I formed chitosan hydrogel using 2% acetic acid. Prior to bead formation, 3g of NaOH in 40 ml absolute ethanol was used as a coagulant and the hydrogel was dropped into the coagulant solution using a syringe. Spherical and smooth beads were formed. These beads were dried at 70 C for 48 hr in an oven. On optimizing immobilization by adsorption, 2 ml of the enzyme solution was introduced into a test tube containing 3 pieces of beads. The beads got dissolved in the enzyme solution.....WHAT DO I DO NOW TO SALVAGE THE SITUATION?
Consider the composition of the enzyme solution. What was its pH? Was it pH-buffered? Did it contain a chelator (such as EDTA) that might have bound up any divalent cations used in the coagulant?
You might be able to remedy the problem by dialyzing the enzyme solution to change its composition to one that is compatible with retaining the beads' structure. Also, concentrating the enzyme solution by ultrafiltration, so that a smaller volume of it must be added to the beads, could reduce the problem by reducing the amount of interfering substances added along with the protein.
Hi Ozoemena,
One thing you haven't mentioned in your question is the composition of the 'enzyme solution'. Did it contain polysaccharidase by any chance?