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Identification and Characterization of a New Type III Polyketide Synthase from a Marine Yeast,Naganishia uzbekistanensis
Authors:Laure Martinelli  Vanessa Redou  Bastien Cochereau  Ludovic Delage  Nolwenn Hymery  Elisabeth Poirier  Christophe Le Meur  Gaetan Le Foch  Lionel Cladiere  Mohamed Mehiri  Nathalie Demont-Caulet  Laurence Meslet-Cladiere
Abstract:A putative Type III Polyketide synthase (PKSIII) encoding gene was identified from a marine yeast, Naganishia uzbekistanensis strain Mo29 (UBOCC-A-208024) (formerly named as Cryptococcus sp.) isolated from deep-sea hydrothermal vents. This gene is part of a distinct phylogenetic branch compared to all known terrestrial fungal sequences. This new gene encodes a C-terminus extension of 74 amino acids compared to other known PKSIII proteins like Neurospora crassa. Full-length and reduced versions of this PKSIII were successfully cloned and overexpressed in a bacterial host, Escherichia coli BL21 (DE3). Both proteins showed the same activity, suggesting that additional amino acid residues at the C-terminus are probably not required for biochemical functions. We demonstrated by LC-ESI-MS/MS that these two recombinant PKSIII proteins could only produce tri- and tetraketide pyrones and alkylresorcinols using only long fatty acid chain from C8 to C16 acyl-CoAs as starter units, in presence of malonyl-CoA. In addition, we showed that some of these molecules exhibit cytotoxic activities against several cancer cell lines.
Keywords:marine yeast  PKSIII  triketide pyrones  pentaketide resorcinols  cytotoxic activity
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