Journal of Microbiology and Biotechnology

  • 제26권10호
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  • Pages.1717-1722
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  • 2016
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  • 1017-7825(pISSN)
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  • 1738-8872(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
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Enhancing the Thermal Resistance of a Novel Acidobacteria-Derived Phytase by Engineering of Disulfide BridgesS

  • Tan, Hao (Soil and Fertilizer Institute, Sichuan Academy of Agricultural Sciences) ;
  • Miao, Renyun (Soil and Fertilizer Institute, Sichuan Academy of Agricultural Sciences) ;
  • Liu, Tianhai (Soil and Fertilizer Institute, Sichuan Academy of Agricultural Sciences) ;
  • Cao, Xuelian (Soil and Fertilizer Institute, Sichuan Academy of Agricultural Sciences) ;
  • Wu, Xiang (Soil and Fertilizer Institute, Sichuan Academy of Agricultural Sciences) ;
  • Xie, Liyuan (Soil and Fertilizer Institute, Sichuan Academy of Agricultural Sciences) ;
  • Huang, Zhongqian (Soil and Fertilizer Institute, Sichuan Academy of Agricultural Sciences) ;
  • Peng, Weihong (Soil and Fertilizer Institute, Sichuan Academy of Agricultural Sciences) ;
  • Gan, Bingcheng (Soil and Fertilizer Institute, Sichuan Academy of Agricultural Sciences)
  • 투고 : 2016.04.19
  • 심사 : 2016.06.29
  • 발행 : 2016.10.28
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초록

A novel phytase of Acidobacteria was identified from a soil metagenome, cloned, overexpressed, and purified. It has low sequence similarity (<44%) to all the known phytases. At the optimum pH (2.5), the phytase shows an activity level of 1,792 μmol/min/mg at physiological temperature (37℃) and could retain 92% residual activity after 30 min, indicating the phytase is acidophilic and acidostable. However the phytase shows poor stability at high temperatures. To improve its thermal resistance, the enzyme was redesigned using Disulfide by Design 2.0, introducing four additional disulfide bridges. The half-life time of the engineered phytase at 60℃ and 80℃, respectively, is 3.0× and 2.8× longer than the wild-type, and its activity and acidostability are not significantly affected.

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