A separable and highly-stable enzyme system was developed by adsorption of phenylalanine ammonia lyase (PAL) from Rhodotorula glutinis in amino-functionalized macroporous silica gel and subsequent enzyme crosslinking. This resulted in the formation of cross-linked enzyme aggregates (PAL-CLEAs) into macroporous silica gel (MSG-CLEAs). The effect of adsorptive conditions, type of aggregating agent, its concentration as well as that of cross-linking agent was studied. MSG-CLEAs production was most effective using ammonium sulfate (40%-saturation), followed by cross-linking for 1 h with 1.5% (v/v) glutaraldehyde. The resulting MSG-CLEAs extended the optimal temperature and pH range compared to free PAL and PAL-CLEAs. Moreover, MSG-CLEAs exhibited the excellent stability of the enzyme against various deactivating conditions such as temperature and denaturants, and showed higher storage stability compared to the free PAL and the conventional PAL-CLEAs. Such as, after 6 h incubation at 60°C, the MSG-CLEAs still retained more than 47% of the initial activity whereas PAL-CLEAs only retained 7% of the initial activity. Especially, the MSG-CLEAs exhibited good reusability due to its suitable size and active properties. These results indicated that PAL-CLEAs on MSG might be used as a feasible and efficient solution for improving properties of immobilized enzyme in industrial application.

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Titanium and zirconia (ZrO2) ceramics are widely used in biomedical fields. This study aims to achieve reliable brazed joints of titanium/ZrO2 using biocompatible Au filler for implantable medical products. The effects of brazing temperature and holding time on the interfacial microstructures and mechanical properties of titanium/Au/ZrO2 joints were fully investigated by scanning electron microscopy (SEM), energy-dispersive spectrometry (EDS) and X-ray diffraction (XRD). The results indicated that the typical interfacial microstructure of the titanium/Au/ZrO2 joint was titanium/Ti3Au layer/TiAu layer/TiAu2 layer/TiAu4 layer/TiO layer/ZrO2 ceramic. With an increasing brazing temperature or holding time, the thickness of the Ti3Au + TiAu + TiAu2 layer increased gradually. The growth of the TiO layer was observed, which promoted metallurgical bonding between the filler metal and ZrO2 ceramic. The optimal shear strength of ~35.0 MPa was obtained at 1150 °C for 10 min. SEM characterization revealed that cracks initiated and propagated along the interface of TiAu2 and TiAu4 reaction layers.