Dear, Its really very difficult to measure the activity of immobilized cellulase as almost all subsrates for cellulase are insoluble and if you immobilize your enzyme your enzyme will not cleave the substrate. However, you can measure some of the activities of cellulase complex such as beta-glucosidase by performing your routine assay and including the beads in place of enzyme solution
I do not know why you have immobilized cellulase. You can determine carboxymethyl cellualse activity using soluble substrate sodium salt of CMC. You determine activity using a fixed amount of insoluble enzyme ( By weighing )> You may express your activity interms of protein also by liberating immobilized protein by chemical process (depending how you have immobilized )
As the native substrate, cellulose, is a water insoluble polymer, traditional reducing sugar assays using this substrate can not be employed for the measurement of cellulase activity. Analytical scientists have developed a number of alternative methods:
1) A viscometer can be used to measure the decrease in viscosity of a solution containing a water soluble cellulose derivative such as carboxymethyl cellulose upon incubation with a cellulase sample.The decrease in viscosity is directly proportional to the cellulase activity.
2) Cello-oligosaccharides can be chemically reduced through the action of sodium borohydride to produce their corresponding sugar alcohols. These compounds do not react in reducing sugar assays but their hydrolysis products do. This makes borohydride reduced cello-oligosaccharides valuable substrates for the assay of cellulase using traditional reducing sugar assays such as the Nelson-Symogyi method.
3) Enzymatic Assay of Cellulase using Sigma Aldrich spectrophotometric protocol.
- Badges
- Science topic
- Similar topics
- Papers