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PLoS One
2019 Apr 18;144:e0215396. doi: 10.1371/journal.pone.0215396.
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Characterization of novel hydrocarbon-degrading Gordonia paraffinivorans and Gordonia sihwensis strains isolated from composting.
Silva NM
,
de Oliveira AMSA
,
Pegorin S
,
Giusti CE
,
Ferrari VB
,
Barbosa D
,
Martins LF
,
Morais C
,
Setubal JC
,
Vasconcellos SP
,
da Silva AM
,
de Oliveira JCF
,
Pascon RC
,
Viana-Niero C
.
Abstract
Hydrocarbons are important environmental pollutants, and the isolation and characterization of new microorganisms with the ability to degrade these compounds are important for effective biodegradation. In this work we isolated and characterized several bacterial isolates from compost, a substrate rich in microbial diversity. The isolates were obtained from selective culture medium containing n-hexadecane, aiming to recover alkane-degraders. Six isolates identified as Gordonia by MALDI-TOF and 16S rRNA sequencing had the ability to degrade n-hexadecane in three days. Two isolates were selected for genomic and functional characterization, Gordonia paraffinivorans (MTZ052) and Gordonia sihwensis (MTZ096). The CG-MS results showed distinct n-hexadecane degradation rates for MTZ052 and MTZ096 (86% and 100% respectively). The genome sequence showed that MTZ052 encodes only one alkane degrading gene cluster, the CYP153 system, while MTZ096 harbors both the Alkane Hydroxylase (AH) and the CYP153 systems. qPCR showed that both gene clusters are induced by the presence of n-hexadecane in the growth medium, suggesting that G. paraffinivorans and G. sihwensis use these systems for degradation. Altogether, our results indicate that these Gordonia isolates have a good potential for biotransformation of hydrocarbons.
Fig 1. Phylogenetic analysis of MTZ052 and MTZ096.Maximum likelihood-based inference phylogenetic trees were reconstructed based on 57 gene families shared by the indicated bacterial species. Millisia brevis NBRC 105863 and Skermania piniformis NBRC 15059 were used as an outgroup. Branch bootstrap values are at least 80%, unless otherwise indicated.
Fig 2. Schematic comparison of the two main gene clusters for alkane degradation in MTZ052 and MTZ096.(A) Alkane hydroxylase cluster genes: alkane monooxygenase (red), rubredoxin (blue) and reductase (yellow). (B) CYP153 cluster genes: 2Fe-2S ferredoxin (red), cytochrome P450 (blue) and ferredoxin reductase (yellow). Transcriptional regulators colored in gray and pink are indicated. Comparison between the strains studied in this work (underlined) and available strains in IMG/M are shown.
Fig 3. Transcriptional pattern by real-time PCR.AH, (B) CYP153 gene clusters in G. sihwensis MTZ096 and (C) transcriptional pattern of CYP153 gene cluster in G. paraffinivorans MTZ052.
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