Background:In recent years,food safety has become a global public health concern.Microbial contamination is one of the most common food safety issues.Staphylococcus aureus is a common foodborne pathogen that can form ...Background:In recent years,food safety has become a global public health concern.Microbial contamination is one of the most common food safety issues.Staphylococcus aureus is a common foodborne pathogen that can form biofilms on the surface of food processing equipment,leading to greater resistance to antimicrobial agents than occurs with planktonic bacteria.Materials and methods:In this work,recombinant Escherichia coli BL21(DE3)cells expressing optimized lysostaphin(Lst)were constructed,recombinant Lst was produced and purified,and Lst enzymatic assays were performed,followed by antimicrobial testing of Lst.Finally,a mixtureof Lst and DNase I was tested for antibiofilmactivity.Results:The protein content of purified Lst was 0.6 mg/mL and the enzyme activity was 240 U/mL.The minimum inhibitory concentration(MiC)of Lst against S.aureus was O.1μg/mL.At 1 MiC,Lst exerted an effect on the growth,cell wall integrity and cell membrane permeability of S.aureus.Conclusions:Although Lst alone also showed good inhibition and disruption of S.aureus biofilms,the inhibition and disruption of S.aureus biofilms were significantly greater when Lst was mixed with DNase l.This is probably because DNase I removes extracellular DNA,affecting biofilm formation and dispersing mature biofilms,and thereby facilitating the penetration of Lst.展开更多
基金financially supported by the Natural Science Foundation of Shandong Province Grant for Distinguished Young Scholars(No.ZR2022JQ15)the Young Taishan Scholars Program of Shandong Province(No.tsqn202103094)+1 种基金the Talent Research Foundation of Qingdao Agricultural University(No.665/1121020 and No.663/1117023)the Research Fund of Qingdao Special Food Research Institute(No.20220329),China.
文摘Background:In recent years,food safety has become a global public health concern.Microbial contamination is one of the most common food safety issues.Staphylococcus aureus is a common foodborne pathogen that can form biofilms on the surface of food processing equipment,leading to greater resistance to antimicrobial agents than occurs with planktonic bacteria.Materials and methods:In this work,recombinant Escherichia coli BL21(DE3)cells expressing optimized lysostaphin(Lst)were constructed,recombinant Lst was produced and purified,and Lst enzymatic assays were performed,followed by antimicrobial testing of Lst.Finally,a mixtureof Lst and DNase I was tested for antibiofilmactivity.Results:The protein content of purified Lst was 0.6 mg/mL and the enzyme activity was 240 U/mL.The minimum inhibitory concentration(MiC)of Lst against S.aureus was O.1μg/mL.At 1 MiC,Lst exerted an effect on the growth,cell wall integrity and cell membrane permeability of S.aureus.Conclusions:Although Lst alone also showed good inhibition and disruption of S.aureus biofilms,the inhibition and disruption of S.aureus biofilms were significantly greater when Lst was mixed with DNase l.This is probably because DNase I removes extracellular DNA,affecting biofilm formation and dispersing mature biofilms,and thereby facilitating the penetration of Lst.
