Synthesis of 2-arylbenzothiazole derivatives and their application in bacterial detection.
A series of 2-arylbenzothiazole derivatives have been prepared as fluorogenic enzyme substrates in order to detect aminopeptidase, esterase, phosphatase and β-galactosidase activity in clinically important Gram-negative and Gram-positive bacteria. Substrates were incorporated into an agar-based culture medium and this allowed growth of intensely fluorescent bacterial colonies based on hydrolysis by specific enzymes. Substrate 20 targeted L-alanine aminopeptidase activity and was hydrolysed exclusively by a range of Gram-negative bacteria and inhibited the growth of a range of Gram-positive bacteria. Substrate 19a targeted β-alanyl aminopeptidase activity and generated fluorescent colonies of selected Gram-negative species including Pseudomonas aeruginosa. Substrate 21b targeted C8-esterase activity and resulted in strongly fluorescent colonies of selected species known to harbour such enzyme activity (e.g., Salmonella and Pseudomonas). Most Gram-negative species produced colonies with an intense blue fluorescence due to hydrolysis of phosphatase substrates 24a-c and substrate 24c was also hydrolysed by strains of Staphylococcus aureus. Compounds 26b and 26c targeted β-galactosidase activity and generated strongly fluorescent colonies with coliform bacteria that produced this enzyme (e.g., Escherichia coli).
Research & Development Microbiology, bioMérieux SA, 3 route de Port Michaud, 38 390 La-Balme-les-Grottes, France.
Marie
M
Cellier
Department of Applied Sciences, Northumbria University, Newcastle upon Tyne NE1 8ST, UK.
Elizabeth
E
Fazackerley
Department of Applied Sciences, Northumbria University, Newcastle upon Tyne NE1 8ST, UK.
Arthur L
AL
James
Research & Development Microbiology, bioMérieux SA, 3 route de Port Michaud, 38 390 La-Balme-les-Grottes, France.
Sylvain
S
Orenga
Department of Microbiology, Freeman Hospital, Newcastle upon Tyne NE7 7DN, UK.
John D
JD
Perry
Department of Applied Sciences, Northumbria University, Newcastle upon Tyne NE1 8ST, UK.
Graeme
G
Turnbull
Department of Applied Sciences, Northumbria University, Newcastle upon Tyne NE1 8ST, UK. Electronic address: steven.stanforth@northumbria.ac.uk.
Stephen P
SP
Stanforth
D000626:Aminopeptidases / Q000378:metabolism; D052160:Benzothiazoles / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D004950:Esterases / Q000378:metabolism; D005456:Fluorescent Dyes / Q000138:chemical synthesis* / Q000378:metabolism / Q000494:pharmacology; D006090:Gram-Negative Bacteria / Q000187:drug effects / Q000201:enzymology / Q000302:isolation & purification; D006094:Gram-Positive Bacteria / Q000187:drug effects / Q000201:enzymology / Q000302:isolation & purification; D006868:Hydrolysis; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D013379:Substrate Specificity; D001616:beta-Galactosidase / Q000378:metabolism
D052160:Benzothiazoles; D005456:Fluorescent Dyes; D004950:Esterases; D010744:Phosphoric Monoester Hydrolases; D001616:beta-Galactosidase; D000626:Aminopeptidases; C005465:benzothiazole
Aminopeptidase; Bacteria detection; Esterase; Fluorogenic substrates; Phosphatase; β-Galactosidase