TABLE 3.

Conversion of cellobiose, cellotetraose, xylobiose, lactose, and maltose determined in a Nimzyme assay

Cellobiose and cellotetraose were the only substrates that were hydrolyzed by JMB19063 among the substrates that we tested by Nimzyme assay. Each reaction was carried out in a 25-μl reaction volume containing 25 mm MES buffer (pH 6.5), 0.1 mm cellobiose or cellotetraose, and 2 μg (WT) or 10 μg (R587A or F598A) of recombinant enzyme at 50 °C for 15 min. All activities were corrected by the values of the negative control samples without enzyme. Data represents the means ± S.E. of three independent experiments.

	WT	R587A	F598A
Hydrolysis of cellobiose (%)	92.4 ± 2.4	4.8 ± 0.3	10.4 ± 1.2
Hydrolysis of cellotetraose (%)	61.9 ± 5.8	No activity	No activity