Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Characterization of a Bifunctional HPr Kinase/Phosphorylase from Leuconostoc mesenteroides SY1

  • Park, Jae-Yong (Institute of Agriculture & Life Science, Gyeongsang National University) ;
  • Lee, Kang-Wook (Division of Applied Life Science (BK21), Graduate School, Gyeongsang National University) ;
  • Lee, Ae-Ran (Division of Applied Life Science (BK21), Graduate School, Gyeongsang National University) ;
  • Jeong, Woo-Ju (Division of Applied Life Science (BK21), Graduate School, Gyeongsang National University) ;
  • Chun, Ji-Yeon (Department of Food Science and Technology, Sunchon National University) ;
  • Lee, Jong-Hoon (Department of Foods and Biotechnology, Kyunggi University) ;
  • Kim, Jeong-Hwan (Division of Applied Life Science (BK21), Graduate School, Gyeongsang National University)
  • Published : 2008.04.30
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Abstract

The hprK gene encoding bifunctional HPrK/P (kinase/phosphorylase) was cloned from L. mesenteroides SY1, a strain isolated from kimchi. hprK was transcribed as a monocistronic gene. His-tagged HPrH16A and HPrK/P were produced in E. coli BL21 (DE3) using pET26b(+) and purified. HPrK/P phosphorylation assay with purified proteins showed that the kinase activity of HPrK/P increased at slightly acidic pHs. Divalent cations such as $Mg^{2+}$ and $Mn^{2+}$ and glycolytic intermediates such as fructose-1, 6-bisphosphate (FBP) and phosphoenolpyruvate (PEP) increased the kinase activity of HPrK/P, but inorganic phosphate strongly inhibited it. Kinetic studies for the kinase activity of HPrK/P showed that the apparent $K_m$ values were 0.18 and $14.57{\mu}M$ for ATP and HPr, respectively. The $K_m$ value for the phosphorylase activity of HPrK/P was $14.16{\mu}M$ for P-Ser-HPr (HPr phosphorylated at the serine residue).

Keywords

References

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