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Fig. 3 | Journal of Biological Engineering

Fig. 3

From: Rational protein design of Bacillus sp. MN chitosanase for altered substrate binding and production of specific chitosan oligomers

Fig. 3

Kinetic parameters of CSN, CSN-Y270A, CSN-E59A, and CSN-W118A on different substrates. The catalytic rate constant kcat at 30 °C is illustrated for the substrates (a) chitosan polymer DA 1.5%, (b) D4, and (c) D5. Also, (d) a direct comparison between chitosan polymer DA 1.5% and D4 is given. Three independent enzyme batches were used for each enzyme, and the kinetics were performed as triplicates for each batch. Data given are the mean values of all nine replicates, and the standard deviations between the three independent enzyme batches are indicated. A 30 min incubation period was used for CSN-E59A for D4 and for CSN-W118A. All other reactions were incubated for 10 min. Different enzyme concentrations were used for chitosan polymer DA 1.5% (0.05 μM CSN, 0.3 μM CSN-Y270A, 0.05 μM CSN-E59A, and 2.5 μM CSN-W118A), D4 (0.04 μM CSN, 0.023 μM CSN-Y270A, and 0.1 μM CSN-E59A), and D5 (0.02 μM CSN, 0.168 μM CSN-Y270A, 0.2 μM CSN-E59A, and 1 μM CSN-W118A)

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