Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Molecular Docking and Kinetic Studies of the A226N Mutant of Deinococcus geothermalis Amylosucrase with Enhanced Transglucosylation Activity

  • Hong, Seungpyo (Research Group of Healthcare, Korea Food Research Institute) ;
  • Siziya, Inonge Noni (Department of Food Science and Technology, College of Agriculture and Life Sciences, Jeonbuk National University) ;
  • Seo, Myung-Ji (Division of Bioengineering, Incheon National University) ;
  • Park, Cheon-Seok (Graduate School of Biotechnology and Institute of Life Science and Resources, Kyung Hee University) ;
  • Seo, Dong-Ho (Department of Food Science and Technology, College of Agriculture and Life Sciences, Jeonbuk National University)
  • Received : 2020.03.30
  • Accepted : 2020.05.22
  • Published : 2020.09.28
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Abstract

Amylosucrase (ASase, E.C. 2.4.1.4) is capable of efficient glucose transfer from sucrose, acting as the sole donor molecule, to various functional acceptor compounds, such as polyphenols and flavonoids. An ASase variant from Deinococcus geothermalis, in which the 226th alanine is replaced with asparagine (DgAS-A226N), shows increased polymerization activity due to changes in the flexibility of the loop near the active site. In this study, we further investigated how the mutation modulates the enzymatic activity of DgAS using molecular dynamics and docking simulations to evaluate interactions between the enzyme and phenolic compounds. The computational analysis revealed that the A226N mutation could induce and stabilize structural changes near the substrate-binding site to increase glucose transfer efficiency to phenolic compounds. Kinetic parameters of DgAS-A226N and WT DgAS were determined with sucrose and 4-methylumbelliferone (MU) as donor and acceptor molecules, respectively. The kcat/Km value of DgAS-A226N with MU (6.352 mM-1min-1) was significantly higher than that of DgAS (5.296 mM-1min-1). The enzymatic activity was tested with a small phenolic compound, hydroquinone, and there was a 1.4-fold increase in α-arbutin production. From the results of the study, it was concluded that DgAS-A226N has improved acceptor specificity toward small phenolic compounds by way of stabilizing the active conformation of these compounds.

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References

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