Immobilization of cellulase on magnetoresponsive graphene nano-supports

In this study, we report the preparation of pH tunable, temperature sensitive magnetoresponsive graphene-based nano-bio carriers for cellulase immobilization. We discuss a simple route to overcome the geometric disadvantage imposed by most 2D immobilization supports and make them capable of closely mimicking free enzymes (FE) operating under similar reaction conditions. The supramolecular assembly of oppositely charged quenched polyelectrolytes and maghemite–magnetite nanoparticles on 2D graphene supports followed by covalent immobilization of cellulase shows a marked improvement in the bio-receptivity of graphene supports. The incorporation of magnetic nanoparticles opens up the possibility of recovery and reuse of the enzyme over multiple cycles. The immobilized enzymes retained about 55% of the original specific activity even after four cycles of reuse. Cellulase immobilization is achieved by a combination of annealed polyelectrolyte brushes and zero-length spacer molecules. The swelling behavior of annealed polyelectrolyte brushes is a strong function of the environmental conditions. The degree of polyelectrolyte swelling can be easily tweaked by manipulating the pH and temperature, providing us an effective tool to control the activity of immobilized enzymes. At a pH of 5.1 and a temperature of 50 °C, the immobilized enzymes with the annealed polyelectrolyte brushes displayed close to 1.5-fold improvement in the activity as compared to immobilized enzymes without the brushes. Activity of immobilized cellulase is evaluated using both soluble as well as insoluble substrates like 2% (w/v) CMC and avicel respectively.

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► Immobilization of cellulase on 2D graphene supports using a combination of quenched and annealed polyelectrolytes is reported.
► Use of weak polyacrylic acid (PAA) brushes boosts the enzymatic activity of the immobilized cellulase helping overcome limitations associated with using flat 2D supports.
► Cellulase activity is a function of PAA brush swelling and can be adjusted by changing the pH and temperature.
► The use of magnetic nanoparticles in the assembly allows for recycle and reuse over multiple cycles reducing the cost of using fresh enzymes instead.