Alqueres_2011_Enzyme.Res_2011_316939

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 (TRX-PF2001Delta60) and without the thioredoxin tag (PF2001Delta60) were characterized, and various influences of Triton X-100 were determined. The optimal temperature for both enzymes was 80 degrees C. Although the thioredoxin presence did not influence the optimum temperature, the TRX-PF2001Delta60 presented specific activity twice lower than the enzyme PF2001Delta60. The enzyme PF2001Delta60 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 degrees C. However, the thermostability of the enzyme PF2001Delta60 was enhanced with Triton X-100.