A bacterial glycosidase enables mannose-6-phosphate modification and improved cellular uptake of yeast-produced recombinant human lysosomal enzymes

作者：Tiels Petra; Baranova Ekaterina; Piens Kathleen; De Visscher Charlotte; Pynaert Gwenda; Nerinckx Wim; Stout Jan; Fudalej Franck; Hulpiau Paco; Tannler Simon; Geysens Steven; Van Hecke Annelies; Valevska Albena; Vervecken Wouter; Remaut Han; Callewaert Nico^*

来源：Nature Biotechnology, 2012, 30(12): 1225-+.

DOI：10.1038/nbt.2427

Lysosomal storage diseases are treated with human lysosomal enzymes produced in mammalian cells. Such enzyme therapeutics contain relatively low levels of mannose-6-phosphate, which is required to target them to the lysosomes of patient cells. Here we describe a method for increasing mannose-6-phosphate modification of lysosomal enzymes produced in yeast. We identified a glycosidase from C. cellulans that %26apos;uncaps%26apos; N-glycans modified by yeast-type mannose-Pi-6-mannose to generate mammalian-type N-glycans with a mannose-6-phosphate substitution. Determination of the crystal structure of this glycosidase provided insight into its substrate specificity. We used this uncapping enzyme together with alpha-mannosidase to produce in yeast a form of the Pompe disease enzyme alpha-glucosidase rich in mannose-6-phosphate. Compared with the currently used therapeutic version, this form of alpha-glucosidase was more efficiently taken up by fibroblasts from Pompe disease patients, and it more effectively reduced cardiac muscular glycogen storage in a mouse model of the disease.

• 出版日期2012-12