Table 1.

Cofactor Requirements of PalH for the Hydrolysis of Chromogenic pNPαGlc^a

Additionsb	PalH Activityc
Control (PalH only)	NDAd
+ NAD+	0.92
+ Mn2+	NDA
+ DTT	NDA
+ EDTA	NDA
+ NAD+ + Mn2+	1.27
+ NAD+ + DTT	0.60
+ NAD+ + EDTA	NDA
+ NAD+ + Mn2+ + DTT	6.09
+ NAD+ + Mn2+ + DTT + EDTA	NDA

^aPalH used in these experiments had previously been dialyzed for 18 h against 25 mM Tris–HCl (pH 7.5) buffer.

^bAdditions to the basal 2-ml discontinuous assay were as described in Materials and Methods.

^cPalH activity expressed as μmol pNPαGlc hydrolyzed mg protein⁻¹ min⁻¹. Values are the mean of two separate assays.

^dNDA, No detectable PalH activity.