Proteolytic ceramic capillary membranes for the production of peptides under flow
Veröffentlichungsdatum
2019-04-15
Zusammenfassung
In this study, we investigate the effect of membrane surface functionalization on the immobilization of the protease subtilisin A and its performance in the production of peptides from the model protein casein under flow. The surface of tubular ceramic membranes was silanized to yield carboxylated and aminated supports for enzyme immobilization via non-covalent and carbodiimide activated binding. The protease density correlated with electrostatic interactions between the positively charged enzyme and the supports, with the highest enzyme density reached on negatively charged, carboxylated membranes (0.019 molecules/nm², noncovalent approach). Enzyme leaching was reduced by covalent binding of protease to carboxylated supports (5% leached) and slightly improved by binding to aminated membranes (46%) over non-covalent binding to unfunctionalized reference capillaries (66%). Regarding carbodiimide activated immobilization, protease on unfunctionalized and aminated supports exhibited a significantly larger specific activity (0.99 μmol/min/mg) than enzymes on carboxylated surfaces (0.15 μmol/min/mg), which suggests preferred enzyme orientation. In protein hydrolysis, these differences in surface-enzyme interactions were reflected by variations in peptide composition and degree of hydrolysis. Accordingly, we demonstrate that surface functionalization critically determines the surface properties of protease support materials for the production of peptides under flow and allows tailoring the performance of proteolytic capillary membranes.
Schlagwörter
Membrane bioreactor
;
Enzyme immobilization
;
SubtilisinA
;
Bioactive peptides
;
Casein digestion
;
Surface functionalization
;
Mimenima
Verlag
Elsevier
Institution
Institute
Dokumenttyp
Artikel/Aufsatz
Zeitschrift/Sammelwerk
Band
147
Startseite
89
Endseite
99
Zweitveröffentlichung
Ja
Dokumentversion
Postprint
Sprache
Englisch
Dateien![Vorschaubild]()
Lade...
Name
M. Hoog Antink,T. Sewczyk, S. Kroll, P. Árki, S. Beutel, K. Rezwan, M. Maas_Proteolytic ceramic capillary membranes for the production of peptides under flow_2019_accepted ver_Deckblatt_pdfa1.pdf
Size
2.89 MB
Format
Adobe PDF
Checksum
(MD5):4cffe85da3cc425b69a5455e3712f6ef