2-Nitro-5-(6-bromohexanoylamino)benzoic acid test paper method for detecting microorganisms capable of producing cephalosporin acylases.
A novel method for detecting microorganisms capable of producing cephalosporin C (CPC) acylase and/or 7-(4-carboxybutanamido)cephalosporanic acid (GL-7-ACA) acylase has been developed. The method is based on the degradation of 2-nitro-5-(6-bromohexanoylamino)benzoic acid (NBHAB), a chromogenic substrate, into yellow 2-nitro-5-aminobenzoic acid by the action of the CPC acylase or the GL-7-ACA acylase. This method is very sensitive and quite specific, and has been successfully applied to screen the acylases from a variety of bacteria. A large number of colonies isolated on a plate surface from more than 67 samples and several known bacteria were tested by the NBHAB paper. Five NBHAB-positive strains and isolates were obtained. They were further examined by the reaction of their bacterial cells upon CPC and GL-7-ACA, respectively, and by thin-layer chromatography in order to distinguish the CPC acylase from the GL-7-ACA acylase.
Zhang
Q J
QJ
Institute of Biophysics, Chinese Academy of Sciences, Beijing.
Xu
W X
WX
Shi
L
L
D000362:Agar; D000581:Amidohydrolases / Q000032:analysis; D001407:Bacillus / Q000201:enzymology*; D004926:Escherichia coli / Q000201:enzymology*; D009579:Nitrobenzoates; D010399:Penicillin Amidase; D011549:Pseudomonas / Q000201:enzymology*; D011934:Reagent Strips; D013379:Substrate Specificity
D009579:Nitrobenzoates; D011934:Reagent Strips; D000362:Agar; C072509:2-nitro-5-(6-bromohexanoylamino)benzoic acid; D000581:Amidohydrolases; D010399:Penicillin Amidase; C046586:glutarylamidocephalosporanic acid acylase