Improving the Catalytic Activity of Hyperthermophilic Pyrococcus horikoshii Prolidase for Detoxification of Organophosphorus Nerve Agents over a Broad Range of Temperatures

Table 1

Substrate specificity of recombinant wild type and variant P. horikoshii prolidases with different proline dipeptides and a single-proline tripeptide.

Substrate	Relative Activity (%) of WT-Ph1prol-specific activity
	WT-; Ph1prol	A195T/; G306S-; Ph1prol	Y301C/; K342N-; Ph1prol	E127G/; E252D-; Ph1prol	E36V-; Ph1prol
Val-Pro	100	33	36	46	56
	(4,602)
Met-Pro	100	143	33	117	77
	(2,809)
Phe-Pro	100	116	53	53	68
	(2,199)
Leu-Pro	100	116	36	90	65
	(2,132)
Ala-Pro	100	219	183	324	556
	(1,452)
Gly-Pro	100	94	52	107	444
	(369)
Pro-Ala	100	137	33	75	332
	(14)
Val-Leu-Pro	100	0	88	0	0
	(2)

Prolidase assays were performed at 100°C and contained 0.2 mM CoCl₂ and 4 mM of each substrate. One hundred percent specific activity is reported for WT-Ph1prol and correlates to U/mg in parentheses below the 100% relative activity.