinatorial mutations and machine learning is an innovative method in protein engineering. In this study, we used an innovative sequence-activity relationship (innov'SAR) methodology based on novel descriptors and digital signal processing (DSP) to construct a predictive model. In this paper, 21 experimental (R)-selective amine transaminases from Aspergillus terreus (AT-ATA) were used as an input to predict higher thermostability mutants than those predicted using the existing data. We successfully improved the coefficient of determination (R2) of the model from 0.66 to 0.92. In addition, root-mean-squared deviation (RMSD), root-mean-squared fluctuation (RMSF), solvent accessible surface area (SASA), hydrogen bonds, and the radius of gyration were estimated based on molecular dynamics simulations, and the differences between the predicted mutants and the wild-type (WT) were analyzed. The successful application of the innov'SAR algorithm in improving the thermostability of AT-ATA may help in directed evolutionary screening and open up new avenues for protein engineering.
Assuntos
Aminoácidos , Engenharia de Proteínas , Aminoácidos/genética , Simulação de Dinâmica Molecular , Transaminases/metabolismo , Aprendizado de Máquina , Estabilidade EnzimáticaRESUMO
Meerwein-Ponndorf-Verley (MPV)-type reduction between benzonitriles and benzylic alcohols under transition-metal-free conditions has been demonstrated for the first time. Using simple KOt-Bu as the base, various benzonitriles can be efficiently reduced by benzylic alcohols via hydrogen transfer reduction, and the resultant phenyl imine can react further with benzylic alcohols to give amides as the final product in which both the alcohols and the nitriles are incorporated. Preliminary mechanistic investigations indicated that the reaction may go through multiple MPV-type hydrogen transfer processes.
RESUMO
An efficient method for direct trifluoromethylation and perfluoroalkylation at C3 of imidazopyridines through visible light-promoted C-H bond functionalization was developed. Under the irradiation of a blue LED, a series of C3-perfluoroalkyl-substituted imidazopyridines were synthesized from the corresponding imidazopyridines and perfluoroalkyl iodides in moderate to good yields at room temperature. It should be mentioned that this reaction proceeded in the absence of any transition-metal catalyst, oxidant and photocatalyst.
RESUMO
With the continuous progress of aerospace, military technology, and marine development, the MEMS resonance pressure sensor puts forward the requirements of not only a wide range but also high sensitivity. However, traditional resonators are hardly compatible with both. In response, we propose a new sensor structure. By arranging the resonant beam and the sensitive diaphragm vertically in space, the new structure improves the rigidity of the diaphragm without changing the thickness of the diaphragm and achieves the purpose of increasing the range without affecting the sensitivity. To find the optimal structural parameters for the sensor sensitivity and range, and to prevent the effects of modal disturbances, we propose a multi-objective optimization design scheme based on the BP and NSGA-II algorithms. The optimization of the structure parameters not only improved the sensitivity but also increased the interference frequency to solve the issue of mode interference. The optimized structure achieves a sensitivity and range of 4.23 Hz/kPa and 1-10 MPa, respectively. Its linear influence factor is 38.07, significantly higher than that of most resonant pressure sensors. The structural and algorithmic optimizations proposed in this paper provide a new method for designing resonant pressure sensors compatible with a wide range and high sensitivity.
RESUMO
Gelation, as an important functional property of soy protein isolate (SPI), can be improved by some green technologies in food manufacturing, including ultrasound, ultrahigh pressure and microwave treatments. This work investigated the effect of an alkaline solubilisation step in SPI extraction combined with sonication on protein properties. The TGase-induced gel of the modified SPI was prepared to explore the effect of ultrasound on gel properties, including structures, strength, water-holding capacity and rheological properties. Additionally, the differences between traditional ultrasound modification of SPI and current modification methods were analyzed. The results showed that the ultrasonication-assisted extraction method could result in a significant increase in extraction rate from 24.68% to 42.25%. Moreover, ultrasound-assisted modification of SPI gels induced with transglutaminase (TGase) exhibited significant improvement in mechanical properties, such as texture, water-holding capacity and rheological properties, In particular, SPI extracted at 400 W ultrasound intensity for 180 s showed the best overall performance in terms of gel properties. Our method efficiently uniformizes gel structure, enhancing mechanical properties compared to conventional ultrasound methods, which reduced energy consumption and costs. These findings provide insights into the production of high-gelation SPI in food manufacturing.
RESUMO
High performance sorting of circulating tumor cells (CTCs) from peripheral blood is key to liquid biopsies. Size-based deterministic lateral displacement (DLD) technique is widely used in cell sorting. But conventional microcolumns have poor fluid regulation ability, which limits the sorting performance of DLD. When the size difference between CTCs and leukocytes is small (e.g., less than 3 µm), not only DLD, many size-based separation techniques fail due to low specificity. CTCs have been confirmed to be softer than leukocytes, which could serve as a basis for sorting. In this study, we presented a multistage microfluidic CTCs sorting method, first sorting CTCs using a size-based two-array DLD chip, then purifying CTCs mixed by leukocytes using a stiffness-based cone channel chip, and finally identifying cell types using Raman techniques. The entire CTCs sorting and analysis process was label free, highly pure, high-throughput and efficient. The two-array DLD chip employed a droplet-shaped microcolumn (DMC) developed by optimization design rather than empirical design. Attributed to the excellent fluid regulation capability of DMC, the CTCs sorter system developed by parallelizing four DMC two-array DLD chips was able to process a sample of 2.5 mL per minute with a recovery efficiency of 96.30 ± 2.10% and a purity of 98.25 ± 2.48%. To isolate CTCs mixed dimensionally by leukocytes, a cone channel sorting method and chip were developed based on solid and hydrodynamic coupled analysis. The cone channel chip allowed CTCs to pass through the channel and entrap leukocytes, improving the purity of CTCs mixed by leukocytes by 1.8-fold.
Assuntos
Técnicas Biossensoriais , Técnicas Analíticas Microfluídicas , Células Neoplásicas Circulantes , Humanos , Microfluídica , Linhagem Celular Tumoral , Separação Celular/métodos , Células Neoplásicas Circulantes/patologiaRESUMO
Methylammonium lead triiodide perovskite (CH3NH3PbI3, MAPbI3) has been emerging as an easy processing and benign defect material for optoelectronic devices. Fiber-like perovskite materials are especially in demand for flexible applications. Here we report on a kind of polyacrylonitrile (PAN)/MAPbI3 composite fiber, which was electrospun from the mixing solution of PAN and MAPbI3. The absorption edge and optical gap of the PAN/MAPbI3 composite fibers can be easily tuned as the ratio of the perovskite changes. Both the moisture stability and the thermal stability of the perovskite are improved with the protection of PAN polymers. Flexible photodetectors based on this perovskite fiber were fabricated and analyzed. The photoresponse of the detector was highly sensitive to broadband visible light, and reached 6.5 µA W-1 at 700 nm with a voltage bias of 10 V. Compared with pure MAPbI3 photodetectors, this composite fiber photodetector has much-improved stability and flexibility, which can even be used to detect motion-related angular changes.