Table 3. ΔT _m and kinetic parameters of wild-type BglA and mutant enzymes.

No.	Mutants	ΔT m (°C)	k cat (sec−1)	K m (mM)	k cat/K m (sec−1 mM−1)
0	Wild-type	0.0a	44.73±0.81	1.21±0.03	36.89±0.38
1	E96Kb	3.7	28.42±0.15	0.19±0.01	149.00±4.97
	E96Rb	4.8	21.19±0.50	0.47±0.02	44.81±1.08
	E96D	−1	n.d.	n.d.	n.d.
2	Q141Kb	−0.3	n.d.	n.d.	n.d.
3	Q216K	–0.5	30.62±0.54	0.47±0.04	64.90±4.90
	Q216R	–3.5	26.02±0.12	0.46±0.00	56.87±0.19
	Q216D	0.1	n.d.	n.d.	n.d.
	Q216E	−2.2	n.d.	n.d.	n.d.
4	Q277K	0.4	n.d.	n.d.	n.d.
	Q277R	0.0	n.d.	n.d.	n.d.
	Q277Db	−0.1	1.04±0.02	18.06±0.29	17.41±0.12
	Q277Eb	0.7	0.85±0.31	15.68±3.50	19.00±2.72
5	T231Kb	1.2	36.32±1.36	0.61±0.07	59.31±4.96
	T231R	−0.3	n.d.	n.d.	n.d.
	T231D	−3.8	n.d.	n.d.	n.d.
	T231E	−3.2	n.d.	n.d.	n.d.
	T231D/D388K	4.8	32.76±0.76	0.19±0.01	174.90±9.48
6	N437Kb	8.8	34.85±0.18	0.50±0.01	69.74±1.45
	N437R	5.3	10.27±0.51	0.40±0.06	25.57±2.39
	N437D	−0.4	n.d.	n.d.	n.d.
	N437E	−0.4	n.d.	n.d.	n.d.
7	S440Kb	1.3	1.63±0.03	24.39±0.30	14.99±0.14
	S440R	2.4	3.96±0.32	31.78±1.90	8.04±0.17
	S440D	−0.7	n.d.	n.d.	n.d.
	S440E	0.5	n.d.	n.d.	n.d.
8	E96R/T231D/D388K/N437K	15.7	10.27±0.51	0.40±0.06	25.57±2.39

^a T _m of wild-type protein was detected to be 38.5°C.

^bSuggested mutation.

n.d.: Not determined.