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Abstract

The invention discloses a method of manufacturing a lysin protein capable of directly lysing Acinetobacter baumannii without pretreatment processing using chloroform or EDTA, comprising: transforming an expression plasmid into E. coli, wherein the expression plasmid is deposited at DSMA-Deutsche Sammlung von Mikroorganismen and Zellkulturen with deposit number DSM32023; expressing the expressing plasmid by the E. coli to form lysin protein having an amino acid sequence as set forth in SEQ ID NO: 6; lysing the E. coli containing the lysing protein to obtain a supernatant by centrifugating the E. coli lysate; mixing the supernatant and Ni²⁺ resins, washing the unbound protein by a binding buffer containing 50 mM Tris-HCl (pH 8.2), 15 mM MgCl2, 20% (v/v) glycerol, 0.05% β-ME and 0.1 mM PMSF; and eluting the purified lysing protein by an elution buffer containing 50 mM Tris-HCl (pH 8.2), 15 mM MgCl2, 20% (v/v) glycerol, 0.05% β-ME, 0.1 mM PMSF and 250 mM imidazole.

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First Claim