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Cloning and expression of a lipoxygenase from Pseudomonas aeruginosa 42A2

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Abstract

In order to produce (S) 10-monohydroxy-8E-octadecenoic acid (MHOD) from oleic acid, a full-length probable lipoxygenase cDNA from Pseudomonas aeruginosa 42A2 was cloned and expressed in Escherichia coli BL21(DE3). The recombinant protein was purified by affinity chromatography to electrophoretic homogeneity and specifically stained. Its molecular mass was 70 kDa. The activity of the rec-LOX with oleic acid was about 30% of that of the prefered substrate, linoleic acid (100%). Bacterial LOX forms a new subfamily in the lipoxygenase phylogenetic tree.

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Abbreviations

IPTG:

(isopropyl β-D-thiogalactoside)

MHOD:

(S)10-monohydroxy-8E-octadecenoic acid

MHOOD:

(S)10-monohydroperoxy-8E-octadecenoic

PAGE:

polyacrylamide gel electrophoresis

PCR:

polymerase chain reaction

PMSF:

phenylmethyl-sulphonyl fluoride

TCP:

total cell protein X-Gal(5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside)

References

  • S.F. Altschul T.L. Madden A.A. Schaffer J. Zhang Z. Zhang W. Miller D.J. Lipman (1997) ArticleTitleGapped BLAST and PSI-BLAST: a new generation of protein database search programs Nucleic Acids Res. 25 3389–3402 Occurrence Handle10.1093/nar/25.17.3389 Occurrence Handle1:CAS:528:DyaK2sXlvFyhu7w%3D Occurrence Handle9254694

    Article  CAS  PubMed  Google Scholar 

  • E. André K. Hou (1932) ArticleTitlesur la présence d’une oxydase des lipides ou lipoxydase dans la graine de sojaGlycine soja Lieb Comptes Rendus 194 645–647

    Google Scholar 

  • E. Bleé A.L. Wilcox L.J. Marnett F. Schuber (1993) ArticleTitleMechanism of reaction of fatty acid hydroperoxides with soybean peroxygenase J. Biol. Chem. 268 1708–1715 Occurrence Handle8420948

    PubMed  Google Scholar 

  • A.R. Brash (1999) ArticleTitleLipoxygenases. Occurrencefunctions, catalysis, and acquisition of substrate J. Biol. Chem. 274 23679–23682 Occurrence Handle10446122 Occurrence Handle1:CAS:528:DyaK1MXlsVOnsrc%3D

    PubMed  CAS  Google Scholar 

  • M. Busquets V. Deroncele J. Vidal-Mas E. Rodriguez A. Guerrero A. Manresa (2004) ArticleTitleIsolation and characterization of a lipoxygenase from Pseudomonas 42A2 responsible of the biotransformation of oleic acid into (S)-(E)-10-hydroxy-8-octadecenoic acid Antonie van Leeuvenhoek 85 129–139 Occurrence Handle1:CAS:528:DC%2BD2cXitFCmtrY%3D

    CAS  Google Scholar 

  • A. Boer Particlede C. Smidth-Dannert (2003) ArticleTitleRecent effort in engineering microbial cells to produce new chemical compounds Curr. Opin. Biol. Chem. 7 273–278

    Google Scholar 

  • A. Guerrero I. Casals M. Busquets Y. León A. Manresa (1997) ArticleTitleOxidation of oleic acid to (E)-10-hydroperoxy-8-octadecenoic acid and (E)-10-hydroxy-8-ocatadecenoic acids by Pseudomonas sp. 42A2 Biochimica et Biophysica Acta 1347 75–81 Occurrence Handle9233689 Occurrence Handle1:CAS:528:DyaK2sXkt1Sjtr0%3D

    PubMed  CAS  Google Scholar 

  • M. Hamberg B. Samuelsson (1974) ArticleTitleProstaglandin endoperoxides. Novel transformations of arachidonic acid in human platelets Proc. Natl. Acad. Sci. USA 71 3400–3404 Occurrence Handle4215079 Occurrence Handle1:CAS:528:DyaE2MXitFCk

    PubMed  CAS  Google Scholar 

  • M. Hamberg C. Su E.H. Oliw (1998) ArticleTitleManganese lipoxygenase J. Biol. Chem. 21 13080–13088

    Google Scholar 

  • D.J. Hawkins T.G. Brash (1987) ArticleTitleEggs of the sea urchin, Strongylocentrotus purpuratuscontain a prominent (11R) and (12R) lipoxygenase activity J. Biol. Chem. 262 7629–7634 Occurrence Handle3108255 Occurrence Handle1:CAS:528:DyaL2sXksFKjtrg%3D

    PubMed  CAS  Google Scholar 

  • O. Heinisch E. Kowalski H. Ludwig N. Tauscher (1996) ArticleTitleStaining for soybean lipoxygenase activity in electrophoretic gels Fett/Lipid 98 183–184 Occurrence Handle1:CAS:528:DyaK28XktlWhtbY%3D

    CAS  Google Scholar 

  • H.L. Holland H.K. Weber (2003) ArticleTitleEnzymatic hydroxylation reaction Curr. Opin. Biotech. 11 547–553

    Google Scholar 

  • L. Hornsten S. Chao E. Osbourn U. Hellman E.H. Oliw (2002) ArticleTitleCloning of the manganese lipoxygenase gene reveals homology with the lipoxygenase gene family Eur. J. Biochem. 269 2690–2697 Occurrence Handle12047377 Occurrence Handle1:CAS:528:DC%2BD38XkslWkurc%3D

    PubMed  CAS  Google Scholar 

  • D. Iny A. Pinsky M. Cojocoru S. Grossman (1993a) ArticleTitleLipoxygenase of Thermoactinomyces vulgarispurification and characterization of reaction products Int. J. Biochem. 25 1313–1323 Occurrence Handle1:CAS:528:DyaK2cXis1WktA%3D%3D

    CAS  Google Scholar 

  • Huisman G.W. and Gray D. 2002. Towards novel processes for the fine-chemical industries. 13: 352-358.

  • D. Iny A. Pinsky S. Grossman (1993b) ArticleTitleLipoxygenase of Thermoactinomyces vulgaris-properties and study on the active site Int. J. Biochem. 25 1325–1330 Occurrence Handle1:CAS:528:DyaK3sXmt1Shs70%3D

    CAS  Google Scholar 

  • H. Kühn A. Borchert (2002) ArticleTitleRegulation of enzymatic peroxidation: the interplay of peroxidizing and peroxide reducing enzymes Free radical biol. med. 33 154–172

    Google Scholar 

  • H. Kuhn B.J. Thiele (1999) ArticleTitleThe diversity of the lipoxygenase family. Many sequences data but little information on biological significance FEBS Lett. 449 7–11 Occurrence Handle10225417 Occurrence Handle1:CAS:528:DyaK1MXis1arsbs%3D

    PubMed  CAS  Google Scholar 

  • C. Kyu C. Syn S. Swarup (2000) ArticleTitleA scalable protocol for the isolation of large-sized genomic DNA within and hour from a several bacteria Anal. Biochem. 278 86–90

    Google Scholar 

  • U.K. Laemmli (1970) ArticleTitleCleavage of structural proteins during the assembly of head of bacteriophageT4 Nature 277 680–685

    Google Scholar 

  • S.T. Prigge J.C. Boyington M. Faig K.S. Doctor B.J. Gaffney L.M. Amzel (1997) ArticleTitleStructure and mechanism of lipoxygenases Biochimie 79 629–636 Occurrence Handle9479444 Occurrence Handle1:CAS:528:DyaK1cXnvVehsw%3D%3D

    PubMed  CAS  Google Scholar 

  • J. Sambrook E.F. Fritsch T. Maniatis (1989) Molecular Cloning: A Laboratory Manual, 2nd ed Cold Spring Harbor Laboratory Press NY

    Google Scholar 

  • A. Schmid F. Hollmand J.B. Park B.b. Bülher (2002) ArticleTitleThe use of enzymes in the chemical industry in Europe Curr. Opin. Biotech. 13 359–366 Occurrence Handle12323359 Occurrence Handle1:CAS:528:DC%2BD38XmsFOnsLk%3D

    PubMed  CAS  Google Scholar 

  • D. Shibata H. Ohta (2002) Enzymatic transformation of lipids as a plant defense mechanism T.M. Kuo H.W. Gardner (Eds) Lipid Biotechnology Marcel Dekker New York 141–155

    Google Scholar 

  • Shimahara K. and Hashizume Y. 1973. Pseudomonas aeruginosa A-4 lipoxygenase-like enzyme. J. Ferm. Technol. 51: 183–189.

    Google Scholar 

  • C.K.P. Stover X.Q. Pham A.L. Erwin S.D. Mizoguci P. Warrener M.J. Hickey F.S. Brinkman W.O. Hufnagle D.J. Kowallk M. Lagrou R.L. Garber L. Goltry E. Tolentino S. Wesrbrock-Wadman Y. Yuan L.L. Brody S.N. Coulter K.R. Folger K.L. Kas R. Lim K. Smith D. Spencer G.K-S. Wong Z. Wu I.T. Paulsen J. Reizer M.H. Saler R.E.W. Hancock S. Lory M.W. Olson (2000) ArticleTitleComplete genome sequence of Pseudomonas aeruginosa PA01, an opportunistic pathogen Nature 406 959–964 Occurrence Handle10.1038/35023079 Occurrence Handle1:CAS:528:DC%2BD3cXmtlKqsrY%3D Occurrence Handle10984043

    Article  CAS  PubMed  Google Scholar 

  • C. Su E.H. Oliw (1998) ArticleTitleManganese lipoxygenase. Purification and characterization J. Biol. Chem 273 13072–13079 Occurrence Handle9582345 Occurrence Handle1:CAS:528:DyaK1cXjsVGis78%3D

    PubMed  CAS  Google Scholar 

  • P. Surrey (1964) ArticleTitleElectrophotometric method for determination of lipoxidase activity Plant Physiol. 39 65–70 Occurrence Handle10.1104/pp.39.1.65 Occurrence Handle16655881 Occurrence Handle1:CAS:528:DyaF2cXkt1Kmsb0%3D

    Article  PubMed  CAS  Google Scholar 

  • R.E. Vance S. Hong K. Gronet C. Serhan J.J. Mekalanos (2004) ArticleTitleThe opportunistic pathogen Pseudomnas aeruginosa carries a secretable arachidonate-15-lipoxygenase Proc. Natl. Acad. Sci. USA 101 2135–2139 Occurrence Handle14766977 Occurrence Handle1:CAS:528:DC%2BD2cXhs1Srs70%3D

    PubMed  CAS  Google Scholar 

  • D.C. Zimmerman B.A. Vick (1974) ArticleTitleLipoxygenase in Chlorella pyrenoidosa Lipids 8 264–266

    Google Scholar 

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Authors and Affiliations

  1. Laboratori de Microbiologia, Facultat de Farmàcia, Universitat de Barcelona, Barcelona, E-08028, Spain

    Jaume Vidal-Mas & Angeles Manresa

  2. Departament de Bioquimica i Biologia Molecular, Facultat de Quimica, Universitat de Barcelona, E-08028, Spain

    Montserrat Busquets

Authors
  1. Jaume Vidal-Mas
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  2. Montserrat Busquets
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  3. Angeles Manresa
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Correspondence to Angeles Manresa.

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Vidal-Mas, J., Busquets, M. & Manresa, A. Cloning and expression of a lipoxygenase from Pseudomonas aeruginosa 42A2. Antonie Van Leeuwenhoek 87, 245–251 (2005). https://doi.org/10.1007/s10482-004-4021-1

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  • DOI: https://doi.org/10.1007/s10482-004-4021-1

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