dc.contributor.author |
Sharma, Rita |
|
dc.date.accessioned |
2021-09-27T07:57:29Z |
|
dc.date.available |
2021-09-27T07:57:29Z |
|
dc.date.issued |
2010-05-05 |
|
dc.identifier.uri |
https://www.sciencedirect.com/science/article/pii/S1359511310000334?via%3Dihub |
|
dc.identifier.uri |
http://dspace.bits-pilani.ac.in:8080/xmlui/handle/123456789/2196 |
|
dc.description.abstract |
Aminopeptidase from a solvent tolerant strain Pseudomonas aeruginosa PseA was purified and studied for its biochemical and molecular characteristics. Ion-exchange chromatography resulted in 11.9-fold purification and 38% recovery of the 56 kDa enzyme. The enzyme was found to be stable over a pH range of 6.0–8.0 and appreciably thermostable up to 70 °C. PseA aminopeptidase exhibited Km of 3.02 mM and Vmax of 6.71 μmol/mg/min towards l-Leu-p-nitroanilide. Remarkable stability in both hydrophilic and hydrophobic solvents makes PseA aminopeptidase unique. Partial N-terminal sequence of enzyme showed exact match with probable aminopeptidase of P. aeruginosa PAO1, coded by gene pepB. Polymerase chain reaction amplified the 1611-bp open reading frame encoding a 57.51 kDa, 536 amino acid PseA PepB polypeptide. The deduced PseA PepB protein sequence contained a 24-residue signal peptide (2.57 kDa) followed by a 1.28 kDa propeptide and a mature product of 500 residues. Search for conserved domain in PseA aminopeptidase explored its place in zinc-metallopeptidase family. Primary sequence analysis showed the hydrophobic inclination of the protein; and the 3D structure modeling elucidated the presence of a high content of hydrophobic residues on its surface probably imparting solvent stability to it. The enzyme might find potential applications in non-aqueous enzymology due to its marked thermostability and striking solvent stability. |
en_US |
dc.language.iso |
en |
en_US |
dc.publisher |
Elsiever |
en_US |
dc.subject |
Biology |
en_US |
dc.subject |
Pseudomonas aeruginosa |
en_US |
dc.subject |
Aminopeptidase |
en_US |
dc.subject |
Ion-exchange |
en_US |
dc.subject |
Chromatography |
en_US |
dc.title |
Purification and characterization of a solvent stable aminopeptidase from Pseudomonas aeruginosa: Cloning and analysis of aminopeptidase gene conferring solvent stability |
en_US |
dc.type |
Article |
en_US |