Expression and Antibacterial Activity of CHAP Catalytic Domain of Staphylococcus aureus Phage Lysin Ply187

doi:10.13560/j.cnki.biotech.bull.1985.2016.09.031

Abstract

Abstract: Bacteriophage lysins are efficiently antibacterial proteins,thus systematically analysing the activity of catalytic domain of it is conducive to the design of efficient heterozygous lysins. The coding sequence of the CHAP catalytic domain of Ply187（CHAP_Ply187）was synthesized,and a recombinant expression plasmid pET28a-CHAP_Ply187was constructed,which then was transformed into Escherichia coli BL21（DE3）. The highly pure recombinant CHAP_Ply187 protein was produced by IPTG induction,further purified by a two-step method,reaching > 95% purity,and finally compared with the catalytic domain of lysostaphin（CAT_Lysn）. The results from turbidimetry showed that both CHAP_Ply187 and CAT_Lysn possessed the solid antibacterial activities to Staphylococcus aureus.CHAP_Ply187showed a much broader antibacterial spectrum and optimal activity in a wider range of the pH,tolerated higher ionic strength,and more easily affected by EDTA. Ca²⁺,Mg²⁺,and Mn²⁺in low concentration significantly promoted the catalytic domains of both proteins,while the Zn²⁺,Cu²⁺,and Fe²⁺in low concentration strongly inhibited the catalytic domains.

Key words: bacteriophage lysin, lysostaphin, catalytic domain, CHAP, antibacterial activity

WU Meng¹, LU Hai-rong^{1, 2}, HUANG Qing-shan^{1, 2}. Expression and Antibacterial Activity of CHAP Catalytic Domain of Staphylococcus aureus Phage Lysin Ply187[J]. Biotechnology Bulletin, 2016, 32(9): 232-238.

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