  Catalyzes the hydrolysis of the β-glycosidic linkage between oligosaccharides and ceramides in

     various glycosphingolipids.

  Oligosaccharide and ceramide remain intact, leaving them suitable for further analysis. 

  This is an Enzyme for Innovation (EFI). EFI is a project initiated by NEB to provide unique enzymes to

     the scientific community in the hopes of enabling the discovery of new and innovative applications.

    These enzymes have interesting properties and unique specificities.

Endoglycoceramidase I (EGCase I) catalyzes the hydrolysis of the β-glycosidic linkage between

oligosaccharides and ceramides in various glycosphingolipids. The enzyme leaves the oligosaccharide

and ceramide intact, keeping them suitable for further analysis. The enzyme efficiently releases

oligosaccharides from ganglio- and globo-type glycosphingolipids, certain type of cerebrosides

(glucosylcerebrosides), and exhibits low activity towards galactocerebrosides (See Reference 1,

FAQ #3). EGCase I does not hydrolyze sulfatides nor psychosine. 

EGCase I is isolated from a strain of E. coli, which contains the cloned EGCase I gene from Rhodococcus triatomea.

 1X EGCase I Reaction Buffer. Incubate at 37°C

10X EGCase I Reaction Buffer

1X EGCase I Reaction Buffer 
0.1% Triton® X-100 
50 mM sodium acetate 
(pH 5.2 @ 25°C)

Apparent: 50 kDa

One unit of R. triatomea EGCase I was defined as the amount of enzyme required to hydrolyze 1 μmol of

ganglioside GM1a per minute at 37°C.

Unit Assay Conditions
Two fold dilutions of EGCase I are incubated with 10 nmoles of GM1a substrate in 1X EGCase I Reaction

Buffer in a 10 µl reaction. The reaction mix is incubated for 5 minutes at 37°C. Released glycans are

labeled with 2-AB, and reaction products are analyzed by ultra-performance hydrophilic interaction liquid

chromatography with fluorescence detection.

65°C for 20 min.

 1. Albrecht, S. et al. (2016). Analytical Chemistry. 88, 4795-4802.