Citation
Abstract
Recombinant elastase strain K overexpressed from E. coli KRX/pCon2(3) was purified to homogeneity by a combination of hydrophobic interaction chromatography and ion exchange chromatography, with a final yield of 48% and a 25-fold increase in specific activity. The purified protein had exhibited a first ever reported homodimer size of 65 kDa by SDS-PAGE and MALDI-TOF, a size which is totally distinct from that of typically reported 33 kDa monomer from P. aeruginosa. The organic solvent stability experiment had demonstrated a stability pattern which completely opposed the rules laid out in previous reports in which activity stability and enhancement were observed in hydrophilic organic solvents such as DMSO, methanol, ethanol and 1-propanol. The high stability and enhancement of the enzyme in hydrophilic solvents were explained from the view of alteration in secondary structures. Elastinolytic activation and stability were observed in 25 and 50% of methanol, respectively, despite slight reduction in α-helical structure caused upon the addition of the solvent. Further characterization experiments had postulated great stability and enhancement of elastase strain K in broad range of temperatures, pHs, metal ions, surfactants, denaturing agents and substrate specificity, indicating its potential application in detergent formulation.
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Official URL or Download Paper: http://www.mdpi.com/1422-0067/12/9/5797
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Additional Metadata
Item Type: | Article |
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Divisions: | Faculty of Biotechnology and Biomolecular Sciences Faculty of Science |
DOI Number: | https://doi.org/10.3390/ijms12095797 |
Publisher: | MDPI AG |
Keywords: | P. aeruginosa strain K; Organic solvent tolerant protease; Dimerization; Secondary structures |
Depositing User: | Nurul Ainie Mokhtar |
Date Deposited: | 01 Sep 2015 08:26 |
Last Modified: | 27 Sep 2016 01:27 |
Altmetrics: | http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.3390/ijms12095797 |
URI: | http://psasir.upm.edu.my/id/eprint/22407 |
Statistic Details: | View Download Statistic |
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