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Role of α-Helical Structure in Organic Solvent-Activated Homodimer of Elastase Strain K
Enzyme and Microbial Technology Laboratory, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, UPM Serdang 43400, Selangor, Malaysia
Enzyme and Microbial Technology Laboratory, Faculty of Science, Universiti Putra Malaysia, UPM Serdang 43400, Selangor, Malaysia
* Author to whom correspondence should be addressed.
Received: 18 July 2011; in revised form: 5 August 2011 / Accepted: 11 August 2011 / Published: 9 September 2011
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.
Keywords: P. aeruginosa strain K; organic solvent tolerant protease; dimerization; secondary structures
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MDPI and ACS Style
Abd. Rahman, R.N.Z.R.; Salleh, A.B.; Basri, M.; Wong, C.F. Role of α-Helical Structure in Organic Solvent-Activated Homodimer of Elastase Strain K. Int. J. Mol. Sci. 2011, 12, 5797-5814.
Abd. Rahman RNZR, Salleh AB, Basri M, Wong CF. Role of α-Helical Structure in Organic Solvent-Activated Homodimer of Elastase Strain K. International Journal of Molecular Sciences. 2011; 12(9):5797-5814.
Abd. Rahman, Raja Noor Zaliha Raja; Salleh, Abu Bakar; Basri, Mahiran; Wong, Chee Fah. 2011. "Role of α-Helical Structure in Organic Solvent-Activated Homodimer of Elastase Strain K." Int. J. Mol. Sci. 12, no. 9: 5797-5814.