Figure 1.
Absorbance spectrum of cellulase:NS complex.
The spectrum of the cellulase:NS complex (black line) was captured after blanking the instrument with NS (red line) that had not been conjugated to cellulase enzymes. The clear peak at 280 nm and lack of additional peaks at absorbance wavelengths higher than 320 nm indicates that there was no interference in the readings from the nanospheres.
Figure 2.
Comparison of cellulase and cellulase:NS enzyme kinetics.
The velocity of glucose release is plotted as a function of A) insoluble cellulose concentration, and B) CMC concentration. Open circles, cellulase; black circles, cellulase-bead complex.
Table 1.
Kinetic parameters of glucose release from cellulose and CMC substrates using cellulase: NS complex compared to free cellulase enzyme.
Figure 3.
Efficiency of cellulose removal from wood cells by cellulase:NS.
A) A cellulose specific probe (CtCBM3-GFP) was used to fluorescently label cultured wood cells digested with either free cellulase (left) or cellulase:NS (right). Scale bar = 20 µm. B) The total fluorescence of individual wood cells was quantified and plotted as a function of cell area after degradation with cellulase (open circles) or cellulase-NS (closed circles). Inset shows the mean fluorescent intensity of each population of digested cells (n = 40), which are significantly different (*p<0.0001, Mann-Whitney test). Error bars = SD.
Figure 4.
Schematic diagram of enhanced glucose release from a crystalline cellulose substrate using the cellulase:NS construct relative to free cellulase.