Table 1 Production and hydrolytic activity of WT and nine alanine-substituted mutants of TcCel7A.

From: Alanine substitution in cellobiohydrolase provides new insights into substrate threading

 

NC

WT

Entrance/

Base/

Loop/

W65A

S72A

N74A

N62A (Culture 1)

W63A

K203A

N125A

N222A

T223A

Production [mg/L]

<5

149

<5

171

229

<5

<5

117

168

126

213

Cellobiose

ND

0.778 (0.01)

ND

0.759 (0.016)

0.860 (0.026)

0.100 (0.013)

ND

0.768 (0.033)

0.882 (0.037)

0.603 (0.032)

0.703 (0.044)

Activity

ND

1.00 (0.01)

ND

0.851 (0.02)

0.720 (0.02)

>3.85 (0.49)

ND

1.26 (0.05)

1.01 (0.04)

0.908 (0.05)

0.634 (0.04)

  1. Production refers to the concentration (mg/L) of each enzyme obtained from the SDS-PAGE image in Fig. 3a. Cellobiose refers to the HPLC peak area of cellobiose (arbitrary unit) delivered by TcCel7A in the activity assay. Activity refers to hydrolytic activity, derived from dividing the Cellobiose value by the Production value. Average Cellobiose and Activity values are listed with standard deviations in parentheses. ND: Not detected. Higher activities are shown in bold (N62A and K203A).
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