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Squalene-hopene cyclase (Spterp25) from Streptomyces peucetius: sequence analysis, expression and functional
Gopal Prasad Ghimire1, Tae-Jin Oh, Hei Chan Lee
1Department of Pharmaceutical Engineering, Institute of Biomolecule Reconstruction (IBR), Sun Moon University, #100 Kalsan-ri, Tangjeong-myeon, Asansi, Chungnam 336-708, South Korea.
Abstract:
Squalene-hopene cyclase, which catalyzes the complex cyclization of squalene to the pentacyclic triterpene, hopene, is a key enzyme in the biosynthesis of hopanoids. The deduced amino acid sequence of the Streptomyces peucetius gene (spterp25) had significant similarity to other prokaryotic squalene-hopene cyclases. Like other triterpene cyclases, the S. peucetius squalene-hopene cyclase contains eight so-called QW-motifs with an aspartate-rich domain. The 2,025-bp squalene-hopene cyclase-encoding gene was expressed in Escherichia coli BL21(DE3)pLySs, and the in vitro activity of the recombinant cyclase was demonstrated using purified membrane protein. The cyclization product hopene was identified by gas chromatography/mass spectrometry (GC/MS).
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