Int. J. Mol. Sci. 2011, 12(9), 5797-5814; doi:10.3390/ijms12095797
Article

Role of α-Helical Structure in Organic Solvent-Activated Homodimer of Elastase Strain K

1,* email, 1email, 2email and 1email
Received: 18 July 2011; in revised form: 5 August 2011 / Accepted: 11 August 2011 / Published: 9 September 2011
(This article belongs to the Special Issue Applications of Circular Dichroism)
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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
PDF Full-text Download PDF Full-Text [483 KB, Updated Version, uploaded 19 June 2014 03:58 CEST]
The original version is still available [483 KB, uploaded 19 June 2014 03:58 CEST]

Export to BibTeX |
EndNote


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.

AMA Style

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.

Chicago/Turabian Style

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.

Int. J. Mol. Sci. EISSN 1422-0067 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert