title: Bacterial glycosidases for the production of universal red blood cells.
abstract: Enzymatic removal of blood group ABO antigens to develop universal red blood cells (RBCs) was a pioneering vision originally proposed more than 25 years ago. Although the feasibility of this approach was demonstrated in clinical trials for group B RBCs, a major obstacle in translating this technology to clinical practice has been the lack of efficient glycosidase enzymes. Here we report two bacterial glycosidase gene families that provide enzymes capable of efficient removal of A and B antigens at neutral pH with low consumption of recombinant enzymes. The crystal structure of a member of the alpha-N-acetylgalactosaminidase family reveals an unusual catalytic mechanism involving NAD+. The enzymatic conversion processes we describe hold promise for achieving the goal of producing universal RBCs, which would improve the blood supply while enhancing the safety of clinical transfusions.
authors.affiliation: ZymeQuest Inc., 100 Cummings Center, Suite 436H, Beverly, Massachusetts 01915, USA.
authors.forename: Qiyong P
authors.initials: QP
authors.lastname: Liu
authors.forename: Gerlind
authors.initials: G
authors.lastname: Sulzenbacher
authors.forename: Huaiping
authors.initials: H
authors.lastname: Yuan
authors.forename: Eric P
authors.initials: EP
authors.lastname: Bennett
authors.forename: Greg
authors.initials: G
authors.lastname: Pietz
authors.forename: Kristen
authors.initials: K
authors.lastname: Saunders
authors.forename: Jean
authors.initials: J
authors.lastname: Spence
authors.forename: Edward
authors.initials: E
authors.lastname: Nudelman
authors.forename: Steven B
authors.initials: SB
authors.lastname: Levery
authors.forename: Thayer
authors.initials: T
authors.lastname: White
authors.forename: John M
authors.initials: JM
authors.lastname: Neveu
authors.forename: William S
authors.initials: WS
authors.lastname: Lane
authors.forename: Yves
authors.initials: Y
authors.lastname: Bourne
authors.forename: Martin L
authors.initials: ML
authors.lastname: Olsson
authors.forename: Bernard
authors.initials: B
authors.lastname: Henrissat
authors.forename: Henrik
authors.initials: H
authors.lastname: Clausen
mesh_terms: D000017:ABO Blood-Group System / Q000737:chemistry; D001419:Bacteria / Q000201:enzymology*; D001665:Binding Sites; D001788:Blood Grouping and Crossmatching; D002384:Catalysis; D002855:Chromatography, Thin Layer; D004912:Erythrocytes / Q000378:metabolism*; D005434:Flow Cytometry; D006026:Glycoside Hydrolases / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D011387:Prokaryotic Cells / Q000201:enzymology; D017433:Protein Structure, Secondary; D013379:Substrate Specificity; D017186:Titrimetry; D048809:alpha-N-Acetylgalactosaminidase / Q000737:chemistry
chemical_list: D000017:ABO Blood-Group System; D006026:Glycoside Hydrolases; D048809:alpha-N-Acetylgalactosaminidase