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. 2022 Nov 25;10(12):2331.
doi: 10.3390/microorganisms10122331.

Comparative Proteomic Analysis of an Ethyl Tert-Butyl Ether-Degrading Bacterial Consortium

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Comparative Proteomic Analysis of an Ethyl Tert-Butyl Ether-Degrading Bacterial Consortium

Vijayalakshmi Gunasekaran et al. Microorganisms. .

Abstract

A bacterial consortium capable of degrading ethyl tert-butyl ether (ETBE) as a sole carbon source was enriched and isolated from gasoline-contaminated water. Arthrobacter sp., Herbaspirillum sp., Pseudacidovorax sp., Pseudomonas sp., and Xanthomonas sp. were identified as the initial populations with the 16S rDNA analysis. The consortium aerobically degraded 49% of 50 mg/L of ETBE, in 6 days. The ETBE degrading efficiency of the consortium increased to 98% even with the higher concentrations of ETBE (1000 mg/L) in the subsequent subcultures, which accumulated tert-butyl alcohol (TBA). Xanthomonas sp. and Pseudomonas sp. were identified as the predominant ETBE degrading populations in the final subculture. The metaproteome of the ETBE-grown bacterial consortium was compared with the glucose-grown bacterial consortium, using 2D-DIGE. Proteins related to the ETBE metabolism, stress response, carbon metabolism and chaperones were found to be abundant in the presence of ETBE while proteins related to cell division were less abundant. The metaproteomic study revealed that the ETBE does have an effect on the metabolism of the bacterial consortium. It also enabled us to understand the responses of the complex bacterial consortium to ETBE, thus revealing interesting facts about the ETBE degrading bacterial community.

Keywords: 2D-DIGE; ETBE; bacterial consortium; biodegradation; metaproteome.

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Conflict of interest statement

All the authors state that they have no conflict of interest.

Figures

Figure 1
Figure 1
Degradation of ETBE by bacterial consortium B (EB3). The graph represents the degradation assay carried out with consortium B (EB3) with 50 mg/L of ETBE as the initial concentration. formula image Control; formula image B consortium-EB3; formula image bacterial growth (OD at 600 nm). The error bars represent the standard deviation between the replicates (n = 2).
Figure 2
Figure 2
ETBE degradation and TBA formation in the degradation assay with different concentrations of ETBE by bacterial consortium B (EB11). The error bars represent the standard deviation between the replicates (n = 2). The graphs represent the degradation assay carried out with consortium B (EB11) with 50, 100, 500, and 1000 mg/L of ETBE as initial concentration.
Figure 3
Figure 3
2D-DIGE gel showing the electrophoretic map of the soluble proteins in bacterial consortium B. The image shows a representative picture of three independent gels. pI: Isoelectric point. Soluble proteins from bacterial consortium B grown in 0.5 g/L of glucose were labelled with Cy3 (green) and proteins from bacterial consortium B grown in 500 mg/L of ETBE were labelled with Cy5 (purple). The pooled internal standard was labelled with Cy2 (blue).
Figure 4
Figure 4
2D gel showing the selected protein spots. The image shows a preparative gel stained with Coomassie brilliant blue representing the 38 protein spots selected for analysis in MALDI-TOF/TOF. pI: Isoelectric point.
Figure 5
Figure 5
Schematic interpretation of the global response of bacterial consortium B in the presence of ETBE. Panel (A) represents the possible route of the ETBE metabolism followed in consortium B. Tert-butyl alcohol was identified by a GC-MS analysis, which is represented in the box. Panel (B) represents the proteins that are induced in bacterial consortium B in response to ETBE. The abundance of the protein is indicated in boxes with arrows pointing up or down to represent the more or less abundant proteins in the ETBE-induced state. Aldehyde dehydrogenase is identified in the proteomic analysis. It is believed to be involved in the ETBE metabolism and is abundant, as indicated by the arrow.

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