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《Journal of Hubei University(Natural Science)》 2018-01
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Display expression of a lipase from Proteus sp. on the Escherichia coli cell surface and characterization of the recombinant enzyme

TIAN Shufang;WANG Yu;LI Chunhua;Hubei Key Laboratory of Industrial Biotechnology,Hubei Collaborative Innovation Center for Green Transformation of Bio-resources,College of Life Sciences,Hubei University;  
Background for industrial bioconversion processes,the utilization of surface-displayed lipase in the form of whole-cell biocatalysts is more advantageous,because the enzymes are displayed on the cell surface spontaneously,regarded as immobilized enzymes.The carboxyl terminal domain of sf GFP(super fold green fluorescent protein)was used as carrier protein and the Lip A gene from Proteus sp.was fused to the sf GFP gene.The fusion gene sf GFP-Lip A was inserted the plasmid p ETsf GFP,which was named as p ETsf GFP-Lip A.The recombinant plasmid was transformed into E.coli Rosetta-Blue host after self-induced expression.Localization of the displayed Lip A on the cell surface was confirmed by the confocal laser scanning microscopy and flow cytometry.The activity of the recombinant Lip A has reached to 7 009 U/g dry cells,and the optimal temperature and p H of lipase was 37℃and 8.0,respectively.The activity was significantly increased in the presence of the metal ions such as Co~(2+)and Ni~(2+)and Tirton X-100.It was indicated that the recombinant Lip A exhibited resistance to organic solvents after the treatment with 60%acetone and 60%ethanol for 18 h,the activity was increased by 104%and 117%,respectively.So the above results indicate that the advantage of surface-displayed lipase could lay a solid foundation for industrial application.
【Fund】: 国家自然科学基金(31100124);; 湖北省教育厅重点项目(D20131004)资助
【CateGory Index】: Q55
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