Application of Directed Evolution and Machine Learning to Enhance the Diastereoselectivity of Ketoreductase for Dihydrotetrabenazine Synthesis.

Biocatalysis is an effective approach for producing chiral drug intermediates that are often difficult to synthesize using traditional chemical methods. A time-efficient strategy is required to accelerate the directed evolution process to achieve the desired enzyme function. In this research, we evaluated machine learning-assisted directed evolution as a potential approach for enzyme engineering, using a moderately diastereoselective ketoreductase library as a model system. Machine learning-assisted directed evolution and traditional directed evolution methods were compared for reducing (±)-tetrabenazine to dihydrotetrabenazine via kinetic resolution facilitated by BsSDR10, a short-chain dehydrogenase/reductase from Bacillus subtilis. Both methods successfully identified variants with significantly improved diastereoselectivity for each isomer of dihydrotetrabenazine. Furthermore, the preparation of (2S,3S,11bS)-dihydrotetrabenazine has been successfully scaled up, with an isolated yield of 40.7% and a diastereoselectivity of 91.3%.

Asymmetric Synthesis of ß-Hydroxyphosphonates via a Chemoenzymatic Cascade.

Chiral ß-hydroxyphosphonates are essential building blocks for organophosphorus compounds. However, the asymmetric synthesis of these units remains a significant challenge. Herein, we describe a one-pot chemoenzymatic cascade process to access chiral ß-hydroxyphosphonates, which combines photo-oxidative chemical reactions and bioreductions. The incorporation of photooxidation in the chemical reaction resulted in up to 92% yield and >99% enantiomeric excess (ee) of ß-hydroxyphosphonates in the cascade. In addition, the scale-up of diethyl (S)-(2-hydroxy-2-phenylethyl)phosphonate demonstrates the potential application of this strategy.

Surface Flashover Induced by Metal Contaminants Adhered to Tri-Post Epoxy Insulators under Superimposed Direct and Lightning Impulse Voltages.

Metal contaminants can distort the surface electric field of the tri-post epoxy insulator and cause serious surface charge accumulation, significantly reducing the insulation performance of the insulator under the superimposed DC and lightning impulse voltage. In this paper, an experimental platform for charge accumulation and surface flashover of tri-post epoxy insulators under the superimposed DC and lightning impulse voltage was built, by surface point measurement and charge inversion calculation, the surface charge distribution characteristics of tri-post insulators with attached particles was experimentally explored and the influence law of attached metal particles on the charge accumulation was discussed. The results show that metal particles can cause a surge in the surface charge density of the tri-post epoxy insulator, forming bipolar charge spots whose polarity is opposite to that of the adjacent electrodes. The adsorbed metal dust can cause the polarity reversal of adjacent surface charges, forming a large-area unipolar charge spot. Moreover, the flashover voltage of a tri-post insulator under DC superimposed lightning impulse voltage with a clean surface and attached metal particle was measured, and the synergistic induction mechanism of charge spot accumulation and metal particle discharge on the flashover along the face of the tri-post insulator is thereby revealed. Compared with the clean insulators, the surface flashover voltages of tri-post epoxy insulators with metal contaminants adhered decrease under the superimposed voltages of different polarities, but the decline amplitude is greater under the heteropolar composite voltage. When adhered to the middle of the insulator leg, the distribution range of bipolar charge spots is the widest, and the surface flashover voltage decreases sharply, which can drop by 32% compared with the absence of particles. In addition, when the metal dust adsorbed by the tri-post epoxy insulator has a wide distribution range, the impact of metal dust on the flashover voltage is greater than that of the attached metal particles, and its hazard cannot be ignored. The research results can provide a reference for the insulation test method and optimal design of the DC tri-post epoxy insulator.

Frequency and Temperature-Dependent Space Charge Characteristics of a Solid Polymer under Unipolar Electrical Stresses of Different Waveforms.

In this paper, we studied the space charge phenomena of a solid polymer under thermal and electrical stresses with different frequencies and waveforms. By analyzing the parameter selection method of a protection capacitor and resistor, the newly built pulsed electro-acoustic (PEA) system can be used for special electrical stresses under 500 Hz, based on which the charge phenomena are studied in detail under positive and negative DC and half-wave sine and rectangular wave voltages. Experimental results show that the charge accumulated in the polyimide polymer under DC conditions mainly comes from the grounded electrode side, and the amount of charge accumulated with electric field distortion becomes larger in a high-temperature environment. At room temperature, positive charges tend to accumulate in low-frequency conditions under positive rectangular wave voltages, while they easily appear under high-frequency situations of negative ones. In contrast, the maximum electric field distortion and charge accumulation under both half-wave sine voltages occur at 10 Hz. When the measurement temperature increases, the accumulated positive charge decreases, with a more negative charge appearing under rectangular wave voltages, while a more positive charge accumulates at different frequencies of half-wave sine voltages. Therefore, our study of the charge characteristics under different voltage and temperature conditions can provide a reference for applications in the corresponding environments.