I performed multi-enzyme immobilization on nanoparticles; after the enzyme immobilization on the support matrix, the supernatant was separated from the immobilized support. The supernatant contains enzymes (cellulase and xylanase) along with glutaraldehyde in a buffer solution of pH 4. The enzyme assays were performed for cellulase and xylanase activity in the supernatant using the DNS method, but no activity was detected in the supernatant. Anyhow, the ODs of the control tubes were much higher than the ODs of the test. What is the reason behind the increased OD of control than test in cellulase and xylanase activity? Does glutaraldehyde react directly with the DNS reagent, causing hindrance in enzyme activity and increased ODs of control?
you need to specify more clearly what your controls and test samples were.
The test of DNS assay contains an enzymatic mixture of cellulase and xylanase with glutaraldehyde. This mixture was incubated with the substrate in a water bath. Then, after incubation, the DNS assay was performed. In the control tube, the substrate was added, following the addition of a DNS reagent to block the binding sites of the substrate, and then the aforementioned enzyme mixture was added. The procedure of DNS assay was followed. Kindly answer @ Nick Chacos.
- Badges
- Science method