© 2006 Heron Publishing—Victoria, Canada
SSoNΔ and SSoNΔlong: two thermostable esterases from the same ORF in the archaeon Sulfolobus solfataricus?
Luigi Mandrich (1), Margherita Pezzullo (1), Mosè Rossi (1) and Giuseppe Manco (1, 2)
1. Institute of Protein Biochemistry, CNR, Via Pietro Castellino 111, 80131, Naples, Italy / 2. Corresponding author ([email protected]) / Received July 14, 2006; accepted October 13, 2006; published online November 20, 2006
Previously, we reported from the Sulfolobus solfataricus open reading frame (ORF) SSO2517 the cloning, overexpression and characterization of an esterase belonging to the hormone-sensitive lipase (HSL) family and apparently having a deletion at the N-terminus, which we named SSoNΔ. Searching the recently reported Sulfolobus acidocaldarius genome by sequence alignment, using SSO2517 as a query, allowed identity of a putative esterase (ORF SAC1105) sharing high sequence similarity (82%) with SSO2517. This esterase displays an N-terminus and total length similar to other known esterases of the HSL family. Analysis of the upstream DNA sequence of SS02517 revealed the possibility of expressing a longer version of the protein with an extended N-terminus; however, no clear translation signal consistent with a longer protein version was detected. This new version of SSO2517 was cloned, over-expressed, purified and characterized. The resulting protein, named SSoNΔlong, was 15-fold more active with the substrate p-nitrophenyl hexanoate than SSoNΔ. Furthermore, SSoNΔlong and SSoNΔ displayed different substrate specificities for triacylglycerols. These results and the phylogenetic relationship between S. solfataricus and S. acidocaldarius suggest a common origin of SSO2517 and SAC1105 from an ancestral gene, followed by divergent evolution. Alternatively, a yet-to-be discovered mechanism of translation that directs the expression of SSoNΔlong under specific metabolic conditions could be hypothesized.
Keywords: archaea, HSL family, N-terminus, thermophilic carboxylesterase.
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