TABLE 1.

DNA cleavage rate, phage titer, and protein expression level analyses of SgrAI enzymes

SgrAI enzyme	Phage titera(PFU/μl)	DNA cleavage rate constant(min−1)
		Unactivatedb	Activatedc
WT	NPd	0.094 ± 0.15e	22 ± 7e
T4D	3.8 × 104 ± 0.5 × 104	0.058 ± 0.002	0.097 ± 0.009
S6D	4.3 × 104 ± 1.0 × 104	0.03 ± 0.01	0.19 ± 0.06
I7E	4.5 × 104 ± 0.3 × 104	0.091 ± 0.008	0.47 ± 0.09
R24E	3.7 × 104 ± 0.6 × 104	0.014 ± 0.003	0.028 ± 0.004
N25E	5.4 × 104 ± 0.7 × 104	0.05 ± 0.01	1.2 ± 0.2
P27W	6 × 104 ± 1 × 104	0.037 ± 0.05f	0.14 ± 0.01f
P27G	4.6 × 104 ± 0.4 × 104	0.06 ± 0.002f	0.12 ± 0.003f
Q34D	6.4 × 104 ± 1.2 × 104	0.04 ± 0.02	1.180 ± 0.2
I51E	4.9 × 104 ± 0.1 × 104	0.05 ± 0.01	0.26 ± 0.07
S56E	4.9 × 104 ± 0.3 × 104	0.08 ± 0.01g	0.17 ± 0.03g
S56Q	2.9 × 104 ± 0.3 × 104	0.08 ± 0.02g	5.5 ± 1.8g
A57E	4.6 × 104 ± 1.4 × 104	0.098 ± 0.002g	0.39 ± 0.06g
A57Q	NP	0.09 ± 0.01g	15 ± 8g
I59E	6.3 × 104 ± 0.2 × 104	0.14 ± 0.03	0.026 ± 0.006
M62E	2.6 × 104 ± 0.8 × 104	0.021 ± 0.001	3.2 ± 0.4
R84E	3.6 × 104 ± 0.5 × 104	0.082 ± 0.005	2.6 ± 0.2
R127A	6 × 104 ± 3 × 104	0.04 ± 0.04	7.6 ± 0.6
R131A	5.6 × 104 ± 0.5 × 104	0.10 ± 0.01g	0.28 ± 0.02g
R134A	5.2 × 104 ± 0.8 × 104	0.10 ± 0.013g	0.8 ± 0.2g

^aValue for the parent strain [Tuner (DE3) (Novagen, Inc.) with MspI.M] is 5.7 × 10⁴ ± 1.1 × 10⁴ PFU/μl.

^bSingle-turnover DNA cleavage rate constant of 1 nM ³²P-labeled 18-1 with 1 μM SgrAI enzyme.

^cSingle-turnover DNA cleavage rate constant of 1 nM ³²P-labeled 18-1 with 1 μM SgrAI enzyme and 1 μM PC DNA (activating DNA).

^dNP, no plaques detected.

^eFrom Park et al. (7).

^fFrom Park et al. (61).

^gFrom Shah et al. (16).