Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Involvement of PEG-carboxylate dehydrogenase and glutathione S-transferase in PEG metabolism by Sphingopyxis macrogoltabida strain 103

  • 163 Accesses

  • 3 Citations


Sphingopyxis terrae and the Sphingopyxis macrogoltabida strains 103 and 203 are able to degrade polyethylene glycol (PEG). They possess the peg operon, which is responsible for the conversion of PEG to PEG-carboxylate-coenzyme A (CoA). The upstream (3.0 kb) and downstream (6.5 kb) regions of the operon in strain 103 were cloned and sequenced. The structure was well conserved between S. macrogoltabida strain 203 and S. terrae, except that two sets of transposases are absent in strain 203. The downstream region contains the genes for PEG-carboxylate dehydrogenase (PCDH), glutathione S-transferase (GST), tautomerase, and a hypothetical protein. The genes for pcdh and gst were transcribed constitutively and monocistronically, indicating that their transcription is independent of the operon regulation. PCDH and GST were expressed in Escherichia coli and characterized biochemically. PCDH is a homotetramer of 64-kDa subunits and contains one molecule of flavin adenine dinucleotide per subunit. The enzyme dehydrogenates PEG-carboxylate to yield glyoxylate, suggesting that the enzyme is the third enzyme involved in PEG degradation. GST is a homodimer of 28-kDa subunits. GST activity was noncompetitively inhibited by acyl-CoA and PEG-carboxylate-CoA, suggesting the interaction of GST with them. The proposed role for GST is to buffer the toxicity of PEG-carboxylate-CoA.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6


  1. Anraku Y, Heppel LA (1967) On the nature of the changes induced in Escherichia coli by osmotic shock. J Biol Chem 242:2561–2569

  2. Baker AL, Tolbert NE (1966) Glycolate oxidase (ferredoxin-containing form). Meth Enzymol 9:338–342

  3. Bergmeyer HU, Gowehn K, Grassl M (1974) In: Bergmeyer HU (ed) Enzymes as biochemical reagents. Academic, New York

  4. 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

  5. Charoenpanich J, Tani A, Moriwaki N, Kimbara K, Kawai F (2006) Dual regulation of a polyethylene glycol degradative operon by AraC-type and GalR-type regulators in Sphingopyxis macrogoltabida strain 103. Microbiology 152:3025–3034

  6. Enokibara S, Kawai F (1997) Purification and characterization of an ether bond-cleaving enzyme involved in the metabolism of polyethylene glycol. J Ferment Bioeng 83:549–554

  7. Everse J, Kaplan NO (1973) Lactate dehydrogenase: structure and function. Adv Enzymol Relat Areas Mol Biol 37:61–133

  8. Gamar Y, Gaunt JK (1971) Bacterial metabolism of 4-chloro-2-methylphenoxyacetate: formation of glyoxylate by side-chain cleavage. Biochem J 122:527–531

  9. Garden A, Levinthal C (1960) A fine-structure genetic and chemical study of the enzyme alkaline phosphatase of E. coli I. Purification and characterization of alkaline phosphatase. Biochim Biophys Acta 38:470–483

  10. Habig WH, Pabst MJ, Jakoby WB (1974) Glutathione S-transferase: the first step in mercapturic acid formation. J Biol Chem 249:7130

  11. Hawkins JM, Jones WE, Bonner FW, Gibson GG (1987) The effect of peroxisome proliferators on microsomal, peroxisomal, and mitochondrial enzyme activities in the liver and kidney. Drug Metab Rev 18:441–515

  12. Hu X, Fukutani A, Liu X, Kimbara K, Kawai F (2007) Isolation of bacteria able to grow on both polyethylene glycol (PEG) and polypropylene glycol (PPG) and their PEG/PPG dehydrogenases. Appl Microbiol Biotechnol 73:1407–1413

  13. Jakoby WB (1978) The glutathione S-transferase: a group of multi-functional detoxification proteins. Adv Enzymol 46:383–414

  14. Ji X, Zhang P, Armstrong RN, Gilliland GL (1992) The three-dimensional structure of a glutathione S-transferase from the Mu gene class. Structural analysis of the binary complex of isoenzyme 3–3 and glutathione at 2.2-Å resolution. Biochemistry 31:10169–10184

  15. Kawai F (2002) Microbial degradation of polyethers. Appl Microbiol Biotechnol 58:30–38

  16. Kawai F, Yamanaka H (1986) Biodegradation of polyethylene glycol by symbiotic mixed culture (obligate mutualism). Arch Microbiol 146:125–129

  17. Kawai F, Kimura T, Fukaya M, Tani Y, Ogata K, Ueno T, Fukami H (1978) Bacterial oxidation of polyethylene glycol. Appl Environ Microbiol 35:679–684

  18. Kawai F, Kimura T, Tani Y, Yamada H, Kurachi K (1985) Purification and characterization of polyethylene glycol dehydrogenase involved in the bacterial metabolism of polyethylene glycol. Appl Environ Microbiol 40:701–705

  19. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 227:680–685

  20. Mannervik B, Danielson UH (1988) Glutathione transferases: structure and catalytic activity. CRC Crit Rev Biochem 23:283–337

  21. Marmur JA (1961) A procedure for the isolation of deoxyribonucleic acid from microorganisms. J Mol Biol 3:208–218

  22. Masai E, Ichimura A, Sato Y, Miyauchi K, Katayama Y, Fukada M (2003) Roles of the enantioselective glutathione S-transferase in cleavage of β-aryl ether. J Bacteriol 185:1768–1775

  23. Miyamoto T, Yamamoto I (1994) Glutathione conjugates as the activated form of chalcones for glutathione S-transferase inhibition. J Pesticide Sci 19:53–58

  24. Nojiri H, Shintani M, Omori T (2004) Divergence of mobile genetic elements involved in the distribution of xenobiotic-catabolic capacity. Appl Microbiol Biotechnol 64:154–174

  25. Ohta T, Tani A, Kimbara K, Kawai F (2005) A novel nicotinoprotein aldehyde dehydrogenase involved in polyethylene glycol degradation. Appl Microbiol Biotechnol 68:639–646

  26. Ohta T, Kawabata T, Nishikawa K, Tani A, Kimbara K, Kawai F (2006) Analysis of amino acid residues involved in catalysis of polyethylene glycol dehydrogenase from Sphingopyxis terrae, using three-dimensional molecular modeling-based kinetic characterization of mutants. Appl Environ Microbiol 72:4388–4396

  27. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor

  28. Schilke B, Voisine C, Beinert H, Craig E (1999) Evidence for a conserved system for iron metabolism in the mitochondria of Saccharomyces cerevisiae. Proc Natl Acad Sci USA 96:10206–10211

  29. Shimizu S, Inoue K, Tani Y, Yamada H (1979) Enzymatic microdetermination of serum free fatty acids. Anal Biochem 98:341–345

  30. Silva C, Loyola G, Valenzuela R, Garcia-Huidobro T, Monasterio O, Bronfman M (1999) High-affinity binding of fatty acyl-CoAs and peroxisome proliferator-CoA esters to glutathione S-transferases: effect on enzymatic activity. Eur J Biochem 266:143–150

  31. Sugimoto M, Tanabe M, Hataya M, Enokibara S, Duine JA, Kawai F (2001) The first step in polyethylene glycol degradation by Sphingomonads proceeds via a flavoprotein alcohol dehydrogenase containing flavin adenine dinucleotide. J Bacteriol 183:6694–6698

  32. Takeuchi M, Kawai F, Shimada Y, Yokota A (1993) Taxonomic study of polyethylene glycol-utilizing bacteria: emended description of the genus Sphingomonas and new descriptions of Sphingomonas macrogoltabidus sp. nov., Sphingomonas sanguis sp. nov. and Sphingomonas terrae sp. nov. System Appl Microbiol 16:227–238

  33. Takeuchi M, Hamana K, Hiraishi A (2001) Proposal of the genus Sphingomonas sensu stricto and three new genera, Sphingobium, Novosphingobium and Sphingopyxis, on the basis of phylogenetic and chemotaxonomic analyses. Int J Syst Evol Microbiol 51:1405–1417

  34. Tan HM (1999) Bacterial catabolic transposons. Appl Microbiol Biotechnol 51:1–12

  35. Tani A, Charoenpanich J, Mori T, Takeichi M, Kimbara K, Kawai F (2007) Structure and conservation of a polyethylene glycol-degradative operon in sphingomonads. Microbiology 153:338–346

  36. Tani A, Somyoonsap P, Minami T, Kimbara K, Kawai F (2008) Polyethylene glycol (PEG)-carboxylate-CoA synthetase is involved in PEG metabolism in Sphingopyxis macrogoltabida strain 103. Arch Microbiol 189:407–410

  37. Widersten M, Holmstrom E, Mannervik B (1991) Cysteine residues are not essential for the catalytic activity of human class Mu glutathione transferase M1a-1a. FEBS Lett 293:156–159

  38. Wilce MCJ, Parker MW (1994) Structure and function of glutathione S-transferases. Biochim Biophys Acta 1205:1–18

  39. Yamanaka H, Kawai F (1989) Purification and characterization of constitutive polyethylene glycol (PEG) dehydrogenase of a PEG 4000-utilizing Flavobacterium sp. No. 203. J Ferment Bioeng 67:324–330

  40. Yamashita M, Tani A, Kawai F (2005) A new ether bond-splitting enzyme found in Gram-positive polyethylene glycol 6000-utilizing bacterium, Pseudonocardia sp. strain K1. Appl Microbiol Biotechnol 66:174–179

Download references


This work was partly supported by the Wesco Scientific Promotion Foundation and Grant-in-Aid for Scientific Research (C) 14560068 to FK and Grant-in-Aid for Young Scientists (B) 1919780060 to AT from the Japan Society for the Promotion of Science. This work was linked with the JSPS-NRCT Core University Program.

Author information

Correspondence to Fusako Kawai.

Additional information

American Journal Experts (http://www.jounalexperts.com) edited the manuscript.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Somyoonsap, P., Tani, A., Charoenpanich, J. et al. Involvement of PEG-carboxylate dehydrogenase and glutathione S-transferase in PEG metabolism by Sphingopyxis macrogoltabida strain 103. Appl Microbiol Biotechnol 81, 473–484 (2008). https://doi.org/10.1007/s00253-008-1635-7

Download citation


  • Polyethylene glycol (PEG)
  • peg operon
  • PEG-carboxylate dehydrogenase
  • Glutathione S-transferase
  • Sphingopyxis macrogoltabida