Izolovanje i delimično karakterisanje proteaze iz Pseudomonas aeruginosa ATCC 27853

Abstract

Enzymatic characteristics of a protease from a medically important, referent strain of Pseudomonas aeruginosa ATCC 27853 were determined. According to sodium dodecyl sulfate polyacrylamide gel electrophoresis, SDS-PAGE, and gel filtration, it was estimated that the molecular mass of the purified enzyme was about 15 kDa. Other enzymatic properties were found to be: pH optimum 7.1, pH stability between 6.5 and 10; temperature optimum around 60 °C while the enzyme was stable at 60°C for 30 min. Inhibition of the enzyme was observed with metal chelators, such as EDTA and 1,10-phenanthroline, suggesting that the protease is a metalloenzyme. Furthermore, the enzyme contains one mole of zinc ion per mole of enzyme. The protease was stable in the presence of different organic solvents, which enables its potential use for the synthesis of peptides.U ovom radu je okarakterisana ekstracelularna proteaza medicinski značajnog, referentnog soja Pseudomonas aeruginosa ATCC 27853. Molekulska masa prečišćenog enzima određena SDS PAGE i gel filtracijom iznosi oko 15 kDa. Određeni su sledeći enzimski parametri: pH optimum 7,1; pH stabilnost u opsegu 6,5-10; temperaturni optimum 60°C, a enzim je stabilan na 60°C 30 min. Na osnovu inhibicije enzima pomoću EDTA i 1,10-fenantrolina, utvrđeno je da proteaza predstavlja metaloenzim. Pokazano da proteaza sadrži 1 mol jona cinka po molu enzima. Proteaza je stabilna u prisustvu različitih organskih rastvarača, što omogućava upotrebu za sintezu peptida