Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Effect of Ion Pair on Thermostability of F1 Protease: Integration of Computational and Experimental Approaches

  • Rahman, Raja Noor Zaliha Raja Abd (Enzyme and Microbial Technology Research Group, Faculty of Biotechnology and Biomolecular Sciences, University Putra Malaysia) ;
  • Noor, Noor Dina Muhd (Enzyme and Microbial Technology Research Group, Faculty of Biotechnology and Biomolecular Sciences, University Putra Malaysia) ;
  • Ibrahim, Noor Azlina (Enzyme and Microbial Technology Research Group, Faculty of Biotechnology and Biomolecular Sciences, University Putra Malaysia) ;
  • Salleh, Abu Bakar (Enzyme and Microbial Technology Research Group, Faculty of Biotechnology and Biomolecular Sciences, University Putra Malaysia) ;
  • Basri, Mahiran (Enzyme and Microbial Technology Research Group, Faculty of Science, University Putra Malaysia)
  • Received : 2011.05.27
  • Accepted : 2011.09.15
  • Published : 2012.01.28
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Abstract

A thermophilic Bacillus stearothermophilus F1 produces an extremely thermostable serine protease. The F1 protease sequence was used to predict its three-dimensional (3D) structure to provide better insights into the relationship between the protein structure and biological function and to identify opportunities for protein engineering. The final model was evaluated to ensure its accuracy using three independent methods: Procheck, Verify3D, and Errat. The predicted 3D structure of F1 protease was compared with the crystal structure of serine proteases from mesophilic bacteria and archaea, and led to the identification of features that were related to protein stabilization. Higher thermostability correlated with an increased number of residues that were involved in ion pairs or networks of ion pairs. Therefore, the mutants W200R and D58S were designed using site-directed mutagenesis to investigate F1 protease stability. The effects of addition and disruption of ion pair networks on the activity and various stabilities of mutant F1 proteases were compared with those of the wild-type F1 protease.

Keywords

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