Optimization of Potential Phenol Degrading Pseudomonas Strain KP-3 Isolated from Soil Sample and Characterization by Using Gen III Microlog (original) (raw)
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Desalination, 2009
Using the phenol enrichment cultivation method, we succeeded in screening a few bacterial strains with a varied ability for utilizing phenol as the sole carbon source from pharmaceutical disposal wastewaters plant. Among these strains, a potent phenol-degrading bacterium, assigned Pseudomonas sp. SA01, was identified and examined as having the greatest potential for degrading phenol. According to biochemical characteristics and 16S rRNA sequence analysis, the isolate was identified as Pseudomonas sp. The isolated strain started to degrade 0.7 g/l of phenol after an initial very short lag phase, and phenol decomposition was then rapidly completed within 30 h. Pseudomonas sp. SA01 was able to degrade phenol in concentrations up to 1 g/l. Higher phenol concentrations (>1 g/l) had a significant inhibitory effect on bacterial growth. The optimum degradation pH value was found to be 6.5. Addition of mannitol and casein as auxiliary carbon and nitrogen sources enhanced the rate of phenol removal to as low as 20 h. Based on the absorption spectra of catechol bioconversion of phenol-grown cells, it was concluded that the SA01 strain metabolizes the phenol via a meta-cleavage pathway.
Phenol is a toxic pollutant found in effluent of numerous industries and its elimination is a foremost challenge. The utilization of bacteria plays a crucial role in phenol bioremediation. For isolation of phenol degrading bacteria, sample was collected from industrial waste and enriched in mineral salt medium (MSM) contained 300 mg/L phenol. The strain was identified based on 16S rRNA gene analysis as Pseudomonas species and the phylogenetic analysis affiliated the strain with Pseudomonas monteilii(AF064458) as the most closely related species. Phenol tolerance of the strain in MSM supplemented with various concentrations of phenol indicates that the strain NCCP-407 can grow best at 750 mg L−1 phenol. The strain showed complete degradation of 750 mg L−1 phenol in 56 hours when supplement as a sole source of carbon and energy with the average degradation rate of 28 mg L−1h−1. The doubling time was recorded approximately as 12.49 h−1. The present study suggests that this strain is efficient in phenol degradation and can be used in treatment of wastewater containing phenol.
Molecules
Phenols are very soluble in water; as a result, they can pollute a massive volume of fresh water, wastewater, groundwater, oceans, and soil, negatively affecting plant germination and animal and human health. For the detoxification and bioremediation of phenol in wastewater, phenol biodegradation using novel bacteria isolated from sewage sludge was investigated. Twenty samples from sewage sludge (SS) were collected, and bacteria in SS contents were cultured in the mineral salt agar (MSA) containing phenol (500 mg/L). Twenty colonies (S1 up to S20) were recovered from all the tested SS samples. The characteristics of three bacterial properties, 16S rDNA sequencing, similarities, GenBank accession number, and phylogenetic analysis showed that strains S3, S10, and S18 were Pseudomonas aeruginosa, Klebsiella pneumoniae, and Klebsiella variicola, respectively. P. aeruginosa, K. pneumoniae, and K. variicola were able to degrade 1000 mg/L phenol in the mineral salt medium. The bacterial st...
2014
A strain of Pseudomonas aeuriginosa MTCC 4997 isolated newly from effluents collected from petrochemical industries. The isolated strain was optimized for various operational and environmental conditions in batch culture. This strain was utilizes phenol as a sole source of carbon and energy, capable of degrading phenol up to 1400 mg l -1 concentration within 144 h. Complete degradation was observed at wide temperature from 15 0 C to 45 0 C with an optimum of 37 0 C and pH 6.0 to 10.5 with an optimum of 7.0 to 7.5. Metals such as Fe, Cd, Cu, Pb, Zn, Mn and Ba at lower concentrations were found stimulated and enhanced the rate of phenol degradation. Therefore, the phenol degradability of the strain can be maintained and used at large scale treatment.
Ecological and physiological analyses of Pseudomonad species within a phenol remediation system
Journal of microbiological …, 2001
A diverse collection of 700 bacteria obtained from an operational phenolic remediating industrial treatment plant was made to select potential strains as microbial biosensors. Pseudomonads were the most abundant group, of which 48 selected from the liquor or suspended solids were assessed for their physiological response to phenolic pollutant loading and niche specialisation. By FAME-MIS identification the Pseudomonads were clustered into six major species groups. Those isolates able to utilise phenol as a sole carbon source predominantly belonged to a non-clonal Pseudomonas pseudoalcaligenes cluster determined by REP-PCR genotyping. Rapid microtitre based respiration assays were developed to contrast activity in response to increasing concentrations of phenol. A considerable range in response for both phenol degrader and non-degrader strains was observed. This natural phenotypic and physiological heterogeneity could facilitate the selection of isolates for the development of a suite of ecologically relevant, custom designed sensors with predictable toxicity susceptibilities to monitor process efficacy. q
Microbial Degradation of Phenol by an Application of Pseudomonas mendocina
Austin Publishing Group
The present investigation was undertaken to isolate and assess the microbial degradation of phenol by bioaugmentation of Pseudomonas mendocina isolated from activated sludge of Common Effluent Treatment Plant (CETP) based at Ankleshwar (Gujarat, India). The strain was isolated and designated as Pseudomonas cepacia ETL 2413 after examined for colony morphology, gram stain characteristics and various biochemical tests. Pseudomonas cepacia ETL 2413 was found to be highly effectual for the removal of phenol which was used as sole carbon and energy source. From an initial concentration of 200mg/L, it degraded to 76.43 ± 1.23 mg/L. In turn the effect of temperature (25 to 500C), pH (5.5 - 10.5) and glucose concentration (0, 0.25 and 0.5%) on the rate of phenol degradation was investigated. Observations revealed that the rate of phenol biodegradation was affected by pH, temperature of incubation and glucose concentration. The optimal conditions for phenol removal were found to be pH of 7.5 (82.63% removal), temperature 300C (78.69% removal) and 0.25% supplemented glucose level (98.28% removal). The significance of the study is the utilization of native bacterial strains isolated from the waste water itself having potential for environmental bioremediation in the activated sludge process of a CETP Plant.
Biodegradation of phenol by free and immobilised cells of locally-isolated bacteria
2017
Phenol is mainly used by the industries to produce a variety of chemical products such as resins, textiles, pesticides, plastics and explosive. Due to the wider use of phenol and other phenolic compounds by industries, this has resulted in an increased presence of these toxic compounds in the environment. In Malaysia, phenol and phenolic compounds rank among the top three scheduled wastes with thousands of tonnes being produced yearly for disposal. In Malaysia about 37.7 metric tonnes of phenol and phenol-containing wastes are produced in 2014, there is also an incident of tanker accidents Straits of Malacca in 2003 where tonnes of phenol spilt into the river and the Kapar power plant in Klang, Selangor uses coal thereby producing phenol as a by-product.These make phenol one of the environmental problem in Malaysia. Bioremoval of phenol by microorganisms especially bacteria has been demonstrated to be the most effective and economical approach compared to physio-chemical methods. The search for efficient phenol-degraders especially local sources to remediate local phenol pollution is important as indigenous bacteria usually have better survival and resilient to local geographical conditions. In this study, phenol-degrading microorganisms were isolated from local soils and water bodies. Identification was carried out using 16s rRNA gene sequencing and molecular phylogeny analysis using the Phylip software. The isolates were inoculated in mineral salt media with 0.5 g/L phenol as the sole source of carbon. Phenol degradation was determined using 4-amino antipyrine method. Physical and cultural conditions influencing phenol degradation such as pH, temperature, nature of bacteria, salinity, and nitrogen source were optimised via one-factor-at-a-time and response surface methodology (RSM). The robust and hardy Gellan gum was used for the immobilisation of bacterial cells and also the ortho and meta-pathways for phenol degradation were elucidated. The highest degradation was achieved at pH 7.5 (phosphate buffer) for all of the three isolates, with an optimum temperature of 30°C for Pseudomonas sp. AQ5-04 and Alcaligenes sp. AQ5-02 and 32.5°C for Serratia sp. AQ5-03. Ammonium sulphate was established 04 showing the best reusability of 22 cycles compared to 16 and 14 cycles for Alcaligenes sp. AQ5-02 and Serratia sp. AQ5-03, respectively. The immobilised cell of Alcaligenes sp. AQ5-02, Serratia sp. AQ5-03 and Pseudomonas sp. AQ5-04 can degrade up to 1900 mg/L. All the three isolates have the ability to degrade phenol both in free and immobilised cells. Immobilisation has significantly enhanced their biodegradation ability. Pseudomonas sp. AQ5-04 has the highest reusability as well as tolerating slightly high salinity. The meta pathway for phenol degradation was detected for Alcaligenes sp. AQ5-02 Pseudomonas sp. AQ5-04 while the ortho pathway was detected for, Serratia sp. AQ5-03. The accuracy and statistical analysis of the kinetic models used show that the best model was Luong for all bacterial growth curves with the lowest values for root mean square error or RMSE and adjusted Akaike Information criteria AICc, highest adjusted R 2 values, and with Bias Factor and Accuracy Factor nearest to unity (1.0) for Pseudomonas sp. AQ05-04 and Serratia sp. AQ05-03, with the exception of Alcaligenes sp. AQ05-02 where the AICc value was not the lowest but the rest of the statistical analysis values still overwhelmingly pinpointing the Luong model as the best model for Alcaligenes sp. AQ05-02. The calculated value for the Luong's constants maximal growth rate, half saturation constant for maximal growth, maximal concentration of substrate tolerated and curve parameter that defines the steepness of the growth rate decline from the maximum rate, symbolized by umax, Ks, Sm, and n were 0.10±0.02 hr-1 , 0.02±0.01 g/L, 2.05±0.06 g/L and 0.80±0.20 (± 95% confidence interval) for Pseudomonas sp. AQ05-04, 0.07±0.02 hr-1 , 0.02±0.01 g/L, 1.18±0.03 g/L and 1.16±0.23 for Serratia sp.AQ05-03, and 0.07±0.01 hr-1 , 0.18±0.03 g/L, 1.27±0.24 g/L and 6.60±0.94 for Alcaligenes sp. AQ05-02, respectively. It appears that the highest maximum growth rate on phenol was exhibited by Pseudomonas sp. AQ05-04, while both Serratia sp.AQ05-03 and Alcaligenes sp. AQ05-02 had similar lower growth rates indicating that Pseudomonas sp. AQ05-04 showed a higher efficient growth rate on phenol.
2021
This study examines the isolation and characterization of phenol-degrading-bacteria, Five isolates named C2, P4, M4, O3, and S3, from the effluent of ceramic factories (C2), petrochemical (P4), the pulp paper mill (M4), crude oil (O3) and sewage water (S3) have been investigated. These isolates were characterized depending on their morphological, and biochemical characteristics, only one isolate was characterized as a Gram-positive, strictly aerobic, nonmotile, and cocci-shaped bacterium might be Micrococcus sp. (M4), two isolates (Pseudomonas sp.: (P4&S3) were recogniced as a Gram-negative, strictly aerobic, motile and short rod-shaped bacterium and two isolates(Bacillus sp.: (C2& O3) were idenihied as a Gram-positive, strictly aerobic, motile and long rod- shape bacterium. The five bacterial isolates were able to utilize phenol as a sole carbon source. These isolates were checked for growth on a minimal salt medium amended with different concentrations of phenol. The five tolerant...
Characterization and identification of phenol degrading bacteria isolated from industrial waste
Pakistan Journal of Agricultural Sciences
Phenol is a toxic organic pollutant to living cells and its biodegradation is considered the best method due to its environment friendly nature and cost effectiveness. In this study, eight bacterial strains were isolated through enrichment on mineral salt media supplemented with 300 mg L-1 phenol. The isolated strains were identified by 16S rRNA gene sequence analysis and belonged to genera: Rhodococcus, Stenotrophomonas, Lysinibacillus, Comamonas, Microbacterium, Pseudomonas and Halomonas. The results of phenol biodegradation experiments (conducted at pH 7 and 30°C temperature) showed that the strains could degrade 750 mg L-1 phenol within 40 to 96 hours. The average phenol degradation rate by the strains was 12.5 to 34.8 mgL-1h-1. The most rapid phenol degradation was observed for Rhodococcus sp. NCCP-309 and Rhodococcus sp. NCCP-312, whereas, Stenotrophomonas sp. NCCP-311, Lysinibacillus sp. NCCP-313, Comamonas sp. NCCP-314 and Microbacterium sp. NCCP-351 took longer time in phen...