Limits...
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.

Show MeSH
PhzE gene fragment amplification of selected strains. Lane 1 and 10: 1 kb DNA-ladder; 2: positive control Pseudomonas chlororaphis subsp. chlororaphis DSM 50083T; 3: negative control (without template DNA); 4, 5, and 6: isolates H253, HB117, and LB129, respectively, exhibiting phzE fragments; 7, 8, and 9: isolates HB290, HB147, and LB164, respectively, exhibiting nor phzE fragments.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f3-marinedrugs-09-00772: PhzE gene fragment amplification of selected strains. Lane 1 and 10: 1 kb DNA-ladder; 2: positive control Pseudomonas chlororaphis subsp. chlororaphis DSM 50083T; 3: negative control (without template DNA); 4, 5, and 6: isolates H253, HB117, and LB129, respectively, exhibiting phzE fragments; 7, 8, and 9: isolates HB290, HB147, and LB164, respectively, exhibiting nor phzE fragments.

Mentions: Genes belonging to the phenazine biosynthetic pathway were present in approximately 10% of the bacterial strains analyzed. PCR results of 13 (8%) out of 164 bacterial strains and four reference organisms were positive in regard to the presence of phzE gene fragments (Figure 3, Tables 1 and 2). The investigated bacteria comprised different bacterial phyla, namely Actinobacteria (76), Bacteroidetes (2), Firmicutes (28) and Proteobacteria (62) (Table 1).


Genetic approach for the fast discovery of phenazine producing bacteria.

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

PhzE gene fragment amplification of selected strains. Lane 1 and 10: 1 kb DNA-ladder; 2: positive control Pseudomonas chlororaphis subsp. chlororaphis DSM 50083T; 3: negative control (without template DNA); 4, 5, and 6: isolates H253, HB117, and LB129, respectively, exhibiting phzE fragments; 7, 8, and 9: isolates HB290, HB147, and LB164, respectively, exhibiting nor phzE fragments.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f3-marinedrugs-09-00772: PhzE gene fragment amplification of selected strains. Lane 1 and 10: 1 kb DNA-ladder; 2: positive control Pseudomonas chlororaphis subsp. chlororaphis DSM 50083T; 3: negative control (without template DNA); 4, 5, and 6: isolates H253, HB117, and LB129, respectively, exhibiting phzE fragments; 7, 8, and 9: isolates HB290, HB147, and LB164, respectively, exhibiting nor phzE fragments.
Mentions: Genes belonging to the phenazine biosynthetic pathway were present in approximately 10% of the bacterial strains analyzed. PCR results of 13 (8%) out of 164 bacterial strains and four reference organisms were positive in regard to the presence of phzE gene fragments (Figure 3, Tables 1 and 2). The investigated bacteria comprised different bacterial phyla, namely Actinobacteria (76), Bacteroidetes (2), Firmicutes (28) and Proteobacteria (62) (Table 1).

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