A thermo-stable lysine aminopeptidase from Pseudomonas aeruginosa: Isolation, purification, characterization, and sequence analysis

Journal of Basic Microbiology
Yan Tao WuYa Ping Tian

Abstract

Pseudomonas aeruginosa NJ-814, isolated from garden soil, produced an extracellular aminopeptidase that was purified using ammonium sulfate precipitation and ion exchange chromatography. The purity was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the Mr value of the enzyme was estimated to be 55 kDa. The purified enzyme shows maximum activity at pH 9.0 and 80 °C. It exhibits high thermo-stability. Half of the activity can remain after incubation at 80 °C for 119 min. It is stable within pH range of 7.5-10.5. It is strongly activated by Co(2+) and inhibited by Fe(2+) , Cu(2+) , Ni(2+) , Zn(2+) , and ethylene diamine tetraacetic acid (EDTA). The specificity of the enzyme was investigated. Within several aminoacyl-p-nitroanilines (AA-pNA), Lys-pNA is proven to be the optimal substrate. The Michaelis-Menten constant (Km ) of the enzyme for Lys-pNA and Leu-pNA were 2.32 and 9.41 mM, respectively. Peptide map fingerprinting shows that the sequence of the enzyme is highly similar to aminopeptidase Y from P. aeruginosa 18A. It can be speculated that this enzyme is a Zn(2+) -dependent enzyme and contains two zinc ions in its active site.

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Citations

Sep 16, 2016·World Journal of Microbiology & Biotechnology·Hongyu YangYaping Tian
Jan 11, 2018·APL Bioengineering·Federica RigoldiAlfonso Gautieri
Apr 29, 2020·Applied Microbiology and Biotechnology·Arya Nandan, K M Nampoothiri

Related Concepts

Aminopeptidase
Enzyme Stability
Hydrogen-Ion Concentration
Metals
Pseudomonas aeruginosa
Soil Microbiology
Substrate Specificity
Ammonium Sulfate
Physiologic Thermoregulation
Ion-Exchange Chromatography Procedure

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