JOURNAL OF SHANDONG UNIVERSITY(NATURAL SCIENCE) ›› 2016, Vol. 51 ›› Issue (5): 36-42.doi: 10.6040/j.issn.1671-9352.0.2015.609

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Purification, characterization and gene cloning of the extracellular protease EYHⅠ from Salinivibrio sp.YH4

WU Cui-ling1,2, LIU Dan1, YANG Xing-hao1, WU Ri-bang1, HUANG Jia-feng1, ZHANG Jiang1, LUN Zi-feng1, HE Hai-lun1*   

  1. 1.School of Life Sciences, State Key Laboratory of Medical Genetics, Central South University, Changsha 410013, Hunan, China;
    2. Department of Biochemistry, Changzhi Medical College, Changzhi 046000, Shanxi, China
  • Received:2015-12-17 Online:2016-05-20 Published:2016-05-16

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Abstract: The extracellular proteases from moderate halophilic strain, Salinivibrio sp. YH4 was isolated, purified, identified and characterized to provide data for enzyme system of Salinivibrio sp.. Strain was cultured in liquid fermentation medium, the extracellular proteases was purified by using HiTrap Capto DEAE column chromatography and Gel filtration chromatography. The protease was identified by using mass spectrometry(MS)and designated as EYHⅠ. The metalloprotease EYHⅠ had optimal activity at 55 ℃ and pH 9.0, stabilized at 60 ℃ and pH 7.0~11.0. EYHⅠshowed higher proteolytic activity at 4 mol/L NaCl. The nucleotide sequence contains 1836 bp, encoding EYHⅠof 611 amino acid residues, which is highly conserved to zinc metalloprotease precursor in Salinivibrio sp. AF-2004 with 96% sequence similarity. Analysis of the structure showed that EYHⅠcontained an FTP domain,a PepSY domain, M4 neutral protease and a PPC domain. These results may provide important data for the study of Salinivibrio sp. and the application of protease EYHⅠ.

Key words: purification, enzymic properties, gene cloning, Salinivibrio sp., proteases

CLC Number: 

  • Q936
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