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Characterization of the Starch Degradation Activity of recombinant glucoamylase from Extremophile Deinococcus geothermalis

극한성 미생물Deinococcus geothermalis 유래 재조합 글루코아밀레이즈의 전분 분해 활성 특징

  • Jang, Seung-Won (ForBioKorea Co., Ltd.) ;
  • Kwon, Deok-Ho (Department of Bioengineering and Technology, Kangwon National University) ;
  • Park, Jae-Bum (Department of Bioengineering and Technology, Kangwon National University) ;
  • Jung, Jong-Hyun (Research Division for Biotechnology, Korea Atomic Energy Research Institute) ;
  • Ha, Suk-Jin (Department of Bioengineering and Technology, Kangwon National University)
  • Received : 2019.09.17
  • Accepted : 2019.10.07
  • Published : 2019.10.31

Abstract

This work focused on characterization of the starch degradation activity from extremophile strain Deinococcus geothermalis. Glucoamylase gene from D. geothermalis was cloned and overexpressed by pET-21a vector using E. coli BL21 (DE3). In order to characterize starch degrading activity of recombinant glucoamylase, enzyme was purified using HisPur Ni-NTA column. The recombinant glucoamylase from D. geothermalis exhibited the optimum temperature as $45^{\circ}C$ for starch degradation activity. And highly acido-stable starch degrading activity was shown at pH 2. For further optimization of starch degrading activity with metal ion, various metal ions ($AgCl_2$, $HgCl_2$, $MnSO_4{\cdot}4H_2O$, $CoCl_2{\cdot}6H_2O$, $MgSO_4$, $ZnSO_4{\cdot}7H_2O$, $K_2SO_4$, $FeCl_2{\cdot}4H_2O$, NaCl, or $CuSO_4$) were added for enzyme reaction. As results, it was found that $FeCl_2{\cdot}4H_2O$ or $MnSO_4{\cdot}4H_2O$ addition resulted in 17% and 9% improved starch degrading activity, respectively. The recombinant glucoamylase from D. geothermalis might be used for simultaneous saccharification and fermentation (SSF) process at high acidic conditions.

Keywords

References

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