title: Enhancing the Thermal Resistance of a Novel Acidobacteria-Derived Phytase by Engineering of Disulfide Bridges.
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.
authors.affiliation: 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.
authors.forename: Hao
authors.initials: H
authors.lastname: Tan
authors.affiliation: 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.
authors.forename: Renyun
authors.initials: R
authors.lastname: Miao
authors.affiliation: 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.
authors.forename: Tianhai
authors.initials: T
authors.lastname: Liu
authors.affiliation: 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.
authors.forename: Xuelian
authors.initials: X
authors.lastname: Cao
authors.affiliation: 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.
authors.forename: Xiang
authors.initials: X
authors.lastname: Wu
authors.affiliation: 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.
authors.forename: Liyuan
authors.initials: L
authors.lastname: Xie
authors.affiliation: 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.
authors.forename: Zhongqian
authors.initials: Z
authors.lastname: Huang
authors.affiliation: 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.
authors.forename: Weihong
authors.initials: W
authors.lastname: Peng
authors.affiliation: 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.
authors.forename: Bingcheng
authors.initials: B
authors.lastname: Gan
mesh_terms: D010832:6-Phytase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D061271:Acidobacteria / Q000201:enzymology / Q000235:genetics*; D004220:Disulfides / Q000737:chemistry* / Q000378:metabolism; D004926:Escherichia coli; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D004220:Disulfides; D011994:Recombinant Proteins; D010832:6-Phytase
keywords: Phytase; acidophilic; acidostable; disulfide bridge; site-directed mutagenesis; thermostable