Molecular detection and diversity of novel diterpenoid dioxygenase DitA1 genes from proteobacterial strains and soil samples

Resin acids are tricyclic diterpenoids naturally synthesized by trees that are released from wood during pulping processes. Using a newly designed primer set, genes similar to that encoding the DitA1 catalytic α-subunit of the diterpenoid dioxygenase, a key enzyme in abietane resin acid degradation...

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Published in:Environmental microbiology 2007-05, Vol.9 (5), p.1202-1218
Main Authors: Witzig, Robert, Aly, Hamdy A.H, Strömpl, Carsten, Wray, Victor, Junca, Howard, Pieper, Dietmar H
Format: Article
Language:English
Subjects:
Bacterial Proteins - classification
Bacterial Proteins - genetics
Burkholderia
Burkholderia - classification
Burkholderia - genetics
Burkholderia - metabolism
Dioxygenases - classification
Dioxygenases - genetics
Diterpenes - metabolism
Diterpenes, Abietane - metabolism
DNA Fingerprinting
Ferredoxins - classification
Ferredoxins - genetics
Molecular Sequence Data
Phylogeny
Proteobacteria - classification
Proteobacteria - genetics
Proteobacteria - physiology
Pseudomonas
Pseudomonas - classification
Pseudomonas - genetics
Pseudomonas - metabolism
Pseudomonas abietaniphila
Soil Microbiology
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Summary:Resin acids are tricyclic diterpenoids naturally synthesized by trees that are released from wood during pulping processes. Using a newly designed primer set, genes similar to that encoding the DitA1 catalytic α-subunit of the diterpenoid dioxygenase, a key enzyme in abietane resin acid degradation by Pseudomonas abietaniphila BKME-9, could be amplified from different Pseudomonas strains, whereas ditA1 gene sequence types representing distinct branches in the evolutionary tree were amplified from Burkholderia and Cupriavidus isolates. All isolates harbouring a ditA1-homologue were capable of growth on dehydroabietic acid as the sole source of carbon and energy and reverse transcription polymerase chain reaction analysis in three strains confirmed that ditA1 was expressed constitutively or in response to DhA, demonstrating its involvement in DhA-degradation. Evolutionary analyses indicate that gyrB (as a phylogenetic marker) and ditA1 genes have coevolved under purifying selection from their ancestral variants present in the most recent common ancestor of the genera Pseudomonas, Cupriavidus and Burkholderia. A polymerase chain reaction-single-strand conformation poylmorphism fingerprinting method was established to monitor the diversity of ditA1 genes in environmental samples. The molecular fingerprints indicated the presence ofa broad, previously unrecognized diversity of diterpenoid dioxygenase genes in soils, and suggest that other bacterial phyla may also harbour the genetic potential for DhA-degradation.
ISSN:1462-2912
1462-2920
DOI:10.1111/j.1462-2920.2007.01242.x