Details

Autor(en) / Beteiligte

• Rodríguez, Ernesto
• Francia, Karen
• Brossard, Natalie
• García Vallejo, Juan J.
• Kalay, Hakan
• van Kooyk, Yvette
• Freire, Teresa
• Giacomini, Cecilia

Titel

Immobilization of β-galactosidase and α-mannosidase onto magnetic nanoparticles: A strategy for increasing the potentiality of valuable glycomic tools for glycosylation analysis and biological role determination of glycoconjugates

Ist Teil von

• Enzyme and microbial technology, 2018-10, Vol.117, p.45-55

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2018

Quelle

MEDLINE

Beschreibungen/Notizen

• [Display omitted] •A. oryzae β-galactosidase and C. ensiformis α-mannosidase were immobilized.•Immobilized glycosidases were useful for deglycosylation of model glycoproteins.•Immobilized glycosidases were re-used without loss of effectiveness.•Immobilized glycosidases were used for evaluation of biological role of glycans. Glycans present in biological glycoconjugates have several structural and functional roles. Elucidation of glycan structure and biological function is critical to understand their role in physiological and pathogenic process, enabling the development of diagnostic methods and disease treatment. Immobilized glycosidases are powerful tools for glycan analysis, as they are able to remove specific carbohydrates without altering the protein structure. Here we describe the individual immobilization of Aspergillus oryzae β-galactosidase and Canavalia ensiformis α-mannosidase onto agarose and silica magnetic nanoparticles activated with cyanate ester groups. High immobilization yields (70–90%) were achieved, keeping above 60% of its original activity. Immobilized glycosidases were effective in the selective deglycosylation of model glycoproteins and a Fasciola hepatica lysate, evidenced by a decrease in specific lectin recognition of 40–50% after enzymatic deglycosylation. Immobilized glycosidases were reused for several deglycosylation cycles without loss of effectiveness. Their use was extended to the elucidation of the glycan role of native glycoconjugates. A decrease in the recognition of lactoferrin treated with α-mannosidase by a C-type lectin receptor, DC-SIGN was found. Also the specific deglycosylation of a F. hepatica lysate demonstrated the relevance of mannosylated glycans in the induction of Th2/Treg immune responses during the infection. Our results show successful immobilization of specific glycosidases in nano-supports and validate their utility to identify glycans biological functions.