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. Author manuscript; available in PMC: 2015 Sep 21.
Published in final edited form as: J Mol Biol. 2015 Jan 12;427(6 0 0):1451–1463. doi: 10.1016/j.jmb.2015.01.003

Table 1.

Production and specific activity of FgCel5A and related variants

Enzyme1 Mutations Production (U/L)2 Specific activity (U/nmol)3
CBM CD 50°C 37°C
 FgCel5A - - 0.15 ± 0.06 1.19 ± 0.12 0.61 ± 0.06
 FgC5A.1 G10D T332I 0.80 ± 0.44 1.28 ± 0.07 0.66 ± 0.04
 FgC5A.2 G10D T166S, T332I, D342G 4.49 ± 1.24 1.25 ± 0.06 0.63 ± 0.03
 FgC5A.3 - T332I, D342G 4.51 ± 0.11 1.21 ± 0.09 0.65 ± 0.05
1

Each enzyme was expressed from pET28a without a Tat export signal or Bla reporter.

2

Cellulase production involved hydrolysis of CMC to a desired value and determination of the lysate volumes necessary to obtain such a value according to IUPAC standards. Hydrolysis was carried out using 1% CMC for 13 h at 37°C and sugar reducing ends were determined by reaction with DNS. Error is reported as the standard deviation from at least three biological replicates of the experiment.

3

Specific activity on CMC was measured using purified enzymes at 50°C and 37°C according to the IUPAC standard protocol for β-1,4-endoglucanase on CMC. Error is reported as the propagated error from three replicates of the experiment and error in the concentration measurement for the purified enzymes.