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Enhancing survival of Escherichia coli by expression of azoreductase AZR possessing quinone reductase activity

  • Guangfei Liu
  • Jiti Zhou
  • Ruofei Jin
  • Mi Zhou
  • Jing WangEmail author
  • Hong Lu
  • Yuanyuan Qu
Biotechnologically Relevant Enzymes and Proteins

Abstract

Quinone reductase activity of azoreductase AZR from Rhodobacter sphaeroides was reported. High homologies were found in the cofactor/substrate-binding regions of quinone reductases from different domains. 3D structure comparison revealed that AZR shared a common overall topology with mammal NAD(P)H/quinone oxidoreductase NQO1. With menadione as substrate, the optimal pH value and temperature were pH 8–9 and 50°C, respectively. Following the ping-pong kinetics, AZR transferred two electrons from NADPH to quinone substrate. It could reduce naphthoquinones and anthraquinones, such as menadione, lawsone, anthraquinone-2-sulfonate, and anthraquinone-2,6-disulfonate. However, no activity was detected with 1,4-benzoquinone. Dicoumarol competitively inhibited AZR’s quinone reductase activity with respect to NADPH, with an obtained Ki value of 87.6 μM. Significantly higher survival rates were obtained in Escherichia coli YB overexpressing AZR than in the control strain when treated by heat shock and oxidative stressors such as H2O2 and menadione.

Keywords

Quinone reductase Azoreductase Oxidative stress Heat shock Survival 

References

  1. Andrade SLA, Patridge EV, Ferry JG, Einsle O (2007) Crystal structure of the NADH:quinone oxidoreductase WrbA from Escherichia coli. J Bacteriol 189:9101–9107CrossRefGoogle Scholar
  2. Asher G, Dym O, Tsvetkov P, Adler J, Shaul Y (2006) The crystal structure of NAD(P)H quinone oxidoreductase 1 in complex with its potent inhibitor dicoumarol. Biochemistry 45:6372–6378CrossRefGoogle Scholar
  3. Baker CJ, O’Neill NR, Keppler LD, Orlandi EW (1991) Early responses during plant-bacteria interactions in tobacco cell suspensions. Phytopathology 81:1504–1507CrossRefGoogle Scholar
  4. Beyer RE (1994) The relative essentiality of the antioxidative function of coenzyme Q-the interactive role of DT-diaphorase. Mol Aspects Med 15(suppl.):117–129CrossRefGoogle Scholar
  5. Bianchet MA, Faig M, Amzel LM (2004) Structure and mechanism of NAD(P)H:quinone acceptor oxidoreductases (NQO). Methods Enzymol 382:144–174CrossRefGoogle Scholar
  6. Blümel S, Knackmuss HJ, Stolz A (2002) Molecular cloning and characterization of the gene coding for the aerobic azoreductase from Xenophilus azovorans KF46F. Appl Environ Microbiol 68:3948–3955CrossRefGoogle Scholar
  7. 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 72:248–254Google Scholar
  8. Cases I, de Lorenze V (2005) Genetically modified organisms for the environment: stories of success and failure and what we have learned from them. Int Microbiol 8:213–222PubMedGoogle Scholar
  9. Cenas N, Nemeikaite-Ceniene A, Sergediene E, Nivinskas H, Anusevicius Z, Sarlauskas J (2001) Quantitative structure-activity relationships in enzymatic single-electron reduction of nitroaromatic explosives: implications for their cytotoxicity. Biochim Biophys Acta 1528:31–38CrossRefGoogle Scholar
  10. Chen H (2006) Recent advances in azo dye degrading enzyme research. Curr Protein Pept Sci 7:101–111CrossRefGoogle Scholar
  11. Chen S, Wu K, Zhang D, Sherman M, Knox R, Yang CS (1999) Molecular characterization of binding of substrates and inhibitors to DT-diaphorase: combined approach involving site-directed mutagenesis, inhibitor-binding analysis, and computer modeling. Mol Pharmacol 56:272–278CrossRefGoogle Scholar
  12. Deller S, Sollner S, Trenker-El-Toukhy R, Jelesarov I, Gubitz GM, Macheroux P (2006) Characterization of a thermostable NADPH:FMN oxidoreductase from the mesophilic bacterium Bacillus subtilis. Biochemistry 45:7083–7091CrossRefGoogle Scholar
  13. Deller S, Macheroux P, Sollner S (2008) Flavin-dependent quinone reductases. Cell Mol Life Sci 65:141–160CrossRefGoogle Scholar
  14. Dos Santos AB, Cervantes FJ, Van Lier JB (2007) Review paper on current technologies for decolourisation of textile wastewaters: pespectives for anaerobic biotechnology. Biores Technol 98:2369–2385CrossRefGoogle Scholar
  15. Ercal N, Gurer-Orhan H, Aykin-Burns N (2001) Toxic metals and oxidative stress part I: mechanisms involved in metal-induced oxidative damage. Curr Top Med Chem 1:529–539CrossRefGoogle Scholar
  16. Garbisu C, Alkorta I (1999) Utilization of genetically engineered microorganisms (GEMs) for bioremediation. J Chem Technol Biotechnol 74:599–606CrossRefGoogle Scholar
  17. Gonzalez CF, Ackerley DF, Lynch SV, Matin A (2005) ChrR, a soluble quinone reductase of Pseudomonas putida that defends against H2O2. J Biol Chem 280:22590–22595CrossRefGoogle Scholar
  18. Hong Y, Wang G, Maier RJ (2008) The NADPH quinone reductase MdaB confers oxidative stress resistance to Helicobacter hepaticus. Microb Pathog 44:169–174CrossRefGoogle Scholar
  19. Iyanagi T (1987) On the mechanisms of one- and two-electron transfer by flavin enzymes. Chemica Scripta 27A:31–36Google Scholar
  20. Kim HJ, Kang BS, Park JW (2005) Cellular defense against heat shock-induced oxidative damage by mitochondrial NADP+-dependent isocitrate dehydrogenase. Free Radic Res 39:441–448CrossRefGoogle Scholar
  21. Laskowski MJ, Dreher KA, Gehring MA, Abel S, Gensler AL, Sussex IM (2002) FQR1, a novel primary auxin-response gene, encodes a flavin mononucleotide-binding quinone reductase. Plant Physiol 128:578–590CrossRefGoogle Scholar
  22. Li R, Bianchet MA, Talalay P, Amzel LM (1995) The three-dimensional structure of NAD(P)H:quinone reductase, a flavoprotein involved in cancer chemoprotection and chemotherapy: mechanism of the two-electron reduction. Proc Natl Acad Sci U S A 92:8846–8850CrossRefGoogle Scholar
  23. Liger D, Graille M, Zhou CZ, Leulliot N, Quevillon-Cheruel S, Blondeau K, Janin J, van Tilbeurgh H (2004) Crystal structure and functional characterization of yeast YLR011wp, an enzyme with NAD(P)H-FMN and ferric iron reductase activities. J Biol Chem 279:34890–34897CrossRefGoogle Scholar
  24. Lind C, Hochstein P, Ernster L (1982) DT-diaphorase as a quinone reductase: a cellular control device against semiquinone and superoxide radical formation. Arch Biochem Biophys 216:178–185CrossRefGoogle Scholar
  25. Liu G, Zhou J, Lv H, Xiang X, Wang J, Zhou M, Qv Y (2007) Azoreductase from Rhodobacter sphaeroides AS1.1737 is a flavodoxin that also functions as nitroreductase and flavin mononucleotide reductase. Appl Microbiol Biotechnol 76:1271–1279CrossRefGoogle Scholar
  26. Nicholas KB, Nicholas HB, Jr, Deerfield DW (1997) GeneDoc: analysis and visualization of genetic variation. EMBnet News 4:1–4Google Scholar
  27. Paterson ES, Boucher SE, Lambert IB (2002) Regulation of the nfsA gene in Escherichia coli by SoxS. J Bacteriol 184:51–58CrossRefGoogle Scholar
  28. Patridge EV, Ferry JG (2006) WrbA from Escherichia coli and Archaeoglobus fulgidus is an NAD(P)H:quinone oxidoreductase. J Bacteriol 188:3498–3506CrossRefGoogle Scholar
  29. Prestera T, Prochaska HJ, Talalay P (1992) Inhibition of NAD(P)H:(quinone-acceptor) oxidoreductase by cibacron blue and related anthraquinone dyes: a structure-activity study. Biochemistry 31:824–833CrossRefGoogle Scholar
  30. Ross D, Siegel D (2004) NAD(P)H:quinone oxidoreductase 1 (NQO1, DT-diaphorase), functions and pharmacogenetics. Methods Enzymol 382:115–144CrossRefGoogle Scholar
  31. Russ R, Rau J, Stolz A (2000) The function of cytoplasmic flavin reductases in the reduction of azo dyes by bacteria. Appl Environ Microbiol 66:1429–1434CrossRefGoogle Scholar
  32. Soballe B, Poole RK (1999) Microbial ubiquinones: multiple roles in respiration, gene regulation and oxidative stress management. Microbiology 145:1817–1830CrossRefGoogle Scholar
  33. Soballe B, Poole RK (2000) Ubiquinone limits oxidative stress in Escherichia coli. Microbiology 146:787–796CrossRefGoogle Scholar
  34. Sollner S, Nebauer R, Ehammer H, Prem A, Deller S, Palfey BA, Daum G, Macheroux P (2007) Lot6p from Saccharomyces cerevisiae is a FMN-dependent reductase with a potential role in quinone detoxification. FEBS J 274:1328–1339CrossRefGoogle Scholar
  35. Stolz A (2001) Basic and applied aspects in the microbial degradation of azo dyes. Appl Microbiol Biotechnol 56:69–80Google Scholar
  36. Wang G, Maier RJ (2004) An NADPH quinone reductase of Helicobacter pylori plays an important role in oxidative stress resistance and host colonization. Infect Immun 72:1391–1396CrossRefGoogle Scholar
  37. Yan B, Zhou J, Wang J, Du C, Hou H, Song Z, Bao Y (2004) Expression and characteristics of the gene encoding azoreductase from Rhodobacter sphaeroides AS1.1737. FEMS Microbiol Lett 236:129–136CrossRefGoogle Scholar
  38. Zenno S, Koike H, Kumar AN, Jayaraman R, Tanokura M, Saigo K (1996) Biochemical characterization of NfsA, the Escherichia coli major nitroreductase exhibiting a high amino acid sequence homology to Frp, a Vibrio harveyi flavin oxidoreductase. J Bacteriol 178:4508–4514CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Guangfei Liu
    • 1
  • Jiti Zhou
    • 1
  • Ruofei Jin
    • 1
  • Mi Zhou
    • 1
  • Jing Wang
    • 1
    Email author
  • Hong Lu
    • 1
  • Yuanyuan Qu
    • 1
  1. 1.School of Environmental and Biological Science and TechnologyDalian University of TechnologyDalianPeople’s Republic of China

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