Role of α-helical structure in organic solvent-activated homodimer of elastase strain K
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 fi...
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MDPI AG
2011
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| Online Access: | http://psasir.upm.edu.my/id/eprint/22407/ http://psasir.upm.edu.my/id/eprint/22407/1/Role%20of%20%CE%B1-Helical%20Structure%20in%20Organic%20Solvent-Activated%20Homodimer%20of%20Elastase%20Strain%20K.pdf |
| _version_ | 1848844476091465728 |
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| author | Raja Abdul Rahman, Raja Noor Zaliha Salleh, Abu Bakar Basri, Mahiran Wong, Chee Fah |
| author_facet | Raja Abdul Rahman, Raja Noor Zaliha Salleh, Abu Bakar Basri, Mahiran Wong, Chee Fah |
| author_sort | Raja Abdul Rahman, Raja Noor Zaliha |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | 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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| format | Article |
| id | upm-22407 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T08:31:31Z |
| publishDate | 2011 |
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| spelling | upm-224072016-09-27T01:27:29Z http://psasir.upm.edu.my/id/eprint/22407/ Role of α-helical structure in organic solvent-activated homodimer of elastase strain K Raja Abdul Rahman, Raja Noor Zaliha Salleh, Abu Bakar Basri, Mahiran Wong, Chee Fah 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. MDPI AG 2011 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/22407/1/Role%20of%20%CE%B1-Helical%20Structure%20in%20Organic%20Solvent-Activated%20Homodimer%20of%20Elastase%20Strain%20K.pdf Raja Abdul Rahman, Raja Noor Zaliha and Salleh, Abu Bakar and Basri, Mahiran and Wong, Chee Fah (2011) Role of α-helical structure in organic solvent-activated homodimer of elastase strain K. International Journal of Molecular Sciences, 12 (9). pp. 5797-5814. ISSN 1422-0067 http://www.mdpi.com/1422-0067/12/9/5797 10.3390/ijms12095797 |
| spellingShingle | Raja Abdul Rahman, Raja Noor Zaliha Salleh, Abu Bakar Basri, Mahiran Wong, Chee Fah Role of α-helical structure in organic solvent-activated homodimer of elastase strain K |
| title | Role of α-helical structure in organic solvent-activated homodimer of elastase strain K |
| title_full | Role of α-helical structure in organic solvent-activated homodimer of elastase strain K |
| title_fullStr | Role of α-helical structure in organic solvent-activated homodimer of elastase strain K |
| title_full_unstemmed | Role of α-helical structure in organic solvent-activated homodimer of elastase strain K |
| title_short | Role of α-helical structure in organic solvent-activated homodimer of elastase strain K |
| title_sort | role of α-helical structure in organic solvent-activated homodimer of elastase strain k |
| url | http://psasir.upm.edu.my/id/eprint/22407/ http://psasir.upm.edu.my/id/eprint/22407/ http://psasir.upm.edu.my/id/eprint/22407/ http://psasir.upm.edu.my/id/eprint/22407/1/Role%20of%20%CE%B1-Helical%20Structure%20in%20Organic%20Solvent-Activated%20Homodimer%20of%20Elastase%20Strain%20K.pdf |