title: Purification and biochemical characteristic of a cold-active recombinant esterase from Pseudoalteromonas sp. 643A under denaturing conditions.
abstract: In this paper production of a cold-active esterase EstA from the Antarctic bacterium Pseudoalteromonas sp. 643A in E. coli expression system was described. The purification and biochemical characteristic of EstA were performed in the presence of urea and then compared with results obtained for the esterase with no addition of urea and isolated from the native source. In both cases the cold-active enzyme displayed similar properties. However, the differences concerning thermal activity were observed. The optimal temperature for recombinant esterase in the presence of urea (1 M) was about 15 degrees C lower in comparison with enzyme isolated from the native source. Furthermore, the EstA was found to be more thermolabile in denaturant conditions. The differences were presumably caused by slightly changed protein structure in the presence of urea. The preservation of activity of EstA dissolved in buffer containing 8 M urea suggests that the protein structure is retained and it does not undergo dramatic changes due to high urea concentration. This thesis was confirmed with FT-IR data.
authors.affiliation: Department of Microbiology, Gdańsk University of Technology, Gdańsk, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Długołecka
authors.forename: Hubert
authors.initials: H
authors.lastname: Cieśliński
authors.forename: Piotr
authors.initials: P
authors.lastname: Bruździak
authors.forename: Karolina
authors.initials: K
authors.lastname: Gottfried
authors.forename: Marianna
authors.initials: M
authors.lastname: Turkiewicz
authors.forename: Józef
authors.initials: J
authors.lastname: Kur
mesh_terms: D003001:Cloning, Molecular; D003080:Cold Temperature; D004950:Esterases / Q000737:chemistry / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D006863:Hydrogen-Ion Concentration; D011489:Protein Denaturation; D031161:Pseudoalteromonas / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism
chemical_list: D004950:Esterases