RRC ID 15443
Author Ahmed AB, Noguchi K, Asami Y, Nomura K, Fujii H, Sakata M, Tokita A, Noda K, Kuroda A.
Title Evaluation of cell wall binding domain of Staphylococcus aureus autolysin as affinity reagent for bacteria and its application to bacterial detection.
Journal J. Biosci. Bioeng.
Abstract We evaluated the cell wall binding (CWB) domain of Staphylococcus aureus autolysin as an affinity reagent for bacteria. A fusion of CWB domain and green fluorescent protein (CWB-GFP) bound to S. aureus with a dissociation constant of 15 nM. CWB-GFP bound to a wide range of gram-positive bacteria, but not to most gram-negative bacteria. We suspected that the outer membrane of gram-negative bacteria inhibits the access of CWB-GFP to peptidoglycan layer. Indeed, CWB-GFP bound to gram-negative bacteria when they were treated with benzalkonium chloride. Because CWB-GFP bound to the bacterial peptidoglycan layer, it appeared to be an effective affinity reagent for bacteria and CWB fusion with reporter proteins could be applied to detect bacteria. We also constructed a fusion of CWB and luciferase, which can be used for the rapid detection of bacteria.
Volume 104(1)
Pages 55-61
Published 2007-7
DOI 10.1263/jbb.104.55
PII S1389-1723(07)70119-6
PMID 17697984
MeSH Bacteria / chemistry Bacteria / isolation & purification* Bacteriological Techniques / methods* Cell Wall / chemistry* Green Fluorescent Proteins / analysis* Green Fluorescent Proteins / genetics Lactobacillus / chemistry Lactobacillus / isolation & purification Luciferases / analysis Luciferases / genetics N-Acetylmuramoyl-L-alanine Amidase / analysis N-Acetylmuramoyl-L-alanine Amidase / chemistry* N-Acetylmuramoyl-L-alanine Amidase / genetics Protein Structure, Tertiary Recombinant Fusion Proteins / analysis Recombinant Fusion Proteins / chemistry Recombinant Fusion Proteins / genetics Spores, Bacterial / chemistry Spores, Bacterial / isolation & purification Staphylococcus aureus / enzymology*
IF 2.015
Times Cited 3
General Microbes JCM 1028 JCM 1055 JCM 9703