The biosynthetic genes for prenylated phenazines are located at two different chromosomal loci of Streptomyces cinnamonensis DSM 1042.
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.
Affiliation: Eberhard-Karls-University of Tübingen, Pharmaceutical Institute, Auf der Morgenstelle 8, D-72076 Tübingen, Germany.Show MeSH
Related in: MedlinePlus
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).
Affiliation: Eberhard-Karls-University of Tübingen, Pharmaceutical Institute, Auf der Morgenstelle 8, D-72076 Tübingen, Germany.