Nitrocefin is a chromogenic cephalosporin used to detect β-lactamases produced by β-lactam resistant bacteria. It does not appear to have antimicrobial properties, but has useful diagnostic properties. It can be used as a reagent in β-lactamase activity studies. Nitrocefin is soluble in DMSO.
||Nitrocefin is useful for the detection of β-lactamase patterns from bacterial cell extracts by isoelectric focusing. It can also be used in biosensor development and protein switch technology.
|Mechanism of Action
||β-lactamases hydrolyze the amide bond between the carbonyl carbon and the nitrogen in the β-lactam ring of Nitrocefin. Intact Nitrocefin is converted to hydrolyzed Nitrocefin under visible light (~ 380-500 nm) and a color change occurs from yellow (max at pH 7.0 = 390 nm) to red (max at pH 7.0 = 486 nm).
||Nitrocefin is used to detect β-lactamase activity from suspected β-lactam resistant bacteria (see Protocol). It is commonly used at a working concentration of 0.5 - 1.0 mg/ml.
Nitrocefin (TOKU-E) was used to study:
TEM-1 ß-lactamase activity:
"Label-free measurements of reaction kinetics using a droplet-based optofluidic device." Zhangming et al. See Link.
"Parts-per-million of polyethylene glycol as a non-interfering blocking agent for homogeneous biosensor development." Liu et al. See Link.
Surrogate ß-lactamase-nitrocefin assay:
"A cell-free fluorometric high-throughput screen for inhibitors of Rtt109-catalyzed histone acetylation." Dahlin et al. See Link.
"An unusual feruloyl esterase belonging to family VIII esterases and displaying a broad substrate range" Ohlhoff et al. See Link.
VIM-2 Metallo-ß-lactamases (MBLs):
"Inhibiting the VIM-2 Metallo-ß-Lactamase by graphene oxide and carbon nanotubes." Huang et al. See Link.
ß-lactamase production in E. coli and Klebsiella species:
"Occurrence of beta-lactamases and the antibiogram pattern of clinical isolates of Escherichia coli and Klebsiella species in Nsukka metropolis." Eze E et al. See Link.
Engineered protein switches:
"Electrochemical activation of engineered protein switches." Choi et al. See Link.
"Molecular determinants for protein stabilization by insertional fusion to a thermophilic host protein." Pierre et al. See Link.
Enzymatic protein switches:
"Enzymatic protein switches built from paralogous input domains." Tullman and Nicholes. See Link.
Guntas G, Mitchell S and Ostermeier M (2004) A molecular switch created by in vitro recombination of nonhomologous genes. Cell Press 11(11):1483-1487
O'Callaghan CH et al (1972) Novel method for detection of B-Lactamases by using a chromogenic cephalosporin substrate. Antimicrob. Agents. Chemother. 1(4):283-288
Parr, TR et al (1984) Simple screening method for beta-lactamase-positive and-negative Ampicillin-resistant Haemophilus Influenzae isolates. J. Clin. Microbiol. 20(1):131-132