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High-throughput screening for a moderately halophilic phenol-degrading strain and its salt tolerance response.

Lu ZY, Guo XJ, Li H, Huang ZZ, Lin KF, Liu YD - Int J Mol Sci (2015)

Bottom Line: Bacterial enrichments were cultivated in 48 deep well microplates instead of shake flasks or tubes.Measurement of phenol concentrations was performed in 96-well microplates instead of using the conventional spectrophotometric method or high-performance liquid chromatography (HPLC).PCR detection of the functional genes suggested that the largest subunit of multicomponent phenol hydroxylase (LmPH) and catechol 1,2-dioxygenase (C12O) were active in the phenol degradation process.

View Article: PubMed Central - PubMed

Affiliation: State Environmental Protection Key Laboratory of Environmental Risk Assessment and Control on Chemical Process, School of Resources and Environmental Engineering, East China University of Science and Technology, Shanghai 200237, China. lzy1009a@163.com.

ABSTRACT
A high-throughput screening system for moderately halophilic phenol-degrading bacteria from various habitats was developed to replace the conventional strain screening owing to its high efficiency. Bacterial enrichments were cultivated in 48 deep well microplates instead of shake flasks or tubes. Measurement of phenol concentrations was performed in 96-well microplates instead of using the conventional spectrophotometric method or high-performance liquid chromatography (HPLC). The high-throughput screening system was used to cultivate forty-three bacterial enrichments and gained a halophilic bacterial community E3 with the best phenol-degrading capability. Halomonas sp. strain 4-5 was isolated from the E3 community. Strain 4-5 was able to degrade more than 94% of the phenol (500 mg · L(-1) starting concentration) over a range of 3%-10% NaCl. Additionally, the strain accumulated the compatible solute, ectoine, with increasing salt concentrations. PCR detection of the functional genes suggested that the largest subunit of multicomponent phenol hydroxylase (LmPH) and catechol 1,2-dioxygenase (C12O) were active in the phenol degradation process.

No MeSH data available.


Related in: MedlinePlus

Phylogenetic tree based on 16S rRNA sequences, constructed by the neighbor-joining method, showing the position of strain 4-5 and representatives of some related strains. Pseudomonas putida NBRC 14671 was used as an out group. Bootstrap values were 1000 replicates, in which ≥50% were reported near the corresponding nodes. The scale bar indicates the percentage of genetic distance.
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ijms-16-11834-f005: Phylogenetic tree based on 16S rRNA sequences, constructed by the neighbor-joining method, showing the position of strain 4-5 and representatives of some related strains. Pseudomonas putida NBRC 14671 was used as an out group. Bootstrap values were 1000 replicates, in which ≥50% were reported near the corresponding nodes. The scale bar indicates the percentage of genetic distance.

Mentions: Strain 4-5 with optimal phenol-degrading characteristics was identified as a member of the Halomonas genus using 16S rDNA sequence analysis. Phylogenetic relationships between Halomonas sp. strain 4-5 and other species of the Halomonas genus were constructed on the basis of their 16S rRNA gene sequences (Figure 5). The closest relative of strain 4-5 was Halomonas xianhensis A-1 with a 16S rRNA sequence similarity of 99%. Phylogenetic analysis using the neighbor-joining algorithm with Pseudomonas putida NBRC 14671 as the out group also proved that the strain belonged to the Halomonas genus. The genus Halomonas was one of the largest among the moderately halophilic bacteria, which was originally proposed by Vreeland and primarily isolated from hypersaline or saline environments [18,19]. Members of the Halomonas genus were typically moderately halophilic bacteria, which contributed to the treatment of high-salinity industrial wastewater. H. xianhensis sp. nov. was first isolated by Zhao et al. [20] from the crude oil-contaminated soil in Shengli oilfield. In this study, the same species was isolated from the mineralized waste, and further studies of its salt tolerance response and degradation pathway were conducted.


High-throughput screening for a moderately halophilic phenol-degrading strain and its salt tolerance response.

Lu ZY, Guo XJ, Li H, Huang ZZ, Lin KF, Liu YD - Int J Mol Sci (2015)

Phylogenetic tree based on 16S rRNA sequences, constructed by the neighbor-joining method, showing the position of strain 4-5 and representatives of some related strains. Pseudomonas putida NBRC 14671 was used as an out group. Bootstrap values were 1000 replicates, in which ≥50% were reported near the corresponding nodes. The scale bar indicates the percentage of genetic distance.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4490417&req=5

ijms-16-11834-f005: Phylogenetic tree based on 16S rRNA sequences, constructed by the neighbor-joining method, showing the position of strain 4-5 and representatives of some related strains. Pseudomonas putida NBRC 14671 was used as an out group. Bootstrap values were 1000 replicates, in which ≥50% were reported near the corresponding nodes. The scale bar indicates the percentage of genetic distance.
Mentions: Strain 4-5 with optimal phenol-degrading characteristics was identified as a member of the Halomonas genus using 16S rDNA sequence analysis. Phylogenetic relationships between Halomonas sp. strain 4-5 and other species of the Halomonas genus were constructed on the basis of their 16S rRNA gene sequences (Figure 5). The closest relative of strain 4-5 was Halomonas xianhensis A-1 with a 16S rRNA sequence similarity of 99%. Phylogenetic analysis using the neighbor-joining algorithm with Pseudomonas putida NBRC 14671 as the out group also proved that the strain belonged to the Halomonas genus. The genus Halomonas was one of the largest among the moderately halophilic bacteria, which was originally proposed by Vreeland and primarily isolated from hypersaline or saline environments [18,19]. Members of the Halomonas genus were typically moderately halophilic bacteria, which contributed to the treatment of high-salinity industrial wastewater. H. xianhensis sp. nov. was first isolated by Zhao et al. [20] from the crude oil-contaminated soil in Shengli oilfield. In this study, the same species was isolated from the mineralized waste, and further studies of its salt tolerance response and degradation pathway were conducted.

Bottom Line: Bacterial enrichments were cultivated in 48 deep well microplates instead of shake flasks or tubes.Measurement of phenol concentrations was performed in 96-well microplates instead of using the conventional spectrophotometric method or high-performance liquid chromatography (HPLC).PCR detection of the functional genes suggested that the largest subunit of multicomponent phenol hydroxylase (LmPH) and catechol 1,2-dioxygenase (C12O) were active in the phenol degradation process.

View Article: PubMed Central - PubMed

Affiliation: State Environmental Protection Key Laboratory of Environmental Risk Assessment and Control on Chemical Process, School of Resources and Environmental Engineering, East China University of Science and Technology, Shanghai 200237, China. lzy1009a@163.com.

ABSTRACT
A high-throughput screening system for moderately halophilic phenol-degrading bacteria from various habitats was developed to replace the conventional strain screening owing to its high efficiency. Bacterial enrichments were cultivated in 48 deep well microplates instead of shake flasks or tubes. Measurement of phenol concentrations was performed in 96-well microplates instead of using the conventional spectrophotometric method or high-performance liquid chromatography (HPLC). The high-throughput screening system was used to cultivate forty-three bacterial enrichments and gained a halophilic bacterial community E3 with the best phenol-degrading capability. Halomonas sp. strain 4-5 was isolated from the E3 community. Strain 4-5 was able to degrade more than 94% of the phenol (500 mg · L(-1) starting concentration) over a range of 3%-10% NaCl. Additionally, the strain accumulated the compatible solute, ectoine, with increasing salt concentrations. PCR detection of the functional genes suggested that the largest subunit of multicomponent phenol hydroxylase (LmPH) and catechol 1,2-dioxygenase (C12O) were active in the phenol degradation process.

No MeSH data available.


Related in: MedlinePlus