Characterization of the Phenol-degrading Pseudomonas sp. strain Neni-4
DOI:
https://doi.org/10.54987/jebat.v7i1.1044Keywords:
Pseudomonas, Phenol biodegradation, Meta pathway, Heavy metals, BioremediationAbstract
This research presents the characterization of Pseudomonas sp. strain Neni-4 , which was selected because it can metabolize phenol as its primary carbon source. The strain breaks down phenol through the meta-cleavage pathway , which is typical for many efficient phenol-degrading bacteria. The strain achieved its highest growth rate at temperatures ranging from 25 to 30°C and pH values between 7.0 and 7.5 without significant differences (p > 0.05) in these ranges. The bacterial growth reached its peak when phenol concentrations ranged from 500 to 700 mg/L. The best growth results occurred when using 1% (w/v) ammonium sulfate as the nitrogen source. The research investigated the impact of heavy metals on the phenol breakdown process. Cd(II) showed no inhibitory effect on the process , and similarly, Pb(II) and Ni(II) showed more than 90% activity was observed. The growth rate decreased by 27% when exposed to Cu(II), but Ag(I) and Hg(II) caused extreme inhibition, which reduced activity to 12% and 5% respectively. The degradation profile indicated a rapid decrease of phenol concentration from 500 mg/L to near-zero levels during the first five days, while the cell density increased, indicating active biodegradation. Pseudomonas sp. strain Neni-4 demonstrates potential for bioremediation of phenol-contaminated sites when operated under neutral pH and mesophilic temperature conditions.
References
Acosta CA, Pasquali CEL, Paniagua G, Garcinuño RM, Hernando PF. Evaluation of total phenol pollution in water of San Martin Canal from Santiago del Estero, Argentina. Environ Pollut. 2018;236:265-72.
Beelen P van, Fleuren-Kemilä AK, Huys MPA, Montforta ACP van, Vlaardingen PLA van. The toxic effects of pollutants on the mineralization of acetate in subsoil microcosms. Environ Toxicol Chem. 1991;10(6):775-89.
Chun UH, Simonov N, Chen Y, Britz ML. Continuous pollution monitoring using Photobacterium phosphoreum. Resour Conserv Recycl. 1996;18(1-4):25-40.
Bosco ML, Varrica D, Dongarrà G. Case study: Inorganic pollutants associated with particulate matter from an area near a petrochemical plant. Environ Res. 2005;99(1):18-30.
Agarry S e., Durojaiye A o., Yusuf R o., Aremu M o., Solomon B o., Mojeed O. Biodegradation of phenol in refinery wastewater by pure cultures of Pseudomonas aeruginosa NCIB 950 and Pseudomonas fluorescence NCIB 3756. Int J Environ Pollut. 2008 Jan;32(1):3-11.
Abd El-Zaher EHF, Mahmoud YAG, Aly MM. Effect of different concentrations of phenol on growth of some fungi isolated from contaminated soil. Vol. 10, African Journal of Biotechnology. 2011. p. 1384-92.
Gami AA, Shukor MY, Khalil KA, Dahalan FA, Khalid A, Ahmad SA. Phenol and its toxicity. J Environ Microbiol Toxicol. 2014;2(1):11-23.
Mohanta V, Mishra B. Monitoring of phenol in river and groundwater of adjoining area of steel city: A case study of Burnpur, West Bengal. 2018.
Sukumaran D. A Preliminary Study on the Estimation of Phenol content and Extent of Pollution in water bodies near the Plywood Industrial Area, Perumbavoor, Ernakulum (Dist), Kerala, India. Asian J Res Chem. 2022 Oct 3;15(5): September-October 2022:311-3.
Mohanta VL, Mishra BK. Occurrence and fate of phenolic compounds in groundwater and their associated risks. In: Legacy, Pathogenic and Emerging Contaminants in the Environment. CRC Press; 2021.
Basak B, Jeon BH, Kurade MB, Saratale GD, Bhunia B, Chatterjee PK, et al. Biodegradation of high concentration phenol using sugarcane bagasse immobilized Candida tropicalis PHB5 in a packed-bed column reactor. Ecotoxicol Environ Saf. 2019 Sept 30;180:317-25.
Zhao T, Gao Y, Yu T, Zhang Y, Zhang Z, Zhang L, et al. Biodegradation of phenol by a highly tolerant strain Rhodococcus ruber C1: Biochemical characterization and comparative genome analysis. Ecotoxicol Environ Saf. 2021 Jan 15;208:111709.
Zhang S, An Z, Su X, Hou A, Liu L, Zhang L, et al. Phenol degradation at high salinity by a resuscitated strain Pseudomonas sp. SAS26: kinetics and pathway. J Environ Chem Eng. 2023 Aug 1;11(4):110182.
Huang L, Liang J, Zhang J. Characteristics of a novel acid-resistant phenol-degrading bacterium Acinetobacter pittii Hly3: Adaptability, kinetics, degradation pathway and long-term performance. Int Biodeterior Biodegrad. 2024 July 1;192:105825.
González G, Herrera G, García MaT, Peña M. Biodegradation of phenolic industrial wastewater in a fluidized bed bioreactor with immobilized cells of Pseudomonas putida. Vol. 80, Bioresource Technology. 2001. p. 137-42.
Kumar A, Kumar S, Kumar S. Biodegradation kinetics of phenol and catechol using Pseudomonas putida MTCC 1194. Vol. 22, Biochemical Engineering Journal. 2005. p. 151-9.
Afzal M, Iqbal S, Rauf S, Khalid ZM. Characteristics of phenol biodegradation in saline solutions by monocultures of Pseudomonas aeruginosa and Pseudomonas pseudomallei. Vol. 149, Journal of Hazardous Materials. 2007. p. 60-6.
Kotresha D, Vidyasagar GM. Isolation and characterisation of phenol-degrading Pseudomonas aeruginosa MTCC 4996. Vol. 24, World Journal of Microbiology and Biotechnology. 2008. p. 541-7.
El Deeb B, Altalhi AD. Degradative plasmid and heavy metal resistance plasmid naturally coexist in phenol and cyanide assimilating bacteria. Vol. 5, American Journal of Biochemistry and Biotechnology. 2009. p. 84-93.
Singh U, Arora NK, Sachan P. Simultaneous biodegradation of phenol and cyanide present in coke-oven effluent using immobilized Pseudomonas putida and Pseudomonas stutzeri. Braz J Microbiol. 2018 Jan 1;49(1):38-44.
Uba G, Yakasai HM, Abubakar A. Mathematical modeling of the biodegradation of phenol from industrial effluents using immobilized Pseudomonas putida. J Biochem Microbiol Biotechnol. 2020 July 31;8(1):15-8.
Abubakar A, Yakasai HM, Uba G, Sabo I. Substrate Inhibition Modelling of Pseudomonas nitroreducens Growth on Octylphenol Polyethoxylates. J Environ Microbiol Toxicol. 2023 June 30;11(1):25-31.
Kennes C, Lema JM. Degradation of major compounds of creosotes (PAH and phenols) by Phanerochaete chrysosporium. Vol. 16, Biotechnology Letters. 1994. p. 759-64.
Leitão AL, Duarte MP, Oliveira JS. Degradation of phenol by a halotolerant strain of Penicillium chrysogenum. Vol. 59, International Biodeterioration and Biodegradation. 2007. p. 220-5.
Yan J, Jianping W, Jing B, Daoquan W, Zongding H. Phenol biodegradation by the yeast Candida tropicalis in the presence of m-cresol. Vol. 29, Biochemical Engineering Journal. 2006. p. 227-34.
dos Santos VL, Monteiro A de S, Braga DT, Santoro MM. Phenol degradation by Aureobasidium pullulans FE13 isolated from industrial effluents. Vol. 161, Journal of Hazardous Materials. 2009. p. 1413-20.
Arutchelvan V, Kanakasabai V, Elangovan R, Nagarajan S, Muralikrishnan V. Kinetics of high strength phenol degradation using Bacillus brevis. Vol. 129, Journal of Hazardous Materials. 2006. p. 216-22.
Nair IC, Jayachandran K, Shashidhar S. Treatment of paper factory effluent using a phenol degrading Alcaligenes sp. under free and immobilized conditions. Vol. 98, Bioresource Technology. 2007. p. 714-6.
Kiliç NK. Enhancement of phenol biodegradation by Ochrobactrum sp. isolated from industrial wastewaters. Vol. 63, International Biodeterioration and Biodegradation. 2009. p. 778-81.
Jones AL, Brown JM, Mishra V, Perry JD, Steigerwalt AG, Goodfellow M. Rhodococcus gordoniae sp. nov., an actinomycete isolated from clinical material and phenol-contaminated soil. Vol. 54, International Journal of Systematic and Evolutionary Microbiology. 2004. p. 407-11.
Rehfuss M, Urban J. Rhodococcus phenolicus sp. nov., a novel bioprocessor isolated actinomycete with the ability to degrade chlorobenzene, dichlorobenzene and phenol as sole carbon sources. Vol. 28, Systematic and Applied Microbiology. 2005. p. 695-701.
Nagamani A, Lowry M. Phenol biodegradation by Rhodococcus coprophilus isolated from semi arid soil samples of Pali, Rajasthan. Vol. 4, International Journal of Applied Environmental Sciences. 2009. p. 295-302.
Shumkova ES, Solyanikova IP, Plotnikova EG, Golovleva LA. Phenol degradation by Rhodococcus opacus strain 1G. Vol. 45, Applied Biochemistry and Microbiology. 2009. p. 43-9.
Arif NM, Ahmad SA, Syed MA, Shukor MY. Isolation and characterization of a phenol-degrading Rhodococcus sp. strain AQ5NOL 2 KCTC 11961BP. J Basic Microbiol. 2013;53(1):9-19.
Pinto G, Pollio A, Previtera L, Temussi F. Biodegradation of phenols by microalgae. Vol. 24, Biotechnology Letters. 2002. p. 2047-51.
Souvannachith T, Putra D, Hendrayana H. Assessment of Groundwater Contamination Hazard by Nitrate in Samas Area, Bantul District, Yogyakarta, Indonesia. J Appl Geol. 2017 Nov 13;2:36.
Rusnam, Gusmanizar N, Rahman MF, Yasid NA. Characterization of a Molybdenum-reducing and Phenol-degrading Pseudomonas sp. strain Neni-4 from soils in West Sumatera, Indonesia. Bull Environ Sci Sustain Manag. 2022 July 31;6(1):1-8.
Hughes EJL, Bayly RC, Skurray RA. Evidence of isofunctional enzymes in the degeneration of phenol, m- and p- toluate, and p-cresol via catechol meta-cleavage pathways in Alcaligenes eutrophus. Vol. 158, Journal of Bacteriology. 1984. p. 79-83.
Powlowski J, Shingler V. Genetics and biochemistry of phenol degradation by Pseudomonas sp. CF600. Vol. 5, Biodegradation. 1994. p. 219-36.
El-Sayed WS a, Ibrahim MK a, Abu-Shady M a, El-Beih F a, Ohmura N b, Saiki H b, et al. Isolation and characterization of phenol-catabolizing bacteria from a coking plant. Biosci Biotechnol Biochem. 2003;67(9):2026-9.
Yang CF, Lee CM. Enrichment, isolation, and characterization of phenol-degrading Pseudomonas resinovorans strain P-1 and Brevibacillus sp. strain P-6. Vol. 59, International Biodeterioration and Biodegradation. 2007. p. 206-10.
Basak SP, Sarkar P, Pal P. Isolation and characterization of phenol utilizing bacteria from industrial effluent-contaminated soil and kinetic evaluation of their biodegradation potential. J Environ Sci Health - Part ToxicHazardous Subst Environ Eng. 2014;49(1):67-77.
Kujur RRA, Das SK. Pseudomonas phenolilytica sp. nov., a novel phenol-degrading bacterium. Arch Microbiol. 2022 May 14;204(6):320.
Oh JS a, Han YH b. Isolation and Characterization of Phenol-degrading Rhodococcus sp. DGUM 2011. Korean J Appl Microbiol Biotechnol. 1997;25(5):459-63.
Kim D, Kim YS, Kim SK, Kim SW, Zylstra GJ, Kim YM, et al. Monocyclic aromatic hydrocarbon degradation by Rhodococcus sp. strain DK17. Appl Environ Microbiol. 2002;68(7):3270-8.
Zaki S. Detection of meta - and ortho -cleavage dioxygenases in bacterial phenol-degraders. J Appl Sci Environ Manag. 2006 Oct 31;10(3):75-81.
Alkafaween MA, Magharbeh MK, Khleifat KM, Saraireh R, Qaralleh H, El-Hasan T, et al. Biodegradation of Phenol by Bacillus simplex: Characterization and Kinetics Study. Appl Environ Biotechnol. 2021 Nov 29;6(2):1-12.
Benkhennouche-Bouchene H, Mahy JG, Lambert SD, Hayoun B, Deflaoui O, Bourouina M, et al. Statistical modeling and optimization of Escherichia coli growth parameters for the biological treatment of phenol. Biocatal Agric Biotechnol. 2021 July 1;34:102016.
Magharbeh MK, Khleifat KM, Al-Kafaween MA, Saraireh R, Alqaraleh M, Qaralleh H, et al. Biodegradation of phenol by Bacillus simplex: Characterization and kinetics study. Appl Environ Biotechnol. 2021;6(2):1-12.
Allamin IA, Ya'u M, Muhammad UA, Abubakar A. Substrate Inhibition Kinetics Models for Fitting the Growth Rate of Phenol by an Acclimatized Mixed Bacterial Consortia from an Anaerobic Batch Reactor. Bull Environ Sci Sustain Manag. 2023 July 31;7(1):44-51.
Mortazavi A, Hassanshahian M, Ali E, Fenjan MN, Alawadi A, Alsalamy A. Isolation and Identification of Phenol-Degrading Bacteria from Iranian Soil and Leaf Samples. Front Biosci-Elite. 2023 Dec 12;15(4):29.
Rusnam, Rahim MBHA, Rahman MF, Khayat ME, Nasution FI, Yakasai HM. Primary Mathematical Modeling of Growth on Phenol by Bacillus sp. Strain Neni-10. J Environ Bioremediation Toxicol. 2023 Dec 31;6(2):29-36.
Szilveszter S, Fikó DR, Máthé I, Felföldi T, Ráduly B. Kinetic characterization of a new phenol degrading Acinetobacter towneri strain isolated from landfill leachate treating bioreactor. World J Microbiol Biotechnol. 2023 Jan 17;39(3):79.
Szilveszter S, Fikó DR, Máthé I, Felföldi T, Ráduly B. Kinetic characterization of a new phenol degrading Acinetobacter towneri strain isolated from landfill leachate treating bioreactor. World J Microbiol Biotechnol. 2023 Jan 17;39(3):79.
Husein N, Qaralleh H, Al-Tarawneh A, AlSarayreh A, Qaisi YA, Al-limoun M, et al. Modeling phenol biodegradation with Pantoea agglomerans as plant-growth-promoting bacteria J Adv Pharm Educ Res. 2024;14(2-2024):63-71.
Shebl S, Ghareeb DA, Ali SM, Ghanem NBED, Olama ZA. Aerobic phenol degradation using native bacterial consortium via ortho-and meta-cleavage pathways. Front Microbiol [Internet]. 2024 Aug 5 [cited 2025 Mar 10];15. Available from: https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2024.1400033/full
Buswell JA. Metabolism of phenol and cresols by Bacillus stearothermophilus. J Bacteriol. 1975 Dec;124(3):1077-83.
Fernández PM, Martorell MM, Blaser MG, Ruberto LAM, de Figueroa LIC, Mac Cormack WP. Phenol degradation and heavy metal tolerance of Antarctic yeasts. Extrem Life Extreme Cond. 2017 May;21(3):445-57.
Lee GLY, Yasid NA, W-Johari WL, Zulkharnain A, Alias SA, Shukor MY, et al. Screening and isolation of Antarctic phenol-degrading bacteria. In 10-3 Midori-cho, Tachikawa, Tokyo, Japan: National Institute of Polar Research (NIPR); 2018. Available from: http://id.nii.ac.jp/1291/00013651/
Al-Khalid T, El-Naas MH. Aerobic biodegradation of phenols: A comprehensive review. Crit Rev Environ Sci Technol. 2012;42(16):1631-90.
Harianja S, Wirawan IG, Sritamin M. Isolation and ability test of phenol-degrading bacteria from the liquid waste in bhayangkara hospital, denpasar city, bali, indonesia. Int J Biosci Biotechnol. 2021 Sept 30;9:49.
Hanafee N, Salleh NAM, Ahmad SA, Saada WZ, Yusof MT. Characterization of phenol-degrading fungi isolated from industrial waste water in Malaysia. Asia-Pac J Mol Biol Biotechnol. 2019;27(2):35-43.
Wang L, Li Y, Niu L, Dai Y, Wu Y, Wang Q. Isolation and growth kinetics of a novel phenol-degrading bacterium Microbacterium oxydans from the sediment of Taihu Lake (China). Water Sci Technol. 2016;73(8):1882-90.
Heilbuth NM, Linardi VR, Monteiro AS, da RRA, Mimim LA, Santos VL. Estimation of kinetic parameters of phenol degradation by bacteria isolated from activated sludge using a genetic algorithm. J Chem Technol Biotechnol. 2015;90(11):2066-75.
Bai J, Wen JP, Li HM, Jiang Y. Kinetic modeling of growth and biodegradation of phenol and m-cresol using Alcaligenes faecalis. Vol. 42, Process Biochemistry. 2007. p. 510-7.
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