Laccase immobilized peroxidase mimicking magnetic metal organic frameworks for industrial dye degradation.

In the present work, laccase was successfully immobilized in peroxidase mimicking magnetic metal organic frameworks (MMOFs) within 30 min using a facile approach. The integration of magnetic nanoparticles during synthesis significantly eases the separation of prepared biocatalyst using an external magnet. The immobilization of laccase was confirmed using different characterization techniques. The laccase@MMOFs found spherical in nature with an average particle size below 100 nm. The synthesized laccase embedded framework exhibits supermagnetic property with the saturation magnetization (Ms) of 34.12 emu/gm. The prepared bio-metallic frameworks maintain high surface area and thermal stability. The laccase@MMOFs was successfully exploited for the degradation of industrial dyes in batch and continuous mode with an average degradation efficiency of 95%. The prepared laccase structure had an excellent recyclability retaining upto 89% residual activity upto 10th cycle and can be stored at room temperature upto 30 days without any significant loss of activity.

Ultrasonic hyperactivation of cellulase immobilized on magnetic nanoparticles.

In the present work, effect of low power, low frequency ultrasound on cellulase immobilized magnetic nanoparticles (cellulase@MNPs) was studied. To gain maximum activity recovery in cellulase@MNPs various parameters viz. ratio of MNPs:cellulase, concentration of glutaraldehyde and cross-linking time were optimized. The influence of ultrasonic power on cellulase@MNPs was studied. Under ultrasonic conditions at 24kHz, 6W power, and 6min of incubation time there was almost 3.6 fold increased in the catalytic activity of immobilized cellulase over the control. Results also indicated that there was improvement in pH and temperature stability of cellulase@MNPs. Furthermore, thermal deactivation energy required was more in cellulase@MNPs than that of the free cellulase. Secondary structural analysis revealed that there were conformational changes in free cellulase and cellulase@MNPs before and after sonication which might be responsible for enhanced activity after ultrasonication. Finally, the influence of ultrasound and cellulase@MNPs for biomass hydrolysis was studied.

Synthesis of glycinamides using protease immobilized magnetic nanoparticles.

In the present investigation, Bacillus subtilis was isolated from slaughterhouse waste and screened for the production of protease enzyme. The purified protease was successfully immobilized on magnetic nanoparticles (MNPs) and used for the synthesis of series of glycinamides. The binding and thermal stability of protease on MNPs was confirmed by FTIR spectroscopy and TGA analysis. The surface morphology of MNPs before and after protease immobilization was carried out using SEM analysis. XRD pattern revealed no phase change in MNPs after enzyme immobilization. The processing parameters for glycinamides synthesis viz. temperature, pH, and time were optimized using Response Surface Methodology (RSM) by using Design Expert (9.0.6.2). The maximum yield of various amides 2 butyramidoacetic acid (AMD-1,83.4%), 2-benzamidoacetic acid (AMD-2,80.5%) and 2,2'((carboxymethyl) amino)-2-oxoethyl)-2-hydroxysuccinyl)bis(azanediyl))diacetic acid (AMD-3,80.8%) formed was observed at pH-8, 50 °C and 30 min. The synthesized immobilized protease retained 70% of the initial activity even after 8 cycles of reuse.