Molecular cloning, expression, and characterization of a novel endo-alpha-N-acetylgalactosaminidase from Enterococcus faecalis.
We report here the molecular cloning, expression and characterization of a novel endo-alpha-N-acetylgalactosaminidase, classified into the GH101 family, from Enterococcus faecalis (endo-EF). The recombinant endo-EF was found to catalyze the liberation of core1-disaccharides (Galbeta1-3GalNAc) from core1-pNP (Galbeta1-3GalNAcalpha-pNP) like other GH101 family enzymes. However, endo-EF seems to differ in specificity from the GH101 enzymes reported to date, because it was able to release trisaccharides from core2-pNP (Galbeta1-3[GlcNAcbeta1-6]GalNAcalpha-pNP) and tetrasaccharides from Gal-core2-pNP (Galbeta1-3[Galbeta1-3GlcNAcbeta1-6]GalNAcalpha-pNP). Interestingly, the enzyme could transfer not only core1-disaccharides but also core2-trisaccharides to alkanols generating alkyl-oligosaccharides. Endo-EF should facilitate O-glycoprotein research.
Goda
Hatsumi M
HM
Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan.
Ushigusa
Kota
K
Ito
Hiromi
H
Okino
Nozomu
N
Narimatsu
Hisashi
H
Ito
Makoto
M
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D002384:Catalysis; D003001:Cloning, Molecular; D004187:Disaccharides / Q000737:chemistry*; D013293:Enterococcus faecalis / Q000201:enzymology*; D006031:Glycosylation; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D013379:Substrate Specificity; D014312:Trisaccharides / Q000737:chemistry*; D048809:alpha-N-Acetylgalactosaminidase / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification
D001426:Bacterial Proteins; D004187:Disaccharides; D011994:Recombinant Proteins; D014312:Trisaccharides; D048809:alpha-N-Acetylgalactosaminidase