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The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Molecular Characterization of Epoxide Hydrolase from Aspergillus niger LK using Phylogenetic Analysis

진화적 유연관계 분석을 통한 Aspergillus niger LK의 Epoxide Hydrolase의 특성분석

  • 김희숙 (경성대학교 공과대학 식품공학과) ;
  • 이은열 (경성대학교 공과대학 식품공학과) ;
  • 이수정 (경성대학교 공과대학 식품공학과) ;
  • 이지원 (경성대학교 공과대학 식품공학과)
  • Published : 2004.02.01
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Abstract

A gene coding for epoxide hydrolase (EH) of Aspergillus niger LK, a fungus possessing the enantioselective hydrolysis activity for racemic epoxides, was characterized by phylogenetic analysis. The deduced protein of A. niger LK epoxide hydrolase shares significant sequence similarity with several bacterial EHs and mammalian microsomal EHs (mEH) and belongs to the a/${\beta}$ hydrolase fold family. EH from A. niger LK had 90.6% identity with 3D crystal structure of lqo7 in Protein Data Bank. Sequence comparison with other source EHs suggested that Asp$\^$l92/, Asp$\^$374/ and His$\^$374/ constituted the catalytic triad. Based on the multiple sequence comparison of the functional and structural domain sequence, the phylogenetic tree between relevant epoxide hydrolases from various species were reconstructed by using Neighbor-Joining method. Genetic distances were so far as 1.841-2.682 but characteristic oxyanion hole and catalytic triad were highly conserved, which means they have diverged from a common ancestor.

Racemic epoxide에 대한 입체선택적 가수분해능을 가지고 있는 곰팡이, Aspergillus niger LK로부터 epoxide hydrolase (EH, EC 3.3.2.3) 유전자의 진화적 유연관계 분석을 행하였다. A. niger LK의 EH 염기서열로부터 유추한 EH 단백질 아미노산 서열은 여러 박테리아의 EH들 및 포유동물의 microsomal EH들과 유의적인 유사성을 가지고 있었으며 a/$\beta$ hydrolase fold family에 속하였다. A. niger LK의 EH 단백질의 입체구조예측은 Protein Data Bank에 수록된 lqo7의 3D 결정구조와 90.6% identity를 가지는 것으로 나타났으며 다른 EH들의 아미노산 서열비교를 행한 결과 Asp$^{192}$ , Asp$^{348}$ 및 His$^{374}$ 이 catalytic triad를 구성하고 있는 것으로 추정되었다. 여러 생물종의 EH서열을 기능적 및 구조적 domain 서열을 기초로 하여 multiple sequence alignment를 행하고 Neighbor-Joining/UPGMA method를 이용하여 계통수를 복원한 결과 다른 생물종들의 EH와의 진화거리는 서로 1.841∼2.682로 멀었으나 EH의 기능을 가지기 위한 oxyanion hole 및 a/$\beta$ hydrolase fold family의 catalytic triad는 잘 보존되고 있어 공통조상으로부터 진화되어 왔음을 알 수 있었다.

Keywords

References

  1. Curr. Opin. Chem. Biol. v.5 Synthetic applications of epoxide hydrolases Archelas,A.;R.Furstoss https://doi.org/10.1016/S1367-5931(00)00179-4
  2. Curr. Opin biotechnol v.12 Microbial epoxide hydrolases for preparative biotransformations Steinreiber,A.;K.Faber https://doi.org/10.1016/S0958-1669(01)00262-2
  3. J. Mol. Catal. B: Enz. v.6 Epoxide hydrolases from yeasts and other sources: versatile tools in biocatalysis Weijers,C.A.G.M.;J.A.M. de Bont https://doi.org/10.1016/S1381-1177(98)00123-4
  4. Oral Oncol. v.39 Epoxide hydrolase genotype and oralaryngeal cancer risk: interaction with GSTM1 genotype Park,J.Y.;S.P.Schantz;P.Lazarus https://doi.org/10.1016/S1368-8375(03)00008-3
  5. Appl. Environ. Microbiol. v.65 Isolation and characterization of the epoxide hydrolase encoding gene from Xanthophyllimyces dendrorhous Visser,H.;J.A.M. de Bont;J.C.Verdoes
  6. Kor. J. Biotechnol. Bioeng. v.17 Epoxide hydrolase-catalyzed hydrolytic biological synthesis of chiral epoxides Lee,E.Y.
  7. Tetrahedron v.50 Chemical and biological synthesis of chiral epoxides Besse,P.;H.Veschambre https://doi.org/10.1016/S0040-4020(01)85362-X
  8. Curr. Opin. biotechnol. v.14 Biocatalytic conversion of epoxides de Vries, E.J.;D.B.Janssen https://doi.org/10.1016/S0958-1669(02)00013-7
  9. Biochem. J. v.344 Clonin and molecular characterization of a soluble epoxide hydrolase from Aspergillus niger that is related to mammalian microsomal epoxide hydrolase Arand,M.;H.Hemmer;H.Durk;J.Baratti;A.Archelas;R.Furstoss;F.Oesch https://doi.org/10.1042/0264-6021:3440273
  10. J. Am. Chem. Soc. v.121 Mutation of tyrosine residues involved in the alkylation half reaction of epoxide hydrolase from Agrobacterium radiobacter AD1 results in improved enantioselectivity Rink,R.;J.H.L.Spelberg;R.J.Pieters;J.Kingma;M.Nardini;R.M.Kellogg;B.W.Dijkstra;D.B.Janssen https://doi.org/10.1021/ja990501o
  11. Kor. J. Biotechnol. Bioeng. v.16 Cloning and molecular characterization of epoxide hydrolase from Aspergillus niger LK Lee,E.Y.;H.S.Kim
  13. Protein, Nucleic acid Enzyme v.46 Knowing similarity in protein 3D structure Kidera,A.
  15. Proteins v.9 Database of homologyderived structures and the structural meaning of sequence alignment Sander,C.;R.Schneider https://doi.org/10.1002/prot.340090107
  19. FEBS Lett. v.338 Sequence similarity of mammalian epoxide hydrolases to the bacterial haloalkane dehalogenase and other related proteins. Implication for the potential catalytic mechanism of enzymatic epoxide hydrolysis Arand,M.;D.F.Grant;J.K.Beetham;T.Friedberg;F.Oesch;B.D.hammock https://doi.org/10.1016/0014-5793(94)80278-5
  20. Proc. Natl. Acad. Sci. USA v.84 Molecular cloning and amino acid sequence of leukotriene A₄hydrolase Funk,C.D.;O.Radmark;J.Y.Fu;T.Matsumoto;H.Jornvall;T.Shimizu;B.Samuelsson https://doi.org/10.1073/pnas.84.19.6677
  21. J. Biol. Chem. v.256 The hepatic microsomal biotransformation of △5-steroids to 5α, 6β-glycols via α- and β-dpoxides Watabe,T.;M.Kanai;M.Isobe;N.Ozawa
  22. J. Bacteriol. v.180 Limonene-1,2-epoxide to a novel class of epoxide hydrolases van der Werf, M.J.;K.M.Overkamp;J.A. de Bont
  23. J. Biol. Chem. v.272 Primary structure and catalytic mechanism of the epoxide hydrolase from Agrobaccerium radiobacter ADSI Rink,R.;M.Fennema;M.Smids;U.Dehmel;D.B.Janssen https://doi.org/10.1074/jbc.272.23.14650
  24. Eur. J. Biochem. v.267 Purification, molecular cloning and ethylene-inducible expression of a soluble-type epoxide hydrolase from soybean Arahira,M.;V.H.Nong;K.Udaka;C.Fukazawa https://doi.org/10.1046/j.1432-1327.2000.01276.x
  25. Arch. Biochem. Biophys. v.305 cDNA cloning and expression of a soluble epoxide hydrolase from human liver Beetham,J.K.;T.Tian;B.D.Hammock https://doi.org/10.1006/abbi.1993.1411
  26. J. Bacteriol. v.183 Genome sequence and comparative analysis of the solvent-producing bacerium Clostridium acetobutylicum Nolling,J.;G.Breton;M.V.Omelchenko;K.S.Markarova;Q.Zeng;R.Gibson;H.M.Lee;J.Dubois;D.Qiu;J.Hitti;Y.I.Wolf;R.L.Tatusov;F.Sabathe;L.Doucette-stamm;P.Soucaille;M.J.Daly;G.N.Bennett;E.V.Koomin;D.R.Smith https://doi.org/10.1128/JB.183.16.4823-4838.2001
  27. Putative epoxide hydrolase. Direct submitted to the EMBL/GenBank/DDBJ database (entry AAL17927.1) Derbyshire,K.M.;L.M.Parsons;J.DeVost
  28. Science v.287 The genome sequence of Drosophila melanogaster Adams,M.D.;S.E.Celniker;R.A.Holt;C.A.Evans;J.D.Gocayne;P.G.Amanatides;S.E.Scherer;P.W.Li;R.A.Hoskins;R.F.Galle https://doi.org/10.1126/science.287.5461.2185
  29. J. Biol. Chem. v.263 Human microsoml xenobiotic epoxide hydrolase. Complementary DNA sequence, complementary DNA-directed expression in COS-1 cells, and chromosomal localization Skoda,R.C.;A.Demierre;O.W.McBride;F.J.Gonzalez;U.A.Meyer