Abstract
Pseudomonas aeruginosa PP4, Pseudomonas sp. PPD and Acinetobacter lwoffii ISP4 capable of utilizing phthalate isomers were isolated from the soil using enrichment culture technique. The strain ISP4 metabolizes isophthalate, while PPD and PP4 utilizes all three phthalate isomers (ortho-, iso- and tere-) as the sole carbon source. ISP4 utilizes isophthalate (0.1%) more rapidly (doubling time, 0.9 h) compared to PPD (4.64 h), PP4 (7.91 h) and other reported strains so far. The metabolic pathways in these isolates were initiated by dihydroxylation of phthalate isomers. Phthalate is hydroxylated to 3,4-dihydro-3,4-dihydroxyphthalate and 4,5-dihydro-4,5-dihydroxyphthalate in strains PP4 and PPD, respectively; while terephthalate is hydroxylated to 2-hydro-1,2-dihydroxyterephthalate. All three strains hydroxylate isophthalate to 4-hydro-3,4-dihydroxyisophthalate. The generated dihydroxyphthalates were subsequently metabolized to 3,4-dihydroxybenzoate (3,4-DHB) which was further metabolized by ortho ring-cleavage pathway. PP4 and PPD cells grown on phthalate, isophthalate or terephthalate showed respiration on respective phthalate isomer and the activity of corresponding ring-hydroxylating dioxygenase, suggesting the carbon source specific induction of three different ring-hydroxylating dioxygenases. We report, for the first time, the activity of isophthalate dioxygenase and its reductase component in the cell-free extracts. The enzyme showed maximum activity with reduced nicotinamide adenine dinucleotide (NADH) in the pH range 8–8.5. Cells grown on glucose failed to respire on phthalate isomers and 3,4-DHB and showed significantly low activities of the enzymes suggesting that the enzymes are inducible.
References
Arciero DM, Orville AM, Lipscomb JD (1990) Protocatechuate 4,5-dioxygenase from Pseudomonas testosteroni. Methods Enzymol 188:89–95
Basu A, Dixit SS, Phale PS (2003) Metabolism of benzyl alcohol via catechol ortho-pathway in methylnaphthalene-degrading Pseudomonas putida CSV86. Appl Microbiol Biotechnol 62:579–585
Batie CJ, LaHaie E, Ballou DP (1987) Purification and characterization of phthalate oxygenase and phthalate oxygenase reductase from Pseudomonas cepacia. J Biol Chem 262:1510–1518
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem 72:248–254
Choi KY, Kim D, Sul WJ, Chae JC, Zylstra GJ, Kim YM, Kim E (2005) Molecular and biochemical analysis of phthalate and terephthalate degradation by Rhodococcus sp strain DK17. FEMS Microbiol Lett 252:207–213
Correll CC, Batie CJ, Ballou DP, Ludwig ML (1992) Phthalate dioxygenase reductase: a modular structure for electron transfer from pyridine nucleotides to [2Fe–2S]. Science 258:1604–1610
Dillingham EO, Autian J (1973) Teratogenicity, mutagenicity and cellular toxicity of phthalate esters. Environ Health Perspect 3:81–89
Eaton RW (2001) Plasmid-encoded phthalate catabolic pathway in Arthrobacter keyseri 12B. J Bacteriol 183:3689–3703
Fujisawa H (1970) Protocatechuate 3,4-dioxygenase (Pseudomonas). Methods Enzymol 17A:526–529
Gassner GT, Ballou DP (1995) Preparation and characterization of a truncated form of phthalate dioxygenase reductase that lacks an iron–sulfur domain. Biochemistry 34:13460–13471
Gesler RM (1973) Toxicology of di-2-ethylhexyl phthalate and other phthalic acid ester plasticizers. Environ Health Perspect 3:73–79
Gibson DT, Subramanian V (1984) Microbial degradation of phthalates. In: Gibson DT (ed) Microbial degradation of organic compounds. Dekker, New York, pp 371–394
Harris CA, Henttu P, Parker MG, Sumpter JP (1997) The estrogenic activity of phthalate esters in vitro. Environ Health Perspect 105:802–811
Holt JG, Krieg NR, Sneath PHA, Staley JT, Williams ST (2000) Bergey’s manual of determinative bacteriology, 9th edn. Lippincott Williams and Wilkins, Philadelphia, PA
Jaeger RJ, Rubin RJ (1973) Extraction, localization, and metabolism of di-2-ethylhexyl phthalate from PVC plastic medical devices. Environ Health Perspect 3:95–102
Kaun-Yu L, Fu-Wei T, Chia-Jung W, Pei-Shan L (2004) Suppression by phthalates of the calcium signaling of human nicotinic acetylcholine receptors in human neuroblastoma SH-SY5Y cells. Toxicology 200:113–121
Keyser P, Pujar BG, Eaton RW, Ribbons DW (1976) Biodegradation of the phthalates and their esters by bacteria. Environ Health Perspect 18:159–166
Koch HM, Drexler H, Angerer J (2003) An estimation of the daily intake of di(2-ethylhexyl)phthalate (DEHP) and other phthalates in the general population. Int J Hyg Environ Health 206:77–83
Krauskopf LG (1973) Studies on the toxicity of phthalates via ingestion. Environ Health Perspect 3:61–72
Maruyama K, Akita K, Naitou C, Yoshida M, Kitamura T (2005) Purification and characterization of an esterase hydrolyzing monoalkyl phthalates from Micrococcus sp. YGJ1. J Biochem 137:27–32
Niazi JH, Prasad DT, Karegoudar TB (2001) Initial degradation of dimethylphthalate by esterases from Bacillus species. FEMS Microbiol Lett 196:201–205
Nomura Y, Harashima S, Oshima Y (1989) A simple method for detection of enzyme activities involved in the initial step of phthalate degradation in microorganisms. J Ferment Bioeng 67:291–296
Phale PS, Mahajan MC, Vaidyanathan CS (1995) A pathway for biodegradation of 1-naphthoic acid by Pseudomonas maltophilia CSV89. Arch Microbiol 163:42–47
Pujar BG, Ribbons DW (1985) Phthalate-metabolism in Pseudomonas fluorescens PHK: Purification and properties of 4,5-dihydroxyphthalate decarboxylase. Appl Environ Microbiol 49:374–376
Rani M, Prakash D, Sobti RC, Jain RK (1996) Plasmid mediated degradation of o-phthalate and salicylate by a Moraxella sp. Biochem Biophys Res Commun 220:377–381
Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbour Laboratory, NY
Schlafli HR, Weiss MA, Leisinger T, Cook AM (1994) Terephthalate 1,2-dioxygenase system from Comamonas testosteroni T-2: purification and some properties of the oxygenase component. J Bacteriol 176:6644–6652
Shigematsu T, Yumihara K, Ueda Y, Morimura S, Kida K (2003) Purification and gene cloning of the oxygenase component of the terephthalate 1,2-dioxygenase system from Delftia tsuruhatensis strain T7. FEMS Microbiol Lett 220:255–260
Swetha VP, Phale PS (2005) Metabolism of carbaryl via 1,2-dihydroxy naphthalene by soil isolates Pseudomonas sp. strains C4, C5 and C6. Appl Environ Microbiol 71:5951–5956
Tarasev M, Ballou DP (2005) Chemistry of the catalytic conversion of phthalate into its cis-dihydrodiol during the reaction of oxygen with the reduced form of phthalate dioxygenase. Biochemistry 44:6197–6207
Vega D, Bastide J (2003) Dimethylphthalate hydrolysis by specific microbial esterase. Chemosphere 51:663–668
Wang YZ, Zhou Y, Zylstra GJ (1995) Molecular analysis of isophthalate and terephthalate degradation by Comamonas testosteroni YZW-D. Environ Health Perspect 103:9–12
Acknowledgements
Research grant from the Department of Biotechnology, Govt. of India to PP and CSIR-Junior research fellowship to CVK is gratefully acknowledged. Thanks to Dr. Shouche Y, NCCS, India for the 16S rRNA sequencing and Mr. Prabin K Majhi for constructive discussion.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Vamsee-Krishna, C., Mohan, Y. & Phale, P.S. Biodegradation of Phthalate Isomers by Pseudomonas aeruginosa PP4, Pseudomonas sp. PPD and Acinetobacter lwoffii ISP4. Appl Microbiol Biotechnol 72, 1263–1269 (2006). https://doi.org/10.1007/s00253-006-0413-7
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00253-006-0413-7