Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Investigations on Possible Roles of C-Terminal Propeptide of a Ca-Independent ${\alpha}$-Amylase from Bacillus

  • Salimi, Ali (Department of Biochemistry, Faculty of Biological Science, Tarbiat Modares University) ;
  • Yousefi, Fatemeh (Department of Biochemistry, Faculty of Biological Science, Tarbiat Modares University) ;
  • Ghollasi, Marzieh (Department of Biochemistry, Faculty of Biological Science, Tarbiat Modares University) ;
  • Daneshjou, Sara (Department of Biochemistry, Faculty of Biological Science, Tarbiat Modares University) ;
  • Tavoli, Hesam (Department of Biochemistry, Faculty of Biological Science, Tarbiat Modares University) ;
  • Ghobadi, Sirous (Department of Biology, Faculty of Science, Razi University of Kermanshah) ;
  • Khajeh, Khosro (Department of Biochemistry, Faculty of Biological Science, Tarbiat Modares University)
  • Received : 2011.12.07
  • Accepted : 2012.04.06
  • Published : 2012.08.28
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Abstract

Previously, an extracellular ${\alpha}$-amylase (BKA) had been purified from the culture of Bacillus sp. KR8104. Subsequently, the crystal structure of the active enzyme revealed a 422 amino acids polypeptide. In this study, the bka was cloned into E. coli, which encoded a polypeptide of 659 amino acids including two additional fragments: one 44 residues N-terminal fragment and another 193 residues C-terminal fragment. In order to investigate the role of the C-terminal fragment, two constructs with and without this region [$BKA{\Delta}$(N44) and $BKA{\Delta}$(N44C193)] were designed and expressed in E. coli BL21. The optimum pH, thermal stability, and the end-products of starch hydrolysis were found to be similar in both constructs. The $K_m$ and $V_{max}$ values for $BKA{\Delta}$(N44) were lower than $BKA{\Delta}$(N44C193), using either starch or ethylidene-blocked 4-nitrophenylmaltoheptaoside as a substrate.

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References

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