Screening results using different dilutions of CMBs.

The PesCDA^nm as well as an empty vector control were subjected to the fluorescamine-based screening. Using 3 glucosamine standards (15 μM, 50 μM, and 100 μM), the concentration of free primary amines were calculated. As expected, the empty plasmid control shows values close to 0, whereas PesCDA^nm shows an increasing amount of free primary amines formed with a decreasing dilution of CMBs. With a strong activity increase still being visible comparing the 1:10 to the 1:5 dilution, we settled on the 1:10 dilution assuming that the CMBs are saturated with enzymes. The results further show that with all dilutions less then 10% of the 500 μM A4 substrates were deacetylated. All values can be found in S1 Data.

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