Purification and biochemical characteristic of a cold-active recombinant esterase from Pseudoalteromonas sp. 643A under denaturing conditions

• Autorzy: Dlugolecka A , Cieslinski H. , Bruzdziak P. , Gottfried K. , Turkiewicz M. , Kur J.
• Języki publikacji: EN

Abstrakty

EN

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°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 8M 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.

Słowa kluczowe

EN

purification; urea; denaturing condition; cold-active enzyme; enzyme; bacteria; Escherichia coli; lipolytic enzyme; esterase; Pseudoalteromonas; biochemical characteristics

• Wydawca: -
• Czasopismo: Polish Journal of Microbiology
• Rocznik: 2009
• Tom: 58
• Numer: 3
• Opis fizyczny: p.211-218,fig.,ref.

Twórcy

autor

Dlugolecka A

• Gdansk University of Technology, Narutowicza 11/12, 80-952 Gdansk, Poland

autor

Cieslinski H.

autor

Bruzdziak P.

autor

Gottfried K.

autor

Turkiewicz M.

autor

Kur J.

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