Construction of the Recombinant phbCAB Operon of Alcaligenes eutvtrphus for Accumulation of Poly-$\beta$-hydroxybu tyric Acid in Escherichia coli

Alcaligenes eutrophus phbCAB Operon의 재조합과 Poly-$\beta$-hydroxybutyric Aicd의 대장균내 축적

  • 김경태 (경북대학교 자연과학대학 유전공학과) ;
  • 박진서 (경북대학교 자연과학대학 유전공학과) ;
  • 이용현 (경북대학교 자연과학대학 유전공학과) ;
  • 허태린 (경북대학교 자연과학대학 유전공학과) ;
  • 박해철 (경북대학교 자연과학대학 유전공학과)
  • Published : 1993.06.01


In order to achieve poly-beta-hydroxybutyric acid (PHB) production using recombinant DNA in various host bacterial cells, the isolation of genes for PHB biosynthesis was attempted. As a result, a 5.2kb DNA fragment containing phbCAB operon of Alcaligenes eutrophus was isolated by colony hybridization using synthetic oligodeoxyribonucleotides as probes. The constructed recmbinant plasmid pSK(+)-phbCAB operon was transferred to Escherichia coli, and the obtained transformant accumulated considerable amount of PHB.


  1. Appl. Environ. Microbiol. v.32 Quantitative requirements for exponential growth of alcaligenes eutrophus. Repaske,R.;A.C.Repaske
  2. J. Biol. Chem. v.264 Poly-β-hydroxybutyrate(PHB)biosynthesis and characterization of the PHB polymerase gene(phbC) Peoples.O.P.;A.J.Sinskey
  3. Appl. Microbiol. Biotechnol. v.27 Control of molecular weight of poly-β-hydroxybutytic acid produced in fedbatch cuiture of Protomonas extorquens Suzuki,T.;H.Deguchi.;T.Yamane;S.Shimizu;K.Gekko
  4. Biotechnol. Lett. v.11 A survey of the accumulation of novel polyhydroxyalkanoate by Bacteria. Haywood,G.W.;A.J.Anderson;E.H.Dawes
  5. Meth. Enzymol. v.68 Plasmid of E. coil as cloning vector Bolivar,F.;K.Backman
  6. Nucleric Acids Res. v.16 λZAP:A bacteriophage λ expression vector with in vivo execision properties Short,J.M.;J.M.Fernandez;J.A.Sorge;W.D.Huse.
  7. Molecular Cloning. A Laboratorry Manual. (2nd ed.) Sambrook,J.;E.Fritsch;T.Manatis
  8. Appl. Microbiol. Biotchnol. v.28 Production of copolyesters of`3-hydroxybutyrate and 3-hydroxyvalerate by Alcaligenes eutrophus from butyric and pentanoic acids Doi,Y.;A.Tamaki;M.Kunioka;K.Soga
  9. Gene Cloning. Brown,T.A.
  10. Nucleic Acids Res. v.7 A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Birthoim,H.C.;J.Doly.
  11. Molecular Cloning. A Laboratorry Manual. (1sr ed.) Sambrook,J.;E.Fritsch;T.Manatis
  12. Proc. Natl. Acad. Sci. USA v.74 DNA sequencing with chain terminating ingibitors Sanger,G.;S.Nicklen;A.R.Coulson
  13. J. Biol. Chem. v.264 Poly-β-hydroxybutyrate biosynthesis in Alcaligenes cutrophus H16:characterization of the genes encoding β-ketothiolase and acetoacetyl-CoA reductase Peoples.O.P.;A.J.Sinskey
  14. The enzymes v.14 Bacteriophage T4 polynicleotide kinase Richardson.C.C.;P.D.Boyer(Ed.)
  15. J. Mol. Biol. v.88 Detection of specific sequences among DNA fragments separated by gel electrophoresis Southern,E.
  16. Proc. Natl. Acad. Sci. USA v.72 Colony hybridization:a method for the isolation of clonned DNAs that contined a specific gene Grunstein,M.;D.S.Hogness
  17. J. Bacteriol. v.82 Assay of Poly β-hydroxybutyric acid. Law,J.H.;R.A.Slepecky
  18. Anal. Chem. v.31 Use of dinitrosalicylic acid reagent for detemination of reducing sugar Miller,G.L.
  19. J. Bacteriol. v.170 Cloning and expression in Escherichia coli of the Alcaligenes eurtrophus H16 poly-β-hydroxybutyric biosynthetic pathway. Slater,S.C.;W.H.Voige;D.E.Dennis