TABLE 1.

Substrate specificities of 103S_EGCase I and M777_EGCase II

Name	Structure	Specific activitya		Reaction yieldb
		103S_EGCase I	M777_EGCase II	103S_EGCase I	M777_EGCase II
		nmol/min/μg		%	%
GlcCer	Glcβ1–1′Cer	<0.0001	<0.0001	0	0
GalCer	Galβ1–1′Cer	<0.0001	<0.0001	0	0
LacCer	Galβ1–4Glcβ1–1′Cer	1.80 ± 0.15	0.24 ± 0.02	62.8 ± 5.5	44.6 ± 5.1
GM3	NeuAcα2–3Galβ1–4Glcβ1–1′Cer	4.93 ± 0.64	0.61 ± 0.04	100 ± 3.0	95.9 ± 3.5
GM1	Galβ1–3GalNAcβ1–4 (NeuAcα2–3)Galβ1–4Glcβ1–1′Cer	6.44 ± 0.49	0.19 ± 0.03	100 ± 4.6	35.2 ± 4.3
Fucosyl-GM1	Fucα1–2Galβ1–3GalNAcβ1–4 (NeuAcα2–3)Galβ1–4Glcβ1–1′Cer	3.21 ± 0.25	<0.0001	65.8 ± 3.2	0
Gb4Cer	GalNAcβ1–3Galα1–4Galβ1–4Glcβ1–1′Cer	0.45 ± 0.05	<0.0001	95.4 ± 3.3	6.3 ± 1.5

^a The substrate specificity toward various GSLs is presented as the specific hydrolytic activities in the initial stage of the reaction. Values represent the means ±S.D. (n = 3).

^b The substrate specificity toward various GSLs is presented as the reaction yield (%) when the reaction reached equilibrium and was calculated using the equation, peak area for oligosacchrides ×100/peak area for 0.5 mm oligosaccharide. Values represent means ±S.D. (n = 3). Representative HPLC chromatograms of the reaction yield assay are presented in supplemental Fig. S2.