Novel Human Recombinant N-acetylgalactosamine-6-sulfate Sulfatase Produced in a Glyco-engineered Strain

Overview

Journal Heliyon

Specialty Social Sciences

Date 2024 Jul 2

PMID 38952373

Authors

Luisa N Pimentel-Vera

Alexander Rodriguez-Lopez

Angela J Espejo-Mojica

Aura Maria Ramirez

Carolina Cardona

Luis H Reyes

Shunji Tomatsu

Thapakorn Jaroentomeechai

Matthew P DeLisa

Oscar F Sanchez

Carlos J Almeciga-Diaz

Affiliations

Soon will be listed here.

Abstract

Mucopolysaccharidosis IVA (MPS IVA) is a lysosomal storage disease caused by mutations in the gene encoding the lysosomal enzyme -acetylgalactosamine-6-sulfate sulfatase (GALNS), resulting in the accumulation of keratan sulfate (KS) and chondroitin-6-sulfate (C6S). Previously, it was reported the production of an active human recombinant GALNS (rGALNS) in BL21(DE3). However, this recombinant enzyme was not taken up by HEK293 cells or MPS IVA skin fibroblasts. Here, we leveraged a glyco-engineered strain to produce a recombinant human GALNS bearing the eukaryotic trimannosyl core glycan, ManGlcNAc (rGALNSGly). The glycosylated GALNS was produced at 100 mL and 1.65 L scales, purified and characterized with respect to pH stability, enzyme kinetic parameters, cell uptake, and KS clearance. The results showed that the addition of trimannosyl core glycans enhanced both protein stability and substrate affinity. rGALNSGly was capture through a mannose receptor-mediated process. This enzyme was delivered to the lysosome, where it reduced KS storage in human MPS IVA fibroblasts. This study demonstrates the potential of a glyco-engineered for producing a fully functional GALNS enzyme. It may offer an economic approach for the biosynthesis of a therapeutic glycoprotein that could prove useful for MPS IVA treatment. This strategy could be extended to other lysosomal enzymes that rely on the presence of mannose N-glycans for cell uptake.

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