title: Identification of a GH110 subfamily of alpha 1,3-galactosidases: novel enzymes for removal of the alpha 3Gal xenotransplantation antigen.
abstract: In search of alpha-galactosidases with improved kinetic properties for removal of the immunodominant alpha1,3-linked galactose residues of blood group B antigens, we recently identified a novel prokaryotic family of alpha-galactosidases (CAZy GH110) with highly restricted substrate specificity and neutral pH optimum (Liu, Q. P., Sulzenbacher, G., Yuan, H., Bennett, E. P., Pietz, G., Saunders, K., Spence, J., Nudelman, E., Levery, S. B., White, T., Neveu, J. M., Lane, W. S., Bourne, Y., Olsson, M. L., Henrissat, B., and Clausen, H. (2007) Nat. Biotechnol. 25, 454-464). One member of this family from Bacteroides fragilis had exquisite substrate specificity for the branched blood group B structure Galalpha1-3(Fucalpha1-2)Gal, whereas linear oligosaccharides terminated by alpha1,3-linked galactose such as the immunodominant xenotransplantation epitope Galalpha1-3Galbeta1-4GlcNAc did not serve as substrates. Here we demonstrate the existence of two distinct subfamilies of GH110 in B. fragilis and thetaiotaomicron strains. Members of one subfamily have exclusive specificity for the branched blood group B structures, whereas members of a newly identified subfamily represent linkage specific alpha1,3-galactosidases that act equally well on both branched blood group B and linear alpha1,3Gal structures. We determined by one-dimensional (1)H NMR spectroscopy that GH110 enzymes function with an inverting mechanism, which is in striking contrast to all other known alpha-galactosidases that use a retaining mechanism. The novel GH110 subfamily offers enzymes with highly improved performance in enzymatic removal of the immunodominant alpha3Gal xenotransplantation epitope.
authors.affiliation: ZymeQuest Inc, Beverly, MA 01915, USA. pliu@zymequest.com
authors.forename: Qiyong P
authors.initials: QP
authors.lastname: Liu
authors.forename: Huaiping
authors.initials: H
authors.lastname: Yuan
authors.forename: Eric P
authors.initials: EP
authors.lastname: Bennett
authors.forename: Steven B
authors.initials: SB
authors.lastname: Levery
authors.forename: Edward
authors.initials: E
authors.lastname: Nudelman
authors.forename: Jean
authors.initials: J
authors.lastname: Spence
authors.forename: Greg
authors.initials: G
authors.lastname: Pietz
authors.forename: Kristen
authors.initials: K
authors.lastname: Saunders
authors.forename: Thayer
authors.initials: T
authors.lastname: White
authors.forename: Martin L
authors.initials: ML
authors.lastname: Olsson
authors.forename: Bernard
authors.initials: B
authors.lastname: Henrissat
authors.forename: Gerlind
authors.initials: G
authors.lastname: Sulzenbacher
authors.forename: Henrik
authors.initials: H
authors.lastname: Clausen
mesh_terms: D000818:Animals; D000941:Antigens / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D004912:Erythrocytes / Q000201:enzymology; D005434:Flow Cytometry; D005690:Galactose / Q000737:chemistry / Q000378:metabolism; D015870:Gene Expression; D006017:Glycolipids / Q000378:metabolism; D006868:Hydrolysis; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D010802:Phylogeny; D011817:Rabbits; D013237:Stereoisomerism; D013379:Substrate Specificity; D013552:Swine; D014183:Transplantation, Heterologous; D000519:alpha-Galactosidase / Q000145:classification / Q000235:genetics / Q000378:metabolism*
chemical_list: D000941:Antigens; D006017:Glycolipids; D000519:alpha-Galactosidase; D005690:Galactose