We grow the microbe in complex sugar media, spin it, take the supernatant, and add buffer and water. In the case of a test, we add the substrate and incubate it before adding DNSA, bringing it to a boil, adding Rocher's salt, and taking the reading. In the case of the control, we incubate the mixture without substrate, then DNSA is added, after which substrate is added, boiled, Rocher's salt is added, and reading is taken. The issue is that they both give the same reading. The control is Complex suger media with culture, buffer, water, and DNSA. Then, it is boiled, Rocher's salt is added, and a reading is taken. It is seen that less reading is seen, which suggests that the media is not the problem. I suspect the enzyme is intercellular. Can someone tell me any other possibility?
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