Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지
DOI QR코드

DOI QR Code

Characterization of Dephosphocoenzyme A Kinase from Streptomyces peucetius ATCC27952, and Its Application for Doxorubicin Overproduction

  • Lee, Na-Rae (Department of Pharmaceutical Engineering, SunMoon University) ;
  • Rimal, Hemraj (Department of Pharmaceutical Engineering, SunMoon University) ;
  • Lee, Joo-Ho (Department of Pharmaceutical Engineering, SunMoon University) ;
  • Oh, Tae-Jin (Department of Pharmaceutical Engineering, SunMoon University)
  • Received : 2014.04.29
  • Accepted : 2014.07.15
  • Published : 2014.09.28
Download PDF
⟨ Previous Next ⟩

Abstract

Dephosphocoenzyme A (CoaE) catalyzes the last step in the biosynthesis of the cofactor coenzyme A. In this study, we report the identification and application of CoaE from Stretomyces peucetius ATCC27952. After expression of coaE, the protein was found to have a molecular mass of 28.6 kDa. Purification of the His-tagged fused CoaE protein was done by immobilized metal-affinity chromatography, and then in vitro enzymatic coupling assay was performed. The increasing NADH consumption with time shed light on the phosphorylating activity of CoaE. Furthermore, the overexpression of coaA and coaE independently under the $ermE^*$ promoter in the doxorubicin -producing wild type strain, resulted in 1.4- and 1.5-fold enhancements in doxorubicin production, respectively. In addition, the overexpression of both genes together showed a 2.1-fold increase in doxorubicin production. These results established a positive role for secondary metabolite production from Streptomyces peucetius.

Keywords

References

  1. Altschul SF, Gish W, Miller W, Myss EW, Lipman DJ. 1990. Basic local alignment search tool. J. Mol. Biol. 215: 403-410. https://doi.org/10.1016/S0022-2836(05)80360-2
  2. Begley TP, Kinslnad C, Taylor S, Tandon M, Nicewonger R, Wu M, et al. 1998. Cofactor biosynthesis: a mechanistic perspective. Top. Curr. Chem. 195: 93-142. https://doi.org/10.1007/3-540-69542-7_3
  3. Bradford MM. 1976. A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248-254. https://doi.org/10.1016/0003-2697(76)90527-3
  4. Daugherty M, Polanuyer B, Farrell M, Scholle M, Lykidis A, de Crecy-Lagard V, Osterman A. 2002. Complete reconstitution of the human coenzyme A biosynthetic pathway via comparative genomics. J. Biol. Chem. 277: 21431-21439. https://doi.org/10.1074/jbc.M201708200
  5. Geerlof A, Lewendon A, Shaw WV. 1999. Purification and characterization of phosphopantetheine adenyltransferase from Escherichia coli. J. Biol. Chem. 274: 27105-27111. https://doi.org/10.1074/jbc.274.38.27105
  6. Halvorsen O, Skrede S. 1982. Regulation of the biosynthesis of CoA at the level of pantothenate kinase. Eur. J. Biochem. 124: 211-215. https://doi.org/10.1111/j.1432-1033.1982.tb05927.x
  7. Kieser T, Bibb MJ, Buttner MJ, Chater KF, Hopwood DA. 2000. Practical Streptomyces Genetics. The John Innes Centre Foundation, Norwich.
  8. Kleinkauf H. 2000. The role of 4'-phosphopantethine in the biosynthesis of fatty acids, polyketides and peptides. Biofactors 11: 91-92. https://doi.org/10.1002/biof.5520110126
  9. Kupke T. 2002. Molecular characterization of the 4- phosphopantothenoylcysteine synthase domain of bacterial Dfp flavoproteins. J. Biol. Chem. 277: 36137-36145. https://doi.org/10.1074/jbc.M206188200
  10. Leonardi R, Zhang Y, Charles OR, Jackowski S. 2005. Coenzyme A: back in action. Prog. Lipid Res. 44: 125-153. https://doi.org/10.1016/j.plipres.2005.04.001
  11. Mandakh A, Niraula NP, Kim EP, Sohng JK. 2010. Identification and characterization of a pantothenate kinase (PanK-sp) from Streptomyces peucetius ATCC 27952. J. Microbiol. Biotechnol. 20: 1689-1695. https://doi.org/10.4014/jmb.1007.07058
  12. Mishra PK, Park PK, Drueckhammer DG. 2001. Identification of yacE (coaE) as the structural gene for dephosphocoenzyme A kinase in Escherichia coli K-12. J. Bacteriol. 183: 2774-2778. https://doi.org/10.1128/JB.183.9.2774-2778.2001
  13. Olzhausen J, Schubbe S, Schuller HJ. 2009. Genetic analysis of coenzyme A biosynthesis in the yeast Saccharomyces cerevisiae: identification of a conditional mutation in the pantothenate kinase gene CAB1. Curr. Genet. 55: 163-173. https://doi.org/10.1007/s00294-009-0234-1
  14. Pfeifer BA, Admiraal SJ, Gramajo H, Cane DE, Khosla C. 2001. Biosynthesis of complex polyketides in a metabolically engineered strain of E. coli. 291: 1790-1792. https://doi.org/10.1126/science.1058092
  15. Robinshaw JD, Neely JR. 1985. Coenzyme A metabolism. Am. J. Physiol. Endocrinol. Metab. 248: E1-E9. https://doi.org/10.1152/ajpendo.1985.248.1.E1
  16. Sambrook J, Russell DW. 2001. Molecular Cloning: A Laboratory Manual. 3rd Ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
  17. Sambrook J, Fritsch EF, Maniatis T. 1989. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
  18. Song WJ, Jackowcki S. 1992. Cloning, sequencing, and expression of the pantothenate kinase (coaA) gene of Escherichia coli. J. Bacteriol. 174: 6411-6417. https://doi.org/10.1128/jb.174.20.6411-6417.1992
  19. Spry C, Kirk K, Saliba KJ. 2008. Coenzyme A biosynthesis: an antimicrobial drug target. FEMS Microbiol Rev. 32: 56-106. https://doi.org/10.1111/j.1574-6976.2007.00093.x
  20. Sthapit B, Oh T-J, Lamichhance R, Liou K, Lee HC, Kim CG, Sohng JK. 2004. Neocarzinostatin naphthoate synthase: a unique iterative type I PKS from neocarzinostatin producer Streptomyces carzinostaticus. FEBS Lett. 656: 201-206.
  21. Vadali RV, Bennett GN, San K. 2004. Cofactor engineering of intracelluar CoA/acetyl-CoA and its effect on metabolic flux redistribution in Escherichia coli. Metab. Eng. 6: 133-139. https://doi.org/10.1016/j.ymben.2004.02.001
  22. Vallaari DS, Jackowski S, Rock CO. 1987. Regulation of pantothenate kinase by coenzyme A and its thioesters. J. Biol. Chem. 262: 2468-2471.
  23. Walia G, Surolia A. 2011. Insights into the regulatory characteristics of the mycobacterial dephosphocoenzyme A kinase: implications for the universal CoA biosynthesis pathway. PLoS One 6: e21390. https://doi.org/10.1371/journal.pone.0021390

Cited by

  1. Genome-guided exploration of metabolic features of Streptomyces peucetius ATCC 27952: past, current, and prospect vol.102, pp.10, 2018, https://doi.org/10.1007/s00253-018-8957-x