2016 ; Vol.26-10: 1717~1722
|Author||Hao Tan, Renyun Miao, Tianhai Liu, Xuelian Cao, Xiang Wu, Liyuan Xie, Zhongqian Huang, Weihong Peng, Bingcheng Gan|
|Place of duty||Soil and Fertilizer Institute, Sichuan Academy of Agricultural Sciences, Chengdu 610066, P.R. China,Scientific Observing and Experimental Station of Agro-microbial Resource and Utilization in Southwest China, Ministry of Agriculture, Chengdu 610066, P.R. China|
|Title||Enhancing the Thermal Resistance of a Novel Acidobacteria-Derived Phytase by Engineering of Disulfide Bridges|
J. Microbiol. Biotechnol.2016 ;
|Abstract||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.|
|Key_word||phytase, acidophilic, acidostable, site-directed mutagenesis, disulfide bridge, thermostable|
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