Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Identification and Characterization of a Pantothenate Kinase (PanK-sp) from Streptomyces peucetius ATCC 27952

  • Mandakh, Ariungerel (Institute of Biomolecule Reconstruction (iBR), Department of Pharmaceutical Engineering, Sun Moon University) ;
  • Niraula, Narayan Prasad (Institute of Biomolecule Reconstruction (iBR), Department of Pharmaceutical Engineering, Sun Moon University) ;
  • Kim, Eung-Pil (Institute of Biomolecule Reconstruction (iBR), Department of Pharmaceutical Engineering, Sun Moon University) ;
  • Sohng, Jae-Kyung (Institute of Biomolecule Reconstruction (iBR), Department of Pharmaceutical Engineering, Sun Moon University)
  • Received : 2010.07.30
  • Accepted : 2010.09.01
  • Published : 2010.12.28
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Abstract

Pantothenate kinase (PanK) catalyzes the first step in the biosynthesis of the essential and ubiquitous cofactor coenzyme A (CoA) in all organisms. Here, we report the identification, cloning, and characterization of panK-sp from Streptomyces peucetius ATCC 27952. The gene encoded a protein of 332 amino acids with a calculated molecular mass of 36.8 kDa and high homology with PanK from S. avermitilis and S. coelicolor A3(2). To elucidate the putative function of PanK-sp, it was cloned into pET32a(+) to construct pPKSP32, and the PanK-sp was then expressed in E. coli BL21(DE3) as a His-tag fusion protein and purified by immobilized metal affinity chromatography. The enzyme assay of PanK-sp was carried out as a coupling assay. The gradual decrease in NADH concentration with time clearly indicated the phosphorylating activity of PanK-sp. Furthermore, the ca. 1.4-fold increase of DXR and the ca. 1.5-fold increase of actinorhodin by in vivo overexpression of panK-sp, constructed in pIBR25 under the control of a strong $ermE^*$ promoter, established its positive role in secondary metabolite production from S. peucetius and S. coelicolor, respectively.

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References

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