I am doing immobilization of amylase from sodium alginate, beads does not form properly and they are not become hard or melted when separated from calcium chloride solution. please guide me how can i improve my immobilization protocol.
An article on how to is linked here.
https://pdfs.semanticscholar.org/74ed/71151912dd8c5b050ce47a8c84bab29ed7ac.pdf
"From the enzyme kinetic study, decrease in substrate affinity and velocity of enzyme reaction were observed after immoilization of enzyme. Immobilized α-amylase retained 35% activity after 10 reuses with 30 min of each reaction time."
If interested see my most recent publications and thesis. I went a different direction with both amylase and cellulase systems where I used cation polyelectrolytes to increase the enzyme kinetics.
You can find an answerfor your question in this reference:
Article The use of encapsulation as a proposed solution to avoid pro...
Add 3-4% solution of sodium alginate prepared in 0.1 M phosphate (sodium) buffer (pH 7) by warming at 50 deg C. Allow to cool to room temperature, then add 1ml of your sterile enzyme stock solution earlier mixed with 9 ml of sodium alginate solution. Suspend drop wise into pre-chilled calcium chloride solution of concentrations say 0.1, 0.2, 0.3, 0.4 or 0.5 M. Gently stir mixture at 4°C for 2 h. Recovered beads formed by filtration and wash with distilled water. Store beads in 0.1 M phosphate (sodium) buffer (pH 7.0) at 4°C.
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