TABLE 1.

Substrate specificity of recombinant NgaP

pNP-glycosides (50 nmol) were incubated with the enzyme at 37 °C for 30 min in 100 μl of 25 mm sodium acetate buffer (pH 6.0). Values are the means of triplicate determinations. ND, not determined. 10 nmol of each GSL was incubated at 37 °C for 24 h with 10 milliunits of the enzyme in 20 μl of 25 mm sodium acetate (pH 6.0). Values are the means of duplicate determinations. GalCer, galactosylceramide.

pNP-substrate	Hydrolysis
	1 milliunit of NgaP	3 milliunits of NgaP	30 milliunits of NgaP
	%
pNP-β-GalNAc	55.2	100	100
pNP-β-GlcNAc	0	0.851	2.84
pNP-β-Gal	ND	ND	0
pNP-β-Glc	ND	ND	0
pNP-β-Fuc	ND	ND	0
pNP-β-Xyl	ND	ND	0
pNP-α-GalNAc	ND	ND	0
pNP-α-GlcNAc	ND	ND	0
pNP-α-Gal	ND	ND	0
pNP-α-Glc	ND	ND	0
pNP-α-Fuc	ND	ND	0
pNP-α-Xyl	ND	ND	0

GSL	Structure	Relative activity
		%
GA2	GalNAcβ1–4Galβ1–4Glcβ1–1′Cer	100
GM2	GalNAcβ1–4(NeuAcα2–3)Galβ1–4Glcβ1–1′Cer	0
GA1	Galβ1–3GalNAcβ1–4Galβ1–4Glcβ1–1′Cer	0
GM1a	Galβ1–3GalNAcβ1–4(NeuAcα2–3)Galβ1–4Glcβ1–1′Cer	0
Gb4Cer	GalNAcβ1–3Galα1–4Galβ1–4Glcβ1–1′Cer	39.6
Gb3Cer	Galα1–4Galβ1–4Glcβ1–1′Cer	0
LacCer	Galβ1–4Glcβ1–1′Cer	0
GlcCer	Glcβ1–1′Cer	0
GalCer	Galβ1–1′Cer	0
Sulfatide	HSO3-3Galβ1–1′Cer	0