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. 2015:2015:929424.
doi: 10.1155/2015/929424. Epub 2015 Aug 3.

Isolation and Characterization of Hydrocarbon-Degrading Yeast Strains from Petroleum Contaminated Industrial Wastewater

Boutheina Gargouri  1 , Najla Mhiri  1 , Fatma Karray  1 , Fathi Aloui  1 , Sami Sayadi  1
Affiliations

Isolation and Characterization of Hydrocarbon-Degrading Yeast Strains from Petroleum Contaminated Industrial Wastewater

Boutheina Gargouri et al. Biomed Res Int. 2015.

Abstract

Two yeast strains are enriched and isolated from industrial refinery wastewater. These strains were observed for their ability to utilize several classes of petroleum hydrocarbons substrates, such as n-alkanes and aromatic hydrocarbons as a sole carbon source. Phylogenetic analysis based on the D1/D2 variable domain and the ITS-region sequences indicated that strains HC1 and HC4 were members of the genera Candida and Trichosporon, respectively. The mechanism of hydrocarbon uptaking by yeast, Candida, and Trichosporon has been studied by means of the kinetic analysis of hydrocarbons-degrading yeasts growth and substrate assimilation. Biodegradation capacity and biomass quantity were daily measured during twelve days by gravimetric analysis and gas chromatography coupled with mass spectrometry techniques. Removal of n-alkanes indicated a strong ability of hydrocarbon biodegradation by the isolated yeast strains. These two strains grew on long-chain n-alkane, diesel oil, and crude oil but failed to grow on short-chain n-alkane and aromatic hydrocarbons. Growth measurement attributes of the isolates, using n-hexadecane, diesel oil, and crude oil as substrates, showed that strain HC1 had better degradation for hydrocarbon substrates than strain HC4. In conclusion, these yeast strains can be useful for the bioremediation process and decreasing petroleum pollution in wastewater contaminated with petroleum hydrocarbons.

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Figures

Figure 1
Figure 1
Biodegradation efficiency of yeast strains.
Figure 2
Figure 2
Phylogenetic dendogram data showing the taxonomic location of strains HC1 and HC4. (a) Neighbour-joining phylogenetic dendogram based on sequences of the D1/D2 domain of the 26S rRNA gene of HC1 and related taxa. Bootstrap values are given at the nodes. Scale bar represents the substitution percentage. Saccharomyces cerevisiae NRRL Y-12632T was used as outgroup. GenBank accession numbers follow species name in parenthesis. (b) Phylogenetic dendogram obtained by neighbour-joining analysis of the 5.8S-ITS sequence of HC4 and related taxa. Bootstrap values, determined from 100 replicates, are shown at branch nodes. Scale bar represents the substitution percentage. The outgroup we used was Tsuchiyaea wingfieldii CBS 7118T. GenBank accession numbers follow species name in parenthesis.
Figure 3
Figure 3
Hydrophobicity of yeast strains isolates during growth in mineral medium with 1.5% hexadecane or glucose as a sole carbon source. □ after growth on glucose; ■, after growth on hexadecane. Values are the mean of three separate experiments ± s.d.
Figure 4
Figure 4
Growth of selected yeasts on mineral medium with some alkanes (1% w/v) as the sole carbon source with respect to uninoculated controls over 12 days of degradation. (a) Candida tropicalis, (b) Trichosporon asahii.
Figure 5
Figure 5
Yeast cell count during 12 days. (a) Candida tropicalis, (b) Trichosporon asahii. Each point represents mean ± S.E of triple assays.
Figure 6
Figure 6
Gas chromatogram showing the biodegradation of alkane fraction of rich-hydrocarbon wastewater from Tunisian refinery by Candida tropicalis (a) and Trichosporon asahii (b) compared to the control.
Figure 7
Figure 7
Gas chromatogram showing the biotransformation of the aromatic fraction of rich-hydrocarbon wastewater from Tunisian refinery by Candida tropicalis (a) and Trichosporon asahii (b) compared to the control.
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