Cloning of Biosurfactant-Producing Gene from Bacillus subtilis KL-57

Bacillus subtilis KL-57로부터 생산되는 생체계면활성제 합성 유전자 클로닝

  • 강상모 (건국대학교 공과대학 미생물공학과) ;
  • 이병옥 (건국대학교 공과대학 미생물공학과) ;
  • 이철수 (건국대학교 공과대학 미생물공학과)
  • Published : 1994.12.01


A bacterium KL-57 which exhibited biosurfactant activity was isolated. This bacterium was identified as Bacillus subtilis. The biosurfactant-producing gene of B. subtilis KL-57 was cloned into R subtilis MI113 by using plasmid pTB523. The plasmid DNA from the clone was found to carry a 18 kb PstI insert. The biosurfactant-producing gene was cleaved into 4 fragments by SmaI, 3 fragments by PvulI or EcoRl, 4 fragments by PvulI and EcoRI double digestion, 5 fragments by AccI, and 2 fragments by KpnI, HindIII or BamHI. By subcloning the 18 kb Pstl insert, a 2.3 kb EcoRl fragment conferred the biosurfactant producing activity on B. subtilis cells. The 2.3 kb had one HindIII cleave site. But Two fragments, which corresponds HindIII/EcoRl termini, exhibited no biosurfactant activity.


Biosurfactant;Bacilluus subtilis;surfactin;gene cloning


  1. Biosurfactant and Biotechnology Kosaric,N.;W.L.Carins
  2. J. Bacteriol v.171 Gene cluster containing the genes for tyrocidine synthetase 1 and 2 from Bacillus brevis : evidence for an operon Mittenhuber,G.;R.Weckermann;M.A.Marahiel
  3. Chem. Pharm. Bull. v.22 Antitumor activity of Bacillus natto. V. Isolation and characterization of surfactin in the culture medium of Bacillus natto KMD23111 Kameda,Y.;S.Ohira;K.Matsui;S.Kanatomo;T.Hase;T.Atsusaka
  4. J. Bacteriol. v.171 Cloning and characterization of srfB, a regulatory gene involved in surfactin production and competence in Bacillus subtilis Nakano,M.M.;P.Zuber
  5. J. Bacteriol v.173 srfA is an operon required for surfactin production, competence developement, and efficient sporulation in Bacillus subtilis Nakano,M.M.;R.Magnuson;A.Myers;P.Zuber
  6. 산업미생물학회지 v.22 Biosurfactant를 생산하는 P. aeruginosa. KK-7의 분리 및 biosurfactant의 생산 강상모;김대원;김혜자
  7. J. Ferment. Bioeng. v.75 Survival of Bacillus subtilis NB22, an antifungal-antibiotic iturin producer, and its transformation in soil-system Yukinori,T.;A.Takashi;S.Makoto
  8. Adv. Appl. Microbiol. v.26 Surface-active compound from microorganisms Cooper,D.G.;J.E.Zarjic
  9. J. Ferment. Bioeng. v.69 Suppressive effect of Bacillus subtilis and it's products on phytopathogenic microorganisms Phae,C.G.;M.Shoda;H.Kubota
  10. J. Bacteriol. v.146 Isolation and characterization of antibiotic resistance plasmids from thermophilic bacilli and construction of deletion plasmids Imanaka,T.;M.Fujii;S.Aiba
  11. Biotechnol. Lett. v.12 Transformation of Bacillus subtilis with the treatment by alkali cations Ano,T.;A.Kobayashi;M.Shoda
  12. J. Ferment. Bioeng. v.74 Molecular cloning of a gene responsible for the biosynthesis of the lipopeptide antibiotics iturin and surfactin Hiraoka,H.;T.Ano;M.Shoda
  13. Molecular cloning, a laboratory manual Maniatis,T.;E.F.Fritsch;J.Sambrook
  14. J. Ferment. Bioeng. v.74 Rapid transformation of Bacillus subtilis using KCI-treatment Hiraoka,H.;T.Ano;M.Shoda
  15. J. Bacteriol. v.81 Requirement for transformation in Bacillus subtilis, and Bacillus licheniformis Anagnostopoulos,C.;J.Spizizen
  16. In Bergey's Manual of Systematic Bacteriology v.2 Genus Bacillus Cohn Sneath,P.A.;N.S.Mair;M.E.Sharpe;J.G.Holt
  17. 한국공업규격 합성세제 시험범 KSM 2702, M2709-1991 한국공업표준협회
  18. J. Bacteriol. v.169 Cloning and nucleotide sequence of the penicillinase anti-repressor gene of Bacillus licheniformis Imanaka,T.;T.Himeno;S.Aiba
  19. J. Ferment. Bioeng. v.74 Isolation of a new surfactin producer Bacillus pumilus A-1, and cloning and Nucleotide sequence of the regulator gene, psf-1 Masaaki,M.;I.Mikito;I.Tadayuki
  20. Biochem. Biophys. Res. Commun. v.31 Surfactin, a crystalline peptidelipid surfactant produced by Bacillus subtilis : isolation, characterization and its inhibition of fibrin clot formation Arima,K.;A.Kakimura;G.Tamura
  21. J. Bacteriol. v.170 Identification of a genetic locus required for biosynthesis of the lipopeptide antibiotic surfactin in Bacillus subtilis Nakano,M.M.;M.A.Marahiel;P.Zuber
  22. Molec. Gen. Genet. v.168 High frequency transformation of Bacillus subtilis protoplasts by plasmid DNA Chang,S.;S.N.Cohen
  23. Molec. Gen. Genet. v.232 Isolation and characterization of sfp;a gene that functions in the production of the lipopeptide biosurfactant, surfactin, in Bacillus subtilis Nakano,M.;N.Corbell;J.Besson;P.Zuber
  24. Molecular Cloning: A Laboratory Manual(2nd ed.) Sambrook,J.;E.F.Fritsch;T.Maniatis
  25. Anal. Biochem. v.114 A rapid method for the precipitation of bacterial plasmids Holmes,D.S.;M.Quigley
  26. J. Bacteriol. v.171 Gramicidin S biosynthesis operon containing the structural genes grsA and grsB has an openreading frame encoding a protein homologous to fatty acid thioesterases Kratzschmar,J.;M.Krause;M.A.Marahiel
  27. Molecular Cloning : A Labortory Maual(2nd ed.) Sambrook,J.;E.F.Fritsch;T.Maniatis
  28. J. Bacteriol. v.169 Cloning and nucleotide sequence of the penicillinase antirepressor gene of Bacillus licheniformis Imanaka,T.;M.Kuroda;K.Sakaguchi
  29. Nucleic Acids Res. v.7 A rapid alkaline extraction procedure for screening recombinant plasmid DNA Birhboim,H.C.;J.Doly
  30. Method enzymol v.101 New M13 vectors for cloning Messing,J.
  31. 黴生物の分離と同定, 好氣性細菌 長谷川武治