Cloning and characterization of the ddsA gene encoding decaprenyl diphosphate synthase from Rhodobacter capsulatus B10

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Abstract:

A decaprenyl diphosphate synthase gene (ddsA, GenBank accession No. DQ191802) was cloned from Rhodobacter capsulatus B10 by constructing and screening the genome library. An open reading frame of 1002 bp was revealed from sequence analysis. The deduced polypeptide consisted of 333 amino acids residues with an molecular mass of about 37 kDa. The DdsA protein contained the conserved amino acid sequence (DDXXD) of E-type polyprenyl diphosphate synthase and showed high similarity to others. In contrast, DdsA showed only 39% identity to a solanesyl diphosphate synthase cloned from R. capsulatus SB1003. DdsA was expressed successfully in Escherichia coli. Assaying the enzyme in vivo found it made E.coli synthesize UQ-10 in addition to the endogenous production UQ-8.Key words: ubiquinone, polyprenyl diphosphate synthase, gene expression, Rhodobacter capsulatus.

Un gène de décaprényle diphosphate synthase (ddsa, numéro d'acquisition GenBank DQ191802) fut cloné de Rhodobacter capsulatus B10 en construisant et criblant une librairie génomique. Un cadre de lecture ouvert de 1002 pb a pu être mis en évidence par analyse de la séquence. Le polypeptide déduit était formé de 333 résidus d'acides aminés ayant un poids moléculaire d'environ 37 kDa. La protéine DdsA contenait la séquence conservée en acides aminés (DDXXD) d'une polypényle diphosphate synthase de type E et a affiché une haute similitude aux autres. En revanche, DdsA n'a démontré que 39 % d'identité une solanésyle diphosphate synthase clonée de R. capsulatus SB1003. DdsA fut exprimé avec succès chez E. coli. Une analyse de l'enzyme in vivo a permis de constater qu'elle menait Escherichia coli à synthétiser du UQ-10 en plus de la production endogène de UQ-8.Mots clés : ubiquinone, polypényle diphosphate synthase, expression génique, Rhodobacter capsulatus.[Traduit par la Rédaction]

Document Type: Research Article

Publication date: December 1, 2006

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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