Enzymatic activity preservation and protection through entrapment within degradable hydrogels
Article first published online: 2 AUG 2013
© 2013 Wiley Periodicals, Inc.
Biotechnology and Bioengineering
Volume 110, Issue 11, pages 2994–3002, November 2013
How to Cite
Mariani, A. M., Natoli, M. E. and Kofinas, P. (2013), Enzymatic activity preservation and protection through entrapment within degradable hydrogels. Biotechnol. Bioeng., 110: 2994–3002. doi: 10.1002/bit.24971
- Issue published online: 21 SEP 2013
- Article first published online: 2 AUG 2013
- Accepted manuscript online: 7 JUN 2013 06:17AM EST
- Manuscript Accepted: 28 MAY 2013
- Manuscript Revised: 21 MAY 2013
- Manuscript Received: 2 JAN 2013
Additional supporting information may be found in the online version of this article at the publisher's web-site.
Figure S1. Lineweaver–Burke plots of PBS wash buffer removed from 30% PEGDA crosslinked gels where 1/[s] (1/M) is the concentration of ONPG substrate and 1/v (min*liter/mol) is 1/(the velocity of enzymatic conversion). a: 1st wash fluid at 3 h containing initial, non-entrapped enzyme, and unreacted reagents. b: 20 min later, 2nd wash fluid containing leached enzyme from the drop cast biogel. The apparent lack of linear regression fit proves no active enzyme leached into solution. c: 30% PEGDA crosslinked gels after 2nd wash proving maintained enzyme entrapment and no leaching at approximately 3 h post-entrapment.
Figure S2. Lineweaver–Burke plot of enzyme entrapped within 2% (w/v) alginate gels showing no linear regession fit and therefore no retention of activity. Where 1/[s] (1/M) is the concentration of ONPG substrate and 1/v (min*liter/mol) is 1/(the velocity of enzymatic conversion).
Figure S3. Sample graphs showing calculation methods for Vmax and Km. a: Conversion of ONPG substrate into ONP product at 3 h post-entrapment of various initial concentrations of substrate ONPG in 5% BIS crosslinked gel. b: Lineweaver–Burke plot of 5% BIS crosslinked gels at 3 h post-entrapment, where 1/[s] in 1/M is the concentration of substrate, and 1/v in min*liter/mol is 1/(the velocity of enzymatic conversion, seen as the slope in Supplementary Figure 3a).
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