Details
| Original language | English |
|---|---|
| Pages (from-to) | 89-99 |
| Number of pages | 11 |
| Journal | Biochemical engineering journal |
| Volume | 147 |
| Early online date | 7 Apr 2019 |
| Publication status | Published - 15 Jul 2019 |
Abstract
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.
Keywords
- Bioactive peptides, Casein digestion, Enzyme immobilization, Membrane bioreactor, Subtilisin A, Surface functionalization
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biotechnology
- Environmental Science(all)
- Environmental Engineering
- Chemical Engineering(all)
- Bioengineering
- Engineering(all)
- Biomedical Engineering
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In: Biochemical engineering journal, Vol. 147, 15.07.2019, p. 89-99.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Proteolytic ceramic capillary membranes for the production of peptides under flow
AU - Antink, Marieke M.Hoog
AU - Sewczyk, Tim
AU - Kroll, Stephen
AU - Árki, Pál
AU - Beutel, Sascha
AU - Rezwan, Kurosch
AU - Maas, Michael
N1 - Funding Information: The authors are grateful for funding by the DFG through grant KR 3802/5-1 and to Novozymes A/S, Denmark, for providing the Alcalase® 2.5.
PY - 2019/7/15
Y1 - 2019/7/15
N2 - 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.
AB - 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.
KW - Bioactive peptides
KW - Casein digestion
KW - Enzyme immobilization
KW - Membrane bioreactor
KW - Subtilisin A
KW - Surface functionalization
UR - http://www.scopus.com/inward/record.url?scp=85064228826&partnerID=8YFLogxK
U2 - 10.1016/j.bej.2019.04.005
DO - 10.1016/j.bej.2019.04.005
M3 - Article
VL - 147
SP - 89
EP - 99
JO - Biochemical engineering journal
JF - Biochemical engineering journal
SN - 1369-703X
ER -