Novel GH109 enzymes for bioconversion of group A red blood cells to the universal donor group O

• Published in: New biotechnology 2023-11, Vol.77, p.130-138
• Main Authors: Curci, Nicola, Iacono, Roberta, Segura, Dorotea Raventos, Cillo, Michele, Cobucci-Ponzano, Beatrice, Strazzulli, Andrea, Leonardi, Antonio, Giger, Lars, Moracci, Marco
• Format: Article
• Language: English
• Subjects: Biocatalysis; Carbohydrate-active enzymes; Enzyme discovery; Glycoconjugates; Transfusion
• ISSN: 1871-6784; 1876-4347
• DOI: 10.1016/j.nbt.2023.08.002

Glycoside hydrolases (GHs) have been employed for industrial and biotechnological purposes and often play an important role in new applications. The red blood cell (RBC) antigen system depends on the composition of oligosaccharides on the surface of erythrocytes, thus defining the ABO blood type classification. Incorrect blood transfusions may lead to fatal consequences, making the availability of the correct blood group critical. In this regard, it has been demonstrated that some GHs may be helpful in the conversion of groups A and B blood types to produce group O universal donor blood. GHs belonging to the GH109 family are of particular interest for this application due to their ability to convert blood from group A to group O. This work describes the biochemical characterisation of three novel GH109 enzymes (NAg68, NAg69 and NAg71) and the exploration of their ability to produce enzymatically converted RBCs (ECO-RBC). The three enzymes showed superior specificity on pNP-α-N-acetylgalactosamine compared to previously reported GH109 enzymes. These novel enzymes were able to act on purified antigen-A trisaccharides and produce ECO-RBC from human donor blood. NAg71 converted type A RBC to group O with increased efficiency in the presence of dextran compared to a commercially available GH109, previously used for this application. •Characterisation of three highly efficient N-acetyl-α-GalNAcases from family GH109.•All these enzymes are able to hydrolyse erythrocyte A antigens trisaccharide.•Nag71 transform RBCs A into blood type O more efficiently than commercial NagA.