Removal of N-linked glycans in cellobiohydrolase Cel7A from Trichoderma reesei reveals higher activity and binding affinity on crystalline cellulose

Table 1 Summary of TrCel7A WT and variants with the removed N-glycosylation motifs by site-directed mutagenesis

TrCel7A	Mutation	Expression host	T_m (°C)	Residual activity (%)
WT_Ao		Aspergillus oryzae	67.9	57 ± 1
N45Q_Ao	N45Q	Aspergillus oryzae	67.4	38 ± 2
N270Q_Ao	N270Q	Aspergillus oryzae	66.9	60 ± 3
N384Q_Ao	N384Q	Aspergillus oryzae	66.1	57 ± 3
ΔN-glyc_Ao	N45Q, N270Q, N384Q	Aspergillus oryzae	65.0	41 ± 2
WT_Tr		Trichoderma reesei	67.9	45 ± 2
ΔN-glyc_Tr	N45Q, N270Q, N384Q	Trichoderma reesei	65.2	44 ± 1

The mutated positions of N-glycosylation sites are presented in Fig. 2. The thermal stability, T_m, was determined from DSC. Residual activity is expressed as the activity after pre-incubation in the standard buffer for 1 h at T = T_m− 5 °C divided by the activity of the sample incubated at 25 °C. The activity was measured on 3 mM pNP-Lac in 50 mM sodium acetate buffer pH 5.0 for 30 min at 25 °C

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