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 |
2007-7-1
|
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 |
2.366
|
Times Cited |
5
|
WOS Category
|
FOOD SCIENCE & TECHNOLOGY
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
|
Resource |
General Microbes |
JCM 1028
JCM 1055
JCM 9703 |