Preliminary crystal structure determination of the alkaline protease from the Antarctic psychrophile pseudomonas aeruginosa

Abstract

Abstract:A cold alkaline protease, isolated from an Antarctic Pseudomonas aeruginosa strain, has been purified and crystallized. Large crystals were obtained in the presence of PEG 6000 at pH 7 and pH 8. They belong to the space group P2,2,2,. A complete data set to 2.1 A resolution has been measured. The structure has been determined by the molecular replacement method using the coordinates of the mesophilic alkaline protease as a model. The molecular replacement solution displays a correlation coefficient of 0.39 and an R-factor of 0.48. Subsequent inspection of the electron density map in the active site region has confirmed the correctness of the solution. Model building and structure refinement will be initiated when the protease sequence becomes fully available. This is the second report, following one on an a-amylase, of the preliminary crystallographic characterization of a psychrophilic enzyme.

Comparative proteome analysis of psychrophilic versus mesophilic bacterial species: Insights into the molecular basis of cold adaptation of proteins

“…Many investigators have compared sequence and structure-based parameters among thermophilic and mesophilic proteins [ 10 ]. With the advent of pioneering efforts in late 1990's in solving three dimensional structures of cryophilic enzymes such as alpha-amylase [ 11 ]; alkaline protease [ 12 ]; triose phosphate isomerase [ 13 ]; malate dehydrogenase [ 14 ] from Antarctic microorganisms, and due to handful of available structures in the protein data bank (PDB), groups have focused to address the structural basis of proteins in cold adaptation [ 4 , 15 - 20 ].…”

Cellulases from psychrophilic microorganisms: a review

“…Therefore, 3D structural comparisons between mesophilic and psychrophilic cellulases are required in order to understand how proteins can adapt to low temperatures. The first report on the structure of a psychrophilic protein, a cold-active α-amylase from the antarctic psychrophile Alteromonas haloplanctis A23, came in the year 1996 [2], which was followed by a preliminary crystal structure determination of the alkaline protease from the antarctic psychrophile Pseudomonas aeruginosa [42]. However, the 3D structure of psychrophilic cellulases was not known till 2003, when a first paper appeared on the crystallization and preliminary X-ray crystallographic studies of a psychrophilic cellulase from Pseudoalteromonas haloplanktis [44].…”

Psychrozymes- The Next Generation Industrial Enzymes

“…Moreover use of cellulase can reduce the processing cost. Nilsson reported that cellulase enzymes are used to weaken the cellulose polymer and helps in dislodging the weakened fiber by mechanical process well known as "Biopolishing" [47].…”