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The biosynthetic genes for prenylated phenazines are located at two different chromosomal loci of Streptomyces cinnamonensis DSM 1042.

Seeger K, Flinspach K, Haug-Schifferdecker E, Kulik A, Gust B, Fiedler HP, Heide L - Microb Biotechnol (2010)

Bottom Line: The protein EpzP was expressed in Escherichia coli in form of a his-tag fusion protein and purified.The enzyme catalysed the prenylation of 5,10-dihydrophenazine-1-carboxylic acid (dihydro-PCA) using dimethylallyl diphosphate (DMAPP) as isoprenoid substrate.K(m) values were determined as 108 µM for dihydro-PCA and 25 µM for DMAPP.

View Article: PubMed Central - PubMed

Affiliation: Eberhard-Karls-University of Tübingen, Pharmaceutical Institute, Auf der Morgenstelle 8, D-72076 Tübingen, Germany.

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Related in: MedlinePlus

Gene cluster on cosmid 8‐4D of S. cinnamonensis DSM 1042, containing phenazine biosynthesis genes, mevalonate pathway genes and the prenyltransferase gene epzP. Further genes of endophenazine and furanonaphthoquinone I biosynthesis are contained in another genomic locus (Haagen et al., 2006).
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f2: Gene cluster on cosmid 8‐4D of S. cinnamonensis DSM 1042, containing phenazine biosynthesis genes, mevalonate pathway genes and the prenyltransferase gene epzP. Further genes of endophenazine and furanonaphthoquinone I biosynthesis are contained in another genomic locus (Haagen et al., 2006).

Mentions: Using conserved sequences of mevalonate pathway genes from different streptomycetes, we designed degenerate primers for the hydroxymethylglutaryl‐CoA synthase, hydroxymethylglutaryl‐CoA reductase and mevalonate diphosphate decarboxylase genes, using the codehop (COnsensus‐DEgenerate Hybrid Oligonucleotide Primer) program (Rose et al., 1998; 2003). A cosmid library of S. cinnamonensis had been established previously (Haagen et al., 2006) and was screened with the three primer pairs. This led to the identification of cosmid 8‐4D which gave PCR products for all three mevalonate pathway genes. This cosmid was subjected to full‐length sequencing using a shotgun library of DNA fragments. The cosmid insert comprised 45 167 bp and was deposited in the GenBank database under Accession No. HQ228364. Approximately in the middle of the insert sequence, we found the expected cluster of six mevalonate pathway genes, and directly upstream thereof a gene with obvious sequence similarity to the ABBA prenyltransferase class which was termed epzP (Fig. 2).


The biosynthetic genes for prenylated phenazines are located at two different chromosomal loci of Streptomyces cinnamonensis DSM 1042.

Seeger K, Flinspach K, Haug-Schifferdecker E, Kulik A, Gust B, Fiedler HP, Heide L - Microb Biotechnol (2010)

Gene cluster on cosmid 8‐4D of S. cinnamonensis DSM 1042, containing phenazine biosynthesis genes, mevalonate pathway genes and the prenyltransferase gene epzP. Further genes of endophenazine and furanonaphthoquinone I biosynthesis are contained in another genomic locus (Haagen et al., 2006).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3818865&req=5

f2: Gene cluster on cosmid 8‐4D of S. cinnamonensis DSM 1042, containing phenazine biosynthesis genes, mevalonate pathway genes and the prenyltransferase gene epzP. Further genes of endophenazine and furanonaphthoquinone I biosynthesis are contained in another genomic locus (Haagen et al., 2006).
Mentions: Using conserved sequences of mevalonate pathway genes from different streptomycetes, we designed degenerate primers for the hydroxymethylglutaryl‐CoA synthase, hydroxymethylglutaryl‐CoA reductase and mevalonate diphosphate decarboxylase genes, using the codehop (COnsensus‐DEgenerate Hybrid Oligonucleotide Primer) program (Rose et al., 1998; 2003). A cosmid library of S. cinnamonensis had been established previously (Haagen et al., 2006) and was screened with the three primer pairs. This led to the identification of cosmid 8‐4D which gave PCR products for all three mevalonate pathway genes. This cosmid was subjected to full‐length sequencing using a shotgun library of DNA fragments. The cosmid insert comprised 45 167 bp and was deposited in the GenBank database under Accession No. HQ228364. Approximately in the middle of the insert sequence, we found the expected cluster of six mevalonate pathway genes, and directly upstream thereof a gene with obvious sequence similarity to the ABBA prenyltransferase class which was termed epzP (Fig. 2).

Bottom Line: The protein EpzP was expressed in Escherichia coli in form of a his-tag fusion protein and purified.The enzyme catalysed the prenylation of 5,10-dihydrophenazine-1-carboxylic acid (dihydro-PCA) using dimethylallyl diphosphate (DMAPP) as isoprenoid substrate.K(m) values were determined as 108 µM for dihydro-PCA and 25 µM for DMAPP.

View Article: PubMed Central - PubMed

Affiliation: Eberhard-Karls-University of Tübingen, Pharmaceutical Institute, Auf der Morgenstelle 8, D-72076 Tübingen, Germany.

Show MeSH
Related in: MedlinePlus