Assessment of 4-nitro-1,8-naphthalic anhydride reductase activity in homogenates of bakers' yeast by reversed-phase high-performance liquid chromatography.
A simple reversed-phase high-performance liquid chromatographic (RP-HPLC) method was developed for the simultaneous determination of yield and conversion ratio of 4-nitro-1,8-naphthalic anhydride to 4-amino-1,8-naphthalic anhydride following incubation with a crude bakers' yeast homogenate. The analytes were separated on a C18 column in gradient mode. The detection limit of 4-amino-1,8-naphthalic anhydride is 10ng/microl when using a 10microl sample injection volume. The nitroreductase activity in the homogenate system can be assessed during the bioconversion process. The method can be used for the simultaneous detection of 4-hydroxylamino-1,8-naphthalic anhydride, an intermediate with limited stability.
Xu
Jianqiang
J
Department of Bioscience and Biotechnology, Dalian University of Technology, Dalian 116024, China.
Yang
Qing
Q
Qian
Xuhong
X
Samuelsson
Jörgen
J
Janson
Jan-Christer
JC
D000812:Anhydrides / Q000138:chemical synthesis / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D015394:Molecular Structure; D009601:Nitroreductases / Q000378:metabolism*; D015203:Reproducibility of Results; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D013056:Spectrophotometry, Ultraviolet
D000812:Anhydrides; D009601:Nitroreductases