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Genetic approach for the fast discovery of phenazine producing bacteria.

Schneemann I, Wiese J, Kunz AL, Imhoff JF - Mar Drugs (2011)

Bottom Line: The genetic potential for phenazine production was shown for four type strains belonging to the genera Streptomyces and Pseudomonas as well as for 13 environmental isolates from marine habitats.Phenazine production was demonstrated for the type strains known to produce endophenazines, 2-hydroxy-phenazine, phenazine-1-carboxylic acid, phenazine-1,6-dicarboxylic acid, and chlororaphin as well as for members of marine Actinobacteria.Interestingly, a number of unidentified phenazines possibly represent new phenazine structures.

View Article: PubMed Central - PubMed

Affiliation: Kieler Wirkstoff-Zentrum (KiWiZ) am Leibniz-Institut für Meereswissenschaften (IFM-GEOMAR), Am Kiel-Kanal 44, 24106 Kiel, Germany.

ABSTRACT
A fast and efficient approach was established to identify bacteria possessing the potential to biosynthesize phenazines, which are of special interest regarding their antimicrobial activities. Sequences of phzE genes, which are part of the phenazine biosynthetic pathway, were used to design one universal primer system and to analyze the ability of bacteria to produce phenazine. Diverse bacteria from different marine habitats and belonging to six major phylogenetic lines were investigated. Bacteria exhibiting phzE gene fragments affiliated to Firmicutes, Alpha- and Gammaproteobacteria, and Actinobacteria. Thus, these are the first primers for amplifying gene fragments from Firmicutes and Alphaproteobacteria. The genetic potential for phenazine production was shown for four type strains belonging to the genera Streptomyces and Pseudomonas as well as for 13 environmental isolates from marine habitats. For the first time, the genetic ability of phenazine biosynthesis was verified by analyzing the metabolite pattern of all PCR-positive strains via HPLC-UV/MS. Phenazine production was demonstrated for the type strains known to produce endophenazines, 2-hydroxy-phenazine, phenazine-1-carboxylic acid, phenazine-1,6-dicarboxylic acid, and chlororaphin as well as for members of marine Actinobacteria. Interestingly, a number of unidentified phenazines possibly represent new phenazine structures.

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Schematic representation of the phenazine biosynthesis pathway.
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f1-marinedrugs-09-00772: Schematic representation of the phenazine biosynthesis pathway.

Mentions: Although principal parts of the phenazine biosynthesis (Figure 1) and their genetics are known [15,16], there are still gaps in our knowledge and the phenazine pathway is still hypothetical [17]. The function and gene products of phzC, phzE and phzD have been experimentally proven. These enzymes catalyze the conversion into 3-deoxy-d-arabino-heptulosonic acid 7-phosphate (DAHP), 2-amino-2-deoxyisochorismic acid (ADIC) and trans-2,3-dihydro-3-hydroxyanthranilic acid (DHHA) [17–20]. The phenazine core structure is synthesized subsequently by condensation of two molecules of 6-amino-5-oxocyclohex-2-ene-1-carboxylic acid [17]. Presumably, PhzF is responsible for the isomerization of DHHA [18]. By now, the function of the enzymes PhzA, PhzB and PhzG are almost enlightened [19] and their role in the condensation and rearrangement reactions to form PCA is demonstrated [17]. According to this information, the design of appropriate oligonucleotide probes and primers was possible. Mavrodi et al. (2010) [21] quite recently designed four different pairs of oligonucleotides on the basis of the phzF gene of the phenazine biosynthesis and investigated the diversity of phenazine producers mainly in soil-dwelling and plant-associated bacteria.


Genetic approach for the fast discovery of phenazine producing bacteria.

Schneemann I, Wiese J, Kunz AL, Imhoff JF - Mar Drugs (2011)

Schematic representation of the phenazine biosynthesis pathway.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3111181&req=5

f1-marinedrugs-09-00772: Schematic representation of the phenazine biosynthesis pathway.
Mentions: Although principal parts of the phenazine biosynthesis (Figure 1) and their genetics are known [15,16], there are still gaps in our knowledge and the phenazine pathway is still hypothetical [17]. The function and gene products of phzC, phzE and phzD have been experimentally proven. These enzymes catalyze the conversion into 3-deoxy-d-arabino-heptulosonic acid 7-phosphate (DAHP), 2-amino-2-deoxyisochorismic acid (ADIC) and trans-2,3-dihydro-3-hydroxyanthranilic acid (DHHA) [17–20]. The phenazine core structure is synthesized subsequently by condensation of two molecules of 6-amino-5-oxocyclohex-2-ene-1-carboxylic acid [17]. Presumably, PhzF is responsible for the isomerization of DHHA [18]. By now, the function of the enzymes PhzA, PhzB and PhzG are almost enlightened [19] and their role in the condensation and rearrangement reactions to form PCA is demonstrated [17]. According to this information, the design of appropriate oligonucleotide probes and primers was possible. Mavrodi et al. (2010) [21] quite recently designed four different pairs of oligonucleotides on the basis of the phzF gene of the phenazine biosynthesis and investigated the diversity of phenazine producers mainly in soil-dwelling and plant-associated bacteria.

Bottom Line: The genetic potential for phenazine production was shown for four type strains belonging to the genera Streptomyces and Pseudomonas as well as for 13 environmental isolates from marine habitats.Phenazine production was demonstrated for the type strains known to produce endophenazines, 2-hydroxy-phenazine, phenazine-1-carboxylic acid, phenazine-1,6-dicarboxylic acid, and chlororaphin as well as for members of marine Actinobacteria.Interestingly, a number of unidentified phenazines possibly represent new phenazine structures.

View Article: PubMed Central - PubMed

Affiliation: Kieler Wirkstoff-Zentrum (KiWiZ) am Leibniz-Institut für Meereswissenschaften (IFM-GEOMAR), Am Kiel-Kanal 44, 24106 Kiel, Germany.

ABSTRACT
A fast and efficient approach was established to identify bacteria possessing the potential to biosynthesize phenazines, which are of special interest regarding their antimicrobial activities. Sequences of phzE genes, which are part of the phenazine biosynthetic pathway, were used to design one universal primer system and to analyze the ability of bacteria to produce phenazine. Diverse bacteria from different marine habitats and belonging to six major phylogenetic lines were investigated. Bacteria exhibiting phzE gene fragments affiliated to Firmicutes, Alpha- and Gammaproteobacteria, and Actinobacteria. Thus, these are the first primers for amplifying gene fragments from Firmicutes and Alphaproteobacteria. The genetic potential for phenazine production was shown for four type strains belonging to the genera Streptomyces and Pseudomonas as well as for 13 environmental isolates from marine habitats. For the first time, the genetic ability of phenazine biosynthesis was verified by analyzing the metabolite pattern of all PCR-positive strains via HPLC-UV/MS. Phenazine production was demonstrated for the type strains known to produce endophenazines, 2-hydroxy-phenazine, phenazine-1-carboxylic acid, phenazine-1,6-dicarboxylic acid, and chlororaphin as well as for members of marine Actinobacteria. Interestingly, a number of unidentified phenazines possibly represent new phenazine structures.

Show MeSH