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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

An integrated procedure for high-throughput screening. (a) High-throughput screening for the degrading strain; (b) Traditional method for the degrading strain.
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ijms-16-11834-f001: An integrated procedure for high-throughput screening. (a) High-throughput screening for the degrading strain; (b) Traditional method for the degrading strain.

Mentions: The halophilic bacterial communities were cultivated in 48 deep well microplates under different NaCl concentrations. The high-throughput screening procedure (Figure 1) was designed based on two characteristics, salt tolerance and phenol degradation, for environmental samples. Using this process, we screened halophilic bacterial communities from 43 bacterial enrichments and harvested a bacterial community that could degrade phenol efficiently in a broad range of salinities. Previous studies have reported halophilic or halotolerant microorganisms from salt environments using conventional screening techniques. For example, Leitão et al. [14] isolated a halotolerant strain of Penicillium chrysogenum from a salt mine in Portugal that could degrade at least 300 mg·L−1 phenol. Arulazhagan et al. [15] enriched a halotolerant bacterial consortium from mixed saline water samples collected from India. Haddadi and Shavandi [16] isolated a moderately halophilic strain of Halomonas sp. strain PH2-2 from petroleum-contaminated soil in Iran that could degrade 400 mg·L−1 phenol with a removal efficiency of 95% at 7% NaCl. However, these conventional screenings were tedious, had a large material requirement and low-throughput, which were applied to limited samples. This high-throughput screening system processed 43 samples for phenol degradation experiments targeting six salinity levels, and only six microplates were needed to replace 258 flasks or tubes, which greatly reduced the volume occupied by the shaker and the reagent consumption. Compared with shake flasks or tubes handling samples one by one, deep well microplates were intensive in cultivating samples simultaneously. Therefore, the screening was condensed and performed with small sample quantities. It could be used for the mass cultivation of phenol-degrading enrichments from various saline habitats.


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)

An integrated procedure for high-throughput screening. (a) High-throughput screening for the degrading strain; (b) Traditional method for the degrading strain.
© Copyright Policy
Related In: Results  -  Collection

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

ijms-16-11834-f001: An integrated procedure for high-throughput screening. (a) High-throughput screening for the degrading strain; (b) Traditional method for the degrading strain.
Mentions: The halophilic bacterial communities were cultivated in 48 deep well microplates under different NaCl concentrations. The high-throughput screening procedure (Figure 1) was designed based on two characteristics, salt tolerance and phenol degradation, for environmental samples. Using this process, we screened halophilic bacterial communities from 43 bacterial enrichments and harvested a bacterial community that could degrade phenol efficiently in a broad range of salinities. Previous studies have reported halophilic or halotolerant microorganisms from salt environments using conventional screening techniques. For example, Leitão et al. [14] isolated a halotolerant strain of Penicillium chrysogenum from a salt mine in Portugal that could degrade at least 300 mg·L−1 phenol. Arulazhagan et al. [15] enriched a halotolerant bacterial consortium from mixed saline water samples collected from India. Haddadi and Shavandi [16] isolated a moderately halophilic strain of Halomonas sp. strain PH2-2 from petroleum-contaminated soil in Iran that could degrade 400 mg·L−1 phenol with a removal efficiency of 95% at 7% NaCl. However, these conventional screenings were tedious, had a large material requirement and low-throughput, which were applied to limited samples. This high-throughput screening system processed 43 samples for phenol degradation experiments targeting six salinity levels, and only six microplates were needed to replace 258 flasks or tubes, which greatly reduced the volume occupied by the shaker and the reagent consumption. Compared with shake flasks or tubes handling samples one by one, deep well microplates were intensive in cultivating samples simultaneously. Therefore, the screening was condensed and performed with small sample quantities. It could be used for the mass cultivation of phenol-degrading enrichments from various saline habitats.

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