Table 1.

Thermal denaturation data and relative specific enzyme activity

GH 12 homolog	Variant	ΔTma	Tm (°C)b	Fit errorc	Activityd
T. reesei Ce112A	WT	0.0	54.4e	0.2	1.0
H. grisea Ce112A	WT	14.3	68.7e	0.3	0.2
T. reesei Ce112A	G170C	2.1	56.5e	0.2	1.2
	P201C	3.9	58.3e	0.2	0.8
	V210C	0.1	54.5e	0.1	1.8
	G170C/P201C	0.7	55.1e	0.1	0.4
	P201C/V210C	0.7	55.1	0.1	0.5
	G170C/V210C	ND	ND	ND	1.4
	G170C/P201C/V210C	0.0	54.5	0.3	0.1
H. grisea Ce112A	C175G	1.3	69.9	0.2	0.7
	C175S	0.2	68.9	0.1	0.7
	C206P	−9.1	59.5	0.2	0.9
	C206S	−5.4	63.3	0.1	1.5
	C216V	0.8	69.5	0.2	0.7
	C216S	−5.5	63.1	0.2	0.7

^a ΔT_m values are relative to T. reesei Ce112A WT, H. grisea Ce112A WT.

^b The thermal denaturation experiments were performed at 217 nm, in 0.05 M Bis-Tris propane, 0.05 M ammonium acetate (pH 8.0), and by increasing the temperature from 30°C to 90°C with data collected every 2°. The midpoint of the transition (T_m) is an apparent value because the thermal denaturation is not reversible.

^cT_m fit error corresponds to one standard deviation.

^d The specific enzyme activities was measured after 10 min incubation with oNPC, at 40°C, pH 5.5. The specific activities given are relative to T. reesei Ce112A WT, H. grisea Ce112A WT.

^e Two to five replicates were run with errors corresponding to one standard deviation between 0.15° and 0.42°.

ND, not determined.