| Full text | |
| Author(s): |
Liria, Cleber W.
[1]
;
Ungaro, Vitor A.
[1]
;
Fernandes, Raphaella M.
[1]
;
Costa, Natalia J. S.
[2]
;
Marana, Sandro R.
[1]
;
Rossi, Liane M.
[2]
;
Teresa Machini, M.
[1]
Total Authors: 7
|
| Affiliation: | [1] Univ Sao Paulo, Dept Biochem, Inst Chem, BR-05513970 Sao Paulo - Brazil
[2] Univ Sao Paulo, Dept Fundamental Chem, Inst Chem, BR-05513970 Sao Paulo - Brazil
Total Affiliations: 2
|
| Document type: | Journal article |
| Source: | JOURNAL OF NANOPARTICLE RESEARCH; v. 16, n. 11 NOV 2014. |
| Web of Science Citations: | 4 |
| Abstract | |
Enzyme-catalyzed chemical processes are selective, very productive, and generate little waste. Nevertheless, they may be optimized using enzymes bound to solid supports, which are particularly important for protease-mediated reactions since proteases undergo fast autolysis in solution. Magnetic nanoparticles are suitable supports for this purpose owing to their high specific surface area and to be easily separated from reaction media. Here we describe the immobilization of bovine alpha-chymotrypsin (alpha CT) on silica-coated superparamagnetic nanoparticles (Fe3O4@silica) and the characterization of the enzyme-nanoparticle hybrid (Fe3O4@silica-alpha CT) in terms of protein content, properties, recovery from reaction media, application, and reuse in enzyme-catalyzed peptide synthesis. The results revealed that (i) full acid hydrolysis of the immobilized protease followed by amino acid analysis of the hydrolyzate is a reliable method to determine immobilization yield; (ii) despite showing lower amidase activity and a lower K-cat/K-m value for a specific substrate than free alpha CT, the immobilized enzyme is chemically and thermally more stable, magnetically recoverable from reaction media, and can be consecutively reused for ten cycles to catalyze the amide bond hydrolysis and ester hydrolysis of the protected dipeptide Z-Ala-Phe-OMe. Altogether, these properties indicate the potential of Fe3O4@silica-alpha CT to act as an efficient, suitably stable, and reusable catalyst in amino acid, peptide, and protein chemistry as well as in proteomic studies. (AU) | |
| FAPESP's process: | 08/11695-1 - Alternative technologies in peptide chemistry: metal ion-promoted solvolysis, biocatalysis, conventional heating and microwave heating |
| Grantee: | Maria Teresa Machini |
| Support Opportunities: | Regular Research Grants |
| FAPESP's process: | 12/09068-4 - Microwaves, biocatalysis, divalent metal ions and nanotechnology in peptide synthesis |
| Grantee: | Maria Teresa Machini |
| Support Opportunities: | Regular Research Grants |