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Enzyme Research
Volume 2011, Article ID 316939, 7 pages
Research Article

Characterization of the Recombinant Thermostable Lipase (Pf2001) from Pyrococcus furiosus: Effects of Thioredoxin Fusion Tag and Triton X-100

1Laboratório de Biologia Molecular, Programa de Biotecnologia e Biologia Molecular, IBqM, UFRJ, 21941-902 Rio de Janeiro, RJ, Brazil
2Laboratório de Biotecnologia Microbiana, Departamento de Bioquímica, IQ, UFRJ, 21941-909 Rio de Janeiro, RJ, Brazil
3Laboratório de Microbiologia Molecular e Proteínas, Departamento de Bioquímica, IQ, UFRJ, 21941-909 Rio de Janeiro, RJ, Brazil
4Laboratório de Bioprocessos, Programa de Engenharia Química, COPPE, UFRJ, 21945-970 Rio de Janeiro, RJ, Brazil

Received 14 January 2011; Revised 5 April 2011; Accepted 3 May 2011

Academic Editor: G. Viniegra-Gonzalez

Copyright © 2011 Sylvia Maria Campbell Alquéres et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


In this work, the lipase from Pyrococcus furiosus encoded by ORF PF2001 was expressed with a fusion protein (thioredoxin) in Escherichia coli. The purified enzymes with the thioredoxin tag (TRXPF2001Δ60) and without the thioredoxin tag (PF2001Δ60) were characterized, and various influences of Triton X-100 were determined. The optimal temperature for both enzymes was 80°C. Although the thioredoxin presence did not influence the optimum temperature, the TRXPF2001Δ60 presented specific activity twice lower than the enzyme PF2001Δ60. The enzyme PF2001Δ60 was assayed using MUF-acetate, MUF-heptanoate, and MUF-palmitate. MUF-heptanoate was the preferred substrate of this enzyme. The chelators EDTA and EGTA increased the enzyme activity by 97 and 70%, respectively. The surfactant Triton X-100 reduced the enzyme activity by 50% and lowered the optimum temperature to 60°C. However, the thermostability of the enzyme PF2001Δ60 was enhanced with Triton X-100.