Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Identification of Genes Involved in Decolorization of Crystal Violet and Malachite Green in Citrobacter sp.

Citrobacter sp.에서 crystal violet와 malachite green 색소분해에 관여하는 유전자들의 동정

  • Lee, Young-Mi (College of Natural Resources and Life Science, Dong-A University) ;
  • Jang, Moon-Sun (College of Natural Resources and Life Science, Dong-A University) ;
  • Kim, Seok-Jo (College of Natural Resources and Life Science, Dong-A University) ;
  • Park, Yong-Lark (College of Natural Resources and Life Science, Dong-A University) ;
  • Cho, Young-Su (College of Natural Resources and Life Science, Dong-A University) ;
  • Lee, Young-Choon (College of Natural Resources and Life Science, Dong-A University)
  • Published : 2004.02.01
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Abstract

To identify genes involved in the decolorization of both crystal violet and malachite green, we isolated random mutants generated by transposon insertion in triphenylmethane-decolorizing bacterium, Citrobacter sp. The resulting mutant bank yielded 14 mutants with complete defect in color removal capability of both crystal violet and malachite green. Southern hybridization with a Tn5 fragment as a probe showed a single hybridized band in 5 mutants and these mutants appeared to have insertions at different sites of the chromosome. Tn5-inserted genes were isolated and the DNA sequence flanking Tn5 was determined. From comparison with a sequence database, putative protein products encoded by cmg genes were identified as follows. cmg 2 is MaIC protein in maltose transport system; cmg 6 is transcriptional regulator (LysR-type): cmg 12 is a putative oxidoreductase. The sequences deduced from two cmg genes, cmg 8 and cmg 11, showed no significant similarity to any protein with a known function. Therefore, these results indicate that these two cmg genes encode unidentified proteins responsible for decolorization of both crystal violet and malachite green.

Crystal violet와 malachite green색소 분해에 관여하는 유전자들을 규명하기 위하여 색소분해능을 가진 Citrobacter sp.의 염색체 DNA속에의 transposon 도입에 의해 생성된 무작위 변이주들이 분리되었다. 이들 변이 주들로부터 두가지 색소분해능을 소실한 14개의 변이주들이 선별되었고, 이들로부터 염색체 DNA를 분리하여 EcoRl으로 절단한 후 Tn5 단편을 probe로하여 Southern hybridization을 행한 결과, 염색체 DNA상의 각각 다른 부위에 Transposon이 Single 삽입된 5개의 변이주 (Cmg2, Cmg6, Cmg8, Cmgll, Cmg12)가 최종적으로 분리되었다. 이들 변이주들의 Transposon 삽입부위 주위의 염기서열과 이로부터 유추되는 아미노산서열을 database상에 등록되어 있는 유전자의 염기서열과 단백질의 아미노산 서열에 대한 상동성을 비교한 결과, Cmg2는 대장균 maltose trnasporter (Mal C)이고, Cmg6은 LysR-type 전사조절 단백질이며, Cmg12는 산화환원효소를 코드하는 유전자인 것으로 알려졌고, 나머지 Cmg8과 Cmg11은 아직까지 기능이 알려져 있지 않은 단백질을 코드하는 유전자인 것으로임이 판명되었다.

Keywords

References

  1. J. Fish. Dis. v.8 Malachite green: a review Alderman.D.J. https://doi.org/10.1111/j.1365-2761.1985.tb00945.x
  2. Enzyme Microb. Technol. Microbial decolorization of triphenylmethane and azo dyes by Citrobacter sp. An.S.Y.;K.W.Kim;Y.L.Choi;Y.S.Cho;S.K.Min;I.H.Cha;C.H.Kim;Y.C.Lee
  3. vitro. Mutat. Res. v.58 Cytogenetic toxicity of gentian violet and crystal violet on mammalian cells Au,W.;S.Pathak;C.J.Collie;T.C.Hsu https://doi.org/10.1016/0165-1218(78)90019-8
  4. Mutat. Res. v.66 Further study of the genetic toxicology of gentian violet Au,W.;M.A.Butler;S.F.Bloom;T.S.Matmey https://doi.org/10.1016/0165-1218(79)90054-5
  5. Enzyme Microb. Technol. v.22 Biodegradation of triphenylmethane dyes Azmi,W.;R.K.Sani;U.C.Banerjee https://doi.org/10.1016/S0141-0229(97)00159-2
  6. Biochem. J. v.17 The inhibition of 1,1,1-trichloro-2,2-bis-(p-chlorophenyl)ethane (DDT) dehydrochlorinase and glutathione S-aryltransferase in grassgrub and housefly preparations Balabaskaran,S.;J.N.Smith
  7. Biores. Technol. v.58 Microbial decolorization of textile-dye-containing effluents: a review Banat,I.M.;P.Nigam;C.Singh;R.Marchant https://doi.org/10.1016/S0960-8524(96)00113-7
  8. Science v.277 The complete genome seqyence of Escherichia coli K-12 Blattner,F.R.;G.PlunkettⅢ;C.A.Bloch;N.T.Perna;V.Burland;M.Riley;J.Collado-Vides;J.D.Glasner;C.K.Rode;G.F.Mayhew;J.Gregor;N.M.Davis;H.A.Kirkpatrick;M.A.Goeden;D.J.Rose;B.Mau;Y.Shao https://doi.org/10.1126/science.277.5331.1453
  9. Microbiol. Mol. Biol. Rev. v.62 Maltose/maltodextrin ssystem of Escherichia coli: transport, metabolism, and regulation Boos,W.;H.Shuman
  10. Appl. Environ. Microbiol. v.54 Biodegradation of crystal violet by the white rot fungus Phanerochaete chrysosporium Bumpus,J.A.;B.J.Brock
  11. Arch. Biochem. Biophys. v.107 Metabolism of triphenylmethane and triphenylcabinol Cornish,H.H.;E.Zamora;R.E.Bahor https://doi.org/10.1016/0003-9861(64)90337-6
  12. J. Am. Coll. Toxicol. v.15 Malachite green: a toxicological review Culp,S.J.;F.A.Beland https://doi.org/10.3109/10915819609008715
  13. Mol. Microbiol. v.7 Membrane topology of MalG, an inner membrane protein from the maltose tramsport system of Escherichia coli. Dassa,E.;S.Muir https://doi.org/10.1111/j.1365-2958.1993.tb01094.x
  14. Biochem. Pharmacol. v.45 Inhibition of glutathione S-transferases from rat liver by basic triphenylmethane dyes Debnam,P.;S.Glanville;A.Clark https://doi.org/10.1016/0006-2952(93)90274-Z
  15. Mol. Microbiol. v.19 Regulatory noise in prokaryotic promoter: how bacteria learn to respond to novel environmental signals de Lorenzo,V.;J. Perez-Martin https://doi.org/10.1111/j.1365-2958.1996.tb02463.x
  16. J. Appl. Toxicol. v.19 Mutagenicity of malachite green and leucomalachite green in Fessard,V.;T.Godard,;S.Huet;A.Mourot;J.M.Poul https://doi.org/10.1002/(SICI)1099-1263(199911/12)19:6<421::AID-JAT595>3.0.CO;2-6
  17. Cur. Opin. Biotechnol. v.11 Bacterial pollutants triggering biodegradaion of aromatic pollutants Diaz,E.;M.A.Prieto https://doi.org/10.1016/S0958-1669(00)00126-9
  18. Encyclopedia of Chemical Technology v.8 Dyes and dyes intermediates Gregory,P.;J.L.Kroschwitz(ed.)
  19. Mol. Pharmacol. v.22 Microsomal reduction of gentian violet. Evidence for cytochrome P-450 catalyzed free radical formation Harrelson,W.G.;R.P.Mason
  20. Appl. Environ. Microbiol. v.63 Reduction of malachite green to leucomalachite green by intestinal bacteria Henderson,A.L.;T.C.Schmitt;T.M.Heinze;C.E.Cerniglia
  21. J. Pharm. Sci. v.89 Effect of molecular structure on the performance of triarylemthane dyes as therapeutic agents for photochemical Purging of autologous bone marrow grafts from residual tumor cells Indig,G.L.;G.S.Anderson;M.G.Nichols;J.A.Bartlett;W.S.Mellon;F.Sieber https://doi.org/10.1002/(SICI)1520-6017(200001)89:1<88::AID-JPS9>3.0.CO;2-K
  22. Antimicrob. Agents Chemother. v.47 Decolorization of malachite green and crystal violet by waterborne pathogenic mycobacteria Jone,J.J.;J.O.Falkinham https://doi.org/10.1128/AAC.47.7.2323-2326.2003
  23. Drugs of choice Drugs for intestinal parasitism Kean,B.H.;D.W.Haskins;W.Modell(ed.)
  24. Poult. Sci. v.55 A study of the feasibility of the use of gentian violet as a fungistat for poultry feed Kingsland,G.V.;J.Anderson https://doi.org/10.3382/ps.0550852
  25. J. Mol. Biol. v.3 A procedure for the isolation of deoxyribonucleic acid from microorganisms Marmur,J. https://doi.org/10.1016/S0022-2836(61)80047-8
  26. Environ. Sci. Technol. v.14 Aromatic amines in and near the Buffalo River Nelson,C.R.;R.A. Hites https://doi.org/10.1021/es60169a020
  27. Toxicol. Lett. v.81 Inhibition of DNA synthesis in primary rat hepatocyte cultures by malachite green: a new liver tumor promoter Rao,K.V.K. https://doi.org/10.1016/0378-4274(95)03413-7
  28. Molecular cloning: laboratory manual(2nd edn.) Sambrook,J.;E.F.Fritsch;T.Maniatis
  29. Gene v.56 A series of Tn5 variants with various drug-resistance markers and suicide vector for transposon mutagenesis Sasakawa,C.;M.Yoshikawa https://doi.org/10.1016/0378-1119(87)90145-4
  30. Annu. Rev. Microbiol. v.47 Molecular biology of the LysR family of transcriptional regulators Schell,M.A. https://doi.org/10.1146/annurev.mi.47.100193.003121
  31. Biotechnol. Lett. v.20 Decolorization of artificial dyes by peroxidase from the white-rot fungus pleurotus ostreatus Shin,K.S.;C.J.Kim https://doi.org/10.1023/A:1005301812253
  32. Curr. Microbiol. v.30 Decolorization of triphenylmethane dyse by the bids nest fungus Cyathu bukkeri Vasdev,K.;R.C.Kuhad;R.K.Saxena https://doi.org/10.1007/BF00295500