Heterologous Expression of Hybrid Type II Polyketide Synthase System in Streptomyces Species

  • Kim, Chang-Young (School of Chemical Engineering and Biotechnology, Inha University) ;
  • Park, Hyun-Joo (School of Chemical Engineering and Biotechnology, Inha University) ;
  • Kim, Eung-Soo (School of Chemical Engineering and Biotechnology, Inha University)
  • Published : 2003.10.01

Abstract

Polyketides are an extensive class of secondary metabolites with diverse molecular structures and biological activities. A plasmid-based minimal polyketide synthase (PKS) expression cassette was constructed using a subset of actinorhodin (act) biosynthetic genes (actI-orfl, actI-orf2, actI-orf3, actIII, actⅦ, and actIV) from Streptomyces coelicolor, which specify the construction of an orange-fluorescent anthraquinone product aloesaponarin II, a type II polyketide compound derived from one acetyl coenzyme A and 7 malonyl coenzyme A extender units. This system was designed as an indicator pathway in S. parvulus to generate a hybrid type II polyketide compound via gene-specific replacement. The act ${\beta}-ketoacyl$ synthase unit (actI-orfl and actI-orf2) in the expression cassette was specifically replaced with oxytetracycline ${\beta}-ketoacyl$ synthase otcY-orfl and otcY-orf2). This plasmid-based hybrid PKS cassette generated a novel orange-fluorescent compound structurally different from aloesaponarin II in both S. lividans and S. parvulus. In addition, several additional distinctive blue-fluorescent compounds were detected, when this hybrid PKS cassette was expressed in S. coelicolor B78 (actI-orf2 mutant), implying that the expression of plasmid-based hybrid PKS cassette in Streptomyces species should be an efficient way of generating hybrid type II polyketide compounds.

Keywords

References

  1. J. Bacteriol. v.172 Biosynthesis of anthraquinones by interspecies cloning of actinorhodin biosynthesis genes in streptomycetes: Clarification of actinorhodin gene functions Bartel,P.L.;C.B.Zhu;J.S.Lample;d.C.Dosch;N.C.Conners;W.R.Strohl;J.M.Beale;H.G.Floss
  2. Science v.156 Biosynthesis of polyketides and related compounds Birch,A.J. https://doi.org/10.1126/science.156.3772.202
  3. J. Microbiol. Biotechnol. v.12 Identification of Streptomyces sp. producing new polyene antibiotics and in vivo antimicrobial activity of tetrin C against phytopathogenic fungi Choi,W.C.;S.Y.Hwang;T.K.Park;S.K.Kim
  4. J. Chem. Soc. v.91 Derivatives of the multiple keten group Collie,J.N. https://doi.org/10.1039/ct9079101806
  5. Nature v.346 An unusually large multifunctional polypeptide in the erythromycin-producing polyketide synthase of Saccharopolyspora erythraea Cortes,J.;S.F.Haydock;G.A.Roberts;D.J.Bevitt;P.F.Leadlay
  6. J. Biol.Chem. v.267 Nucleotide sequences and deduced functions of a set of cotranslated genes of Streptomyces coelicolor A3(2) including the polyketide synthase for the antibiotic actinorhodin Fernandez-Moreno,M.A.;E.Martinez;L.Boto;D.A.Hopwood;F.Malpartida
  7. Nucleic Acids Res. v.16 A general method of in vitro preparation and specific mutagenesis of DNA fragments: Study of protein and DNA interactions Higuchi,R.B.;B.Krummel;R.K.Saiki https://doi.org/10.1093/nar/16.15.7351
  8. J. Microbiol. Biotechnol. v.11 Molecular cloning and characterization of the doxA cytochrome P-450 hydroxylase gene in Streptomyces peucetius subsp. caesius ATCC 27952 Hong,Y.S.;H.S.Kim;J.H.Lee;K.W.Kim;J.J.Lee
  9. Annu. Rev. Genet. v.24 Molecular genetics of polyketides and its comparison to fatty acid biosynthesis Hopwood,D.A.;D.H.Sherman https://doi.org/10.1146/annurev.ge.24.120190.000345
  10. Annu. Rev. Microbiol. v.49 Polyketide synthase gene manipulation: A structure-function approach in engineering novel antibiotics Hutchinson,C.R.;I.Fujii https://doi.org/10.1146/annurev.mi.49.100195.001221
  11. Annu. Rev. Microbiol. v.47 Polyketide synthesis: Prospects for hybrid antibiotics Katz,L.;S.Donadio https://doi.org/10.1146/annurev.mi.47.100193.004303
  12. Mol. Microbiol. v.6 Targeted gene replacements in a Streptomyces polyketide synthase gene cluster: Role for the acyl carrier protein Khosla,C.;S.Ebert-Khosla;D.A.Hopwood https://doi.org/10.1111/j.1365-2958.1992.tb01778.x
  13. J. Bacteriol. v.175 Genetic construction and functional analysis of hybrid polyketide synthases containing heterologous acyl carrier proteins Khosla,C.;R.McDaniel;S.Ebert-Khosla;R.Torres;D.H.Sherman;M.J.Bibb;D.A.Hopwood
  14. J. Microbiol. Biotechnol. v.11 Streptomyces showing antifungal activities against six plant pathogenic fungi Kim,B.J.;M.Cho;J.C.Kim;K.Y.Cho;G.J.Choi;C.H.Lee;Y.Lim
  15. J. Bacteriol. v.176 Analysis of type II polyketide β-ketoacyl synthase specificity in Streptomyces coelicolor A3(2) by trans complementation of actinorhodin synthase mutants Kim,E.S.;D.A.Hopwood;D.H.Sherman
  16. J. Bacteriol. v.177 Heterologous expression of an engineered biosynthetic pathway: Functional dissection of type II polyketide synthase components in Streptomyces species Kim,E.S.;K.D.Cramer;A.L.Shreve;D.H.Sherman
  17. Gene v.141 Sequences of the oxytetracycline polyketide synthase-encoding otc genes from Streptomyces rimosus Kim,E.S.;M.J.Bibb;M.J.Butler;D.A.Hopwood;D.H.Sherman https://doi.org/10.1016/0378-1119(94)90144-9
  18. Nature v.309 Molecular cloning of the whole biosynthetic pathway of a Streptomyces antibiotic and its expression in a heterologous host Malpartida,F.;D.A.Hopwood https://doi.org/10.1038/309462a0
  19. Mol. Gen. Genet. v.205 Physical and genetic characterization of the gene cluster for the antibiotic actinorhodin in Streptomyces coelicolor A3(2) Malpartida,F.;D.A.Hopwood https://doi.org/10.1007/BF02428033
  20. J. Am. Chem. Soc. v.115 Engineered biosynthesis of novel polyketides: Manipulation and analysis of an aromatic polyketide synthase with unproven catalytic specificities McDaniel,R.;S.Ebert-Khosla;D.A.Hopwood;C.Khosla https://doi.org/10.1021/ja00078a002
  21. Science v.262 Engineered biosynthesis of novel polyketides McDaniel,R.;S.Ebert-Khosla;D.A.Hopwood;C.Khosla https://doi.org/10.1126/science.8248802
  22. Nat. Prod. Rep. v.10 Biosynthesis of fatty acid and polyketide metabolites O'Hagan,D. https://doi.org/10.1039/np9931000593
  23. J. Bacteriol. v.174 Functional replacement of genes for individual polyketide synthase components in Streptomyces coelicolor A3(2) by heterologous genes from a different polyketide pathway Sherman,D.H.;E.S.Kim;M.J.Bibb;D.A.Hopwood