RESUMEN
Polyethylene terephthalate (PET) is a synthetic polymer consisting of ester bond-linked terephthalate and ethylene glycol. Tremendous amounts of PET have been produced and majority of them enters terrestrial and marine environment as wastes, posing serious threats to the global ecosystems. In 2016, a PET hydrolase from a PET-assimilating bacterium Ideonalla sakaiensis was reported and termed as IsPETase. This enzyme outperforms other PET-hydrolyzing enzymes in terms of its PET hydrolytic activity at ambient temperature, thus holds a great promise for PET biodegradation. In order to improve IsPETase activity, we conducted structure-based engineering to modify the putative substrate-binding tunnel. Among the several variants to the N233 residue of IsPETase, we discovered that the substitution of N233 with alanine increases its PET hydrolytic activity, which can be further enhanced when combined with a R280A mutation. We also determined the X-ray crystal structure of the IsPETase N233A variant, which shares nearly identical fold to the WT protein, except for an open end of subsite â ¡. We hypothesized that the smaller side chain of N233A variant might lead to an extended subsite â ¡ for PET binding, which subsequently increases the enzymatic activity. Thus, this study provides new clues for further structure-based engineering of PETase.
Asunto(s)
Burkholderiales , Hidrolasas , Tereftalatos Polietilenos/metabolismo , Burkholderiales/enzimología , Hidrolasas/genética , Ingeniería de ProteínasRESUMEN
Uranium enrichment measurement is an essential quality inspection for fuel rods before delivery to users. Generally, compared with active neutron assay (ANA) equipment, passive gamma-ray assay (PGA) equipment is more economical and safer. However, the current PGA equipment based on photomultipliers is too slow (1 m/min) to meet the growing needs in China. Recently, we have developed a set of compact high-speed PGA equipment including four detection modules (128 units in total), a 128-channel data acquisition system (DAS), a power supply, special software, and an automatic loading and unloading mechanism. The detection unit is based on silicon photomultipliers in virtue of its compact size and good performance. The DAS processes signals of all units in parallel into a sequence of data packets carrying the energy information and the corresponding unit ID. The software integrates the data packets into a fluctuating count curve in a time-delay superposition method and identifies possible abnormal pellets. After calibrations, our equipment can locate abnormal pellets accurately at a speed of 6 m/min. In addition, it can directly measure the enrichment of fresh pellets not in secular equilibrium without waiting for two months. So far, the equipment has been successfully run for one year on the assembly line of China North Nuclear Fuel Co. and shows good potential to replace the traditional ANA equipment.
RESUMEN
The scandium-catalyzed redox-neutral cascade [1,5]-hydride transfer/cyclization between C4-amine-substituted isatins and 1,3-dicarbonyl compounds has been developed. This protocol enabled the synthesis of tricyclic [3,4]-fused oxindoles in good to high yields and excellent diastereoselectivities, featuring high atom- and step economy as well as good functional group tolerance.
RESUMEN
The hydride transfer involved redox-neutral cascade cyclization has been developed to construct the spirocyclic bisoxindoles featuring a [3,4]-fused oxindole moiety from rationally designed C4-amine-substituted isatins, affording the diverse tricyclic [3,4]-fused oxindoles with three consecutive chiral centers in good yields and excellent diastereoselectivities (>20:1).
RESUMEN
An environmentally benign cascade redox-neutral process was developed for the efficient construction of pharmaceutically significant spirocyclic tetrahydro-quinolines via a 3-step cascade Knoevenagel condensation/[1,5]-hydride transfer/cyclization, which features green and additive-free conditions, wide substrate scope, and high step- and atom-economy.
