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( Hao Tan ),( Renyun Miao ),( Tianhai Liu ),( Xuelian Cao ),( Xiang Wu ),( Liyuan Xie ),( Zhongqian Huang ),( Weihong Peng ),( Bingcheng Gan ) 한국미생물 · 생명공학회 2016 Journal of microbiology and biotechnology Vol.26 No.10
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°C) 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°C and 80°C, respectively, is 3.0× and 2.8× longer than the wild-type, and its activity and acidostability are not significantly affected.