Pseudomonas aeruginosa alkaline protease exhibits a high renaturation capability

  • Department of Immunobiology, Maria Curie-Sklodowska University, Lublin, Poland
  • Department of Immunobiology, Maria Curie-Sklodowska University, Lublin, Poland
  • Department of Genetics and Microbiology, Maria Curie-Skłodowska University, Lublin, Poland
  • Department of Cell Biology, Maria Curie-Skłodowska University, Lublin, Poland
  • Department of Cell Biology, Maria Curie-Skłodowska University, Lublin, Poland
  • Department of Cell Biology, Maria Curie-Skłodowska University, Lublin, Poland
  • Department of Immunobiology, Maria Curie-Sklodowska University, Lublin, Poland

Abstract

Thermally induced unfolding and renaturation capability of alkaline proteases (AprA) of three Pseudomonas aeruginosa strains, i.e. ATCC 27853 and two clinical isolates, was examined. Sequence analyses demonstrated a high level of aprA genes identity (99.24-99.8%) in these bacterial strains. The proteases retained 45-60% and 15% of their activity after pre-treatment at 60oC and 80oC, respectively, whereas pre-incubation at 90-95oC resulted in a higher level of activity than at 80oC. Zymography analyses and immunoblotting with AprA antiserum suggested a high thermostability and renaturation capability of the studied enzymes in comparison to another P. aeruginosa protease, elastase B. An intrinsic capability of renaturation of P. aeruginosa AprA was confirmed by fluorescence spectra of the native, thermally denatured, and renatured enzyme. The value of the fluorescence intensity of the denatured and subsequently cooled enzyme recovered to about 80% of the value of the native protein fluorescence intensity. Moreover, pre-incubation of the enzyme at 60oC and 90oC exerted only a slight effect on the intensity of absorbance and the shape of the amide I band, as demonstrated by Fourier transform infrared (FTIR) spectroscopy performed after subsequent cooling of the pre-treated enzyme. The results indicated a high renaturation capability of the P. aeruginosa AprA proteins.
Published
2019-03-04
Section
Articles