Journal
JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC
Volume 63, Issue 3-4, Pages 188-193Publisher
ELSEVIER
DOI: 10.1016/j.molcatb.2010.01.018
Keywords
Stem bromelain; Chemical modification; Pyromellitic dianhydride; Poly(maleic anhydride); Stability; Activity
Funding
- NSFC [50773009]
- Research Fund for the Doctoral Program of Higher Education [20090075120001]
- Sci & Tec Commission of Shanghai Municipality [08JC1400600]
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Chemical modification of lysine residues in stem bromelain was carried out using two novel reagents pyromellitic anhydride acid and poly(maleic anhydride). About 60% and 57% of the residues in bromelain were found to be modified by pyromellitic anhydride and poly(maleic anhydride), respectively. The modification brought about enhancement of thermal stability and the resistance to alkali and the surfactant. The optimum pH shifted from 7 to 9. Thermodynamic parameters, Delta H*, Delta G* and Delta S*, were determined as a function of temperature. The kinetic constants K-m of the modified enzymes were determined as 0.4092 x 10(-2) and 0.2825 x 10(-2) mol l(-1), respectively. SDS-PAGE profiling revealed a major bands of native and modified enzyme with molecular weights of 26 and 28.5 kDa. The results of FT-IR studies suggested that the modification caused only local structural changes. These results provide guidance for future development of stable protein formulations. Crown Copyright (C) 2010 Published by Elsevier B.V. All rights reserved.
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