Draft Genome Sequence of Pseudomonas stutzeri Strain 19, an Isolate Capable of Efficient Degradation of Aromatic Hydrocarbons

ABSTRACT Pseudomonas stutzeri strain 19 is a Gram-negative bacterium capable of degrading aromatic hydrocarbons. The draft genome of P. stutzeri 19 is estimated to be 5.1 Mb, containing 4,652 protein-coding genes and a G+C content of 63.3%. Multiple genes responsible for the degradation of aromatics are present in this strain.

P seudomonas stutzeri strain 19 was isolated from a wastewater sample from Dayton, OH, USA. P. stutzeri 19 was shown, through gas chromatography-mass spectrometry (GC-MS) analysis, to efficiently metabolize toluene, xylenes, and 1,2,4-trimethyl benzene. Comparative BLAST analysis (http://blast.ncbi.nlm.nih.gov/Blast.cgi) of the 16S rRNA gene of P. stutzeri 19, identified using RNAmmer (1), showed 99% similarity with P. stutzeri DSM 4166 and P. stutzeri A1501, while Rapid Annotations using Subsystems Technology (RAST) identified P. stutzeri A1501 as the closest neighbor, with a score of 507. The genome of P. stutzeri was chosen for sequencing due to its ability to degrade recalcitrant aromatics and grow in harsh hydrocarbon-containing environments.
The NCBI PGAP predicted multiple genes involved in hydrocarbon degradation, including catechol 1,2-dioxygenase, homogentisate 1,2-dioxygenase, phenol monooxygenase, small and large subunits of benzoate 1,2-dioxygenase (benA and benB), alkane 1-monooxygenease, rubredoxin, alkene reductase, 2-alkenal reductase, P450, and a benzoate transporter protein, among others. BLAST analysis revealed two coding sequences with 99% homology to the alpha and beta subunits of toluene 1,2-dioxygenase of P. putida MT53 plasmid pWW53. Also, coding sequences with 96% homology to the xylene monooxygenase electron transfer subunit and 98% homology to the xylene monooxygenase hydrolase subunit of P. putida MT53 plasmid pWW53 were found. The presence of these enzymes explains the toluene and xylene degradation capacities of P. stutzeri 19. The genes for protocatechuate 3,4-dioxygenase, 3-carboxymuconate cycloisomerase, and 4-carboxymuconolactone decarboxylase of the central protocatechuate catabolic pathway for aromatic degradation were also present. A cluster of genes was observed with at least 78% homology to the ttg2 operon of P. putida that encodes an ABC transporter implicated in resistance to toluene (3,4). The genome of P. stutzeri 19 encodes many multidrug and heavy-metal resistance-nodulation-division (RND) efflux transporters, some of which have been associated with hydrocarbon resistance (5). The genome of P. stutzeri strain 19 will help to understand the adaptive mechanisms deployed by Gram-negative bacteria for survival and proliferation in hydrocarbons.
Accession number(s). This whole-genome shotgun project has been deposited at DDBJ/ENA/GenBank under the accession number NFZU00000000. The version described in this paper is NFZU01000000.

ACKNOWLEDGMENTS
This material is based on research sponsored by AFRL/RQTF under agreement FA8650-16-2-2605.
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