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Purification and characterization of a lyase from the EDTA-degrading bacterial strain DSM 9103 that catalyzes the splitting of [S,S]-ethylenediaminedisuccinate, a structural isomer of EDTA

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Abstract

The bacterial strain DSM 9103, able to utilize EDTA as a sole source of carbon, nitrogen, and energy, is also capable to grow with [S,S]-ethylenediaminedisuccinate ([S,S]-EDDS), a structural isomer of EDTA. In cell-free extracts of[S,S]-EDDS-grown bacteria, [S,S]-EDDS degradation was observed in the absence of any cofactors. An enzyme was purified41-fold that catalyzed the non-hydrolytic splitting of[S,S]-EDDS leading to the formation of fumarate and N-(2-aminoethyl) aspartic acid. These data strongly suggest that the enzyme belongs to the group of carbon-nitrogen lyases. The splitting reaction was reversible, and an equilibrium constant of approximately 43.0 10-1 M was determined. Out of the three stereo-isomers of EDDS, [S,S]-and [R,S]-EDDS were accepted as substrates by the lyase,whereas [R,R]-EDDS remained unchanged in assays with both cell-free extracts and pure enzyme. The enzyme catalyzed the transformation of free [S,S]-EDDS and of [S,S]-EDDS-metal complexes with stability constant lower than 10, namely of MgEDDS, CaEDDS, BaEDDS and to a small extent also of MnEDDS;FeIIIEDDS, NiEDDS, CuEDDS, CoEDDS and ZnEDDS were not transformed.

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References

  • Alder AC, Siegrist H, Gujer W & Giger W (1990) Behaviour of NTA and EDTA in biological wastewater treatment. Wat. Res. 24: 733–742

    Article  CAS  Google Scholar 

  • Auling G, Busse H-J, Egli T, El-Banna T & Stackebrandt E (1993) Description of the Gram-negative, obligately aerobic, nitrilotriacetate (NTA)-utilizing bacteria as Chelatobacter heintzii, gen. nov., sp. nov., and Chelatococcus asaccharovorans, gen. nov., sp. nov. Syst. Appl. Microbiol. 16: 104–112

    Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantifi-cation of microgram quantities of proteins utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248–254

    Article  CAS  Google Scholar 

  • Cebulla I (1995) Gewinnung komplexbildender Substanzen mittels Amycolatopsis orientalis. PhD thesis, Eberhard-Karls-Universität Tübingen, Germany

    Google Scholar 

  • Dietz F (1987) Neue Meßergebnisse über die Belastung von Trinkwasser mit EDTA. gwf Wasser Abwasser 128: 286–288

    CAS  Google Scholar 

  • Egli T (1988) (An)aerobic breakdown of chelating agents used in household detergents. Microbiol. Sci. 5: 36–41

    CAS  Google Scholar 

  • Egli T, Weilenmann HU, El-Banna T & Auling G (1988) Gram-negative, aerobic, nitrilotriacetate-utilizing bacteria from wastewater and soil. Syst. Appl. Microbiol. 10: 297–305

    Google Scholar 

  • Frimmel FH, Grenz R, Kordik E & Dietz F (1989) Nitrilotriacetat (NTA) und Ethylendinitrilotetraacetat (EDTA) in Fließgewässern der Bundesrepublik Deutschland. Vom Wasser 72: 175–184

    CAS  Google Scholar 

  • Gorelov IP, Samsonov AP, Nikol'skii VM, Babich VA, Svetogorov YE, Smirnova TI, Malakhaev ED, Kozlov YM & Kapustnikov AI (1978) Synthesis and complex-forming properties of complexons derived from dicarboxylic acids. V. Synthesis of complexons derived from succinic acid. Russ. J. Gen. Chem. 49: 659–663

    Google Scholar 

  • Hartman FA & Perkins CM (1987) Detergent compositions containing ethylenediamine-N,N'-disuccinic acid. U.S. patent 4,704,233

  • Houriet J-P (1996) NTA dans les eaux. Cahier de l'environnement 264. Office fédéral de l'environnement, des forêts et du paysage (OFEFP), Berne, Switzerland

    Google Scholar 

  • Kari FG (1994) Umweltverhalten von Ethylendiamintetraacetat (EDTA) unter spezieller Berücksichtigung des photochemischen Abbaus. PhD thesis No. 10698, Swiss Federal Institute of Technology, Zürich, Switzerland

    Google Scholar 

  • Kovaleva IB, Mitrofanova ND & Martynenko LI (1992) The structure of transition metal complexonates derived from ethylenediamine-(N,N')-disuccinic acid. Russ. J. Inorg. Chem. 37: 41–45

    Google Scholar 

  • Lämmli UK (1970) Cleavage of structural proteins during the assambly of the head of bacteriophage T4. Nature 227: 680–685

    Article  Google Scholar 

  • Ludwig P, Gunkel W & Hühnerfuss H (1992) Chromatographic separation of the enantiomers of marine pollutants. Part 5: Enantioselective degradation of phenoxycarboxylic acid herbicides by marine microorganisms. Chemosphere 24: 1423–1429

    Article  CAS  Google Scholar 

  • Martell AE & Smith RM (1974) Critical stability constants, Vol 1: Amino acids. Plenum Press, New York

    Google Scholar 

  • Neal JA & Rose NJ (1968) Stereospecific ligands and their complexes. I. A cobalt(III)complex of ethylenediaminedisuccinic acid. Inorg. Chem. 7: 2405–2412

    Article  CAS  Google Scholar 

  • Nickel K, Suter MJ-F & Kohler H-P (1997) Involvement of two α-ketoglutarate-dependent dioxygenases in the enantioselective degradation of (R)-and (S)-mecoprop by Sphinogomonas herbicidovorans MH. J. Bacteriol. 179: 6674–6679

    CAS  Google Scholar 

  • Nishikiori T, Okuyama A, Naganawa T, Takita T, Hamada M, Takeuchi T, Aoyagi T & Umezawa H (1984) Production by actinomycetes of (S,S)-N,N'-ethylenediaminedisuccinic acid, an inhibitor of phospholipase C. J. Antibiot. 37: 426–427

    CAS  Google Scholar 

  • Nowack B (1996) Behaviour of EDTA in groundwater-a study of the surface reactions of metal-EDTA complexes. PhD thesis No. 11392, Swiss Federal Institute of Technology, Zürich, Switzerland

    Google Scholar 

  • Pavelcik F, Garaj J & Majer J (1980) Lithium [(R,S)-N,N'-ethylenediaminedisuccinato]cobaltate(III) trihydrate. Acta Chryst. B36: 2152–2154

    Article  CAS  Google Scholar 

  • Schneider RP, Zürcher F, Egli T & Hamer G (1988) Determination of nitrilotriacetate in biological matrices using ion exclusion chromatography. Anal. Biochem. 173: 278–284

    Article  CAS  Google Scholar 

  • Schowanek D, Feijtel TCJ, Perkins CM, Hartman FA, Federle TW & Larson RJ (1997) Biodegradation of [S,S], [R,R] and mixed stereoisomers of ethylene diamine disuccinic acid (EDDS), a transition metal chelator. Chemosphere 34: 2375–2391

    Article  CAS  Google Scholar 

  • Simoni S, Klinke S, Zipper C, Angst W & Kohler H-P (1996) Enantioselective metabolism of chiral 3-phenylbutyric acid, an intermediate of linear alkylbenzene degradation, by Rhodococcus rhodochrousPB1. Appl. Environ. Microbiol. 62: 749–755

    CAS  Google Scholar 

  • Tett VA, Willetts AJ & Lappin-Scott HM (1994) Enantioselective degradation of the herbicide mecoprop [2-(2-methyl-4-chlorophenoxy)propionic acid] by a mixed and pure bacterial culture. FEMS Microbiol. Ecol. 14: 191–200

    Article  CAS  Google Scholar 

  • Witschel M, Nagel S & Egli T (1997) Identification and characterization of the two-enzyme system catalyzing the oxidation of EDTA in the EDTA-degrading bacterial strain DSM 9103. J. Bacteriol. 179: 6937–6943

    CAS  Google Scholar 

  • Wolf K & Gilbert PA (1992) EDTA-Ethylenediaminetetraacetic acid. In: Hutzinger O (Ed) The Handbook of Environmental Chemistry, Vol 3 (pp 243–259). Springer, Berlin Heidelberg

    Google Scholar 

  • Xun L, Reeder RB, Plymale AE, Girvin DC & Bolton H (1996) Degradation of metal nitrilotriacetate complexes by nitrilotriacetate monooxygenase. Environ. Sci. Technol. 30: 1752–1755

    Article  CAS  Google Scholar 

  • Zipper C, Nickel K, Angst W & Kohler H-PE (1996) Complete microbial degradation of both enantiomers of the chiral herbicide mecopropo [(RS)-2-(4-chloro-2-methylphenoxy)propionic acid] in an enantioselective manner by Sphinogmonas herbicidivoranssp. nov. Appl. Environ. Microbiol. 62: 4318–4322

    CAS  Google Scholar 

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Witschel, M., Egli, T. Purification and characterization of a lyase from the EDTA-degrading bacterial strain DSM 9103 that catalyzes the splitting of [S,S]-ethylenediaminedisuccinate, a structural isomer of EDTA. Biodegradation 8, 419–428 (1997). https://doi.org/10.1023/A:1008267931018

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