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Cefotaximase-Munich (CTX-M) extended-spectrum beta-lactamases (ESBLs) are commonly associated with Gram-negative, hospital-acquired infections worldwide. Several beta-lactamase inhibitors, such as clavulanate, are used to inhibit the activity of these enzymes. To understand the mechanism of CTX-M-15 activity, we have determined the crystal structures of CTX-M-15 in complex with two specific classes of beta-lactam compounds, desfuroylceftiofur (DFC) and ampicillin, and an inhibitor, clavulanic acid. The crystal structures revealed that Ser70 and five other residues (Lys73, Tyr105, Glu166, Ser130, and Ser237) participate in catalysis and binding of those compounds. Based on analysis of steady-state kinetics, thermodynamic data, and molecular docking to both wild-type and S70A mutant structures, we determined that CTX-M-15 has a similar affinity for all beta-lactam compounds (ceftiofur, nitrocefin, DFC, and ampicillin), but with lower affinity for clavulanic acid. A catalytic mechanism for tested β-lactams and two-step inhibition mechanism of clavulanic acid were proposed. CTX-M-15 showed a higher activity toward DFC and nitrocefin, but significantly lower activity toward ampicillin and ceftiofur. The interaction between CTX-M-15 and both ampicillin and ceftiofur displayed a higher entropic but lower enthalpic effect, compared with DFC and nitrocefin. DFC, a metabolite of ceftiofur, displayed lower entropy and higher enthalpy than ceftiofur. This finding suggests that compounds containing amine moiety (e.g., ampicillin) and the furfural moiety (e.g., ceftiofur) could hinder the hydrolytic activity of CTX-M-15. 

The majority of optical lenses have spherical surface profiles because they are convenient to fabricate. Replacing spherical optics with aspheric optics leads to smaller size, lighter weight, and less complicated optical systems with a superior imaging quality. However, fabrication of aspheric lenses is expensive and time-consuming. Here, we introduce a straightforward and low-cost casting method to fabricate polymeric aspheric lenses. An elastomeric ferrogel was formed into an aspherical profile by using a designed magnetic field and then was used as a mold. Different types of aspherical profiles from parabola to hyperbola can be formed with this method by tuning the magnetic field. A home-built Shack–Hartmann sensor was employed to characterize the cast polymeric lenses. The effects of magnetic field intensity, gradient of the magnetic field, and magnetic susceptibility of the ferrogel on the lens profiles were investigated. This technique can be used for rapid-forming polymeric aspherical lenses with different sizes and shapes.