Assay method for Escherichia coli photolyase activity using single-strand cis-syn cyclobutane pyrimidine dimer DNA as substrate.
A high-performance liquid chromatography method for the assay of Escherichia coli photolyase activity was developed. When cis-syn cyclobutane pyrimidine dimer was used as substrate, the Michaelis constant (K(m)) value for the photolyase activity was 100 nM. The linear range of the calibration curve of the photolyase activity was 0.026-6.64 microU/assay tube. The correlation coefficient for this linearity was 0.998. The limit of detection (S/N = 3) was 26 nU/assay tube. The photolyase activity was increased 1.6-fold in the presence of 5,10-methenyltetrahydrofolic acid in the enzyme reaction mixture.
Nakayama
Takuya
T
Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
Todo
Takeshi
T
Notsu
Saori
S
Nakazono
Manabu
M
Zaitsu
Kiyoshi
K
D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004247:DNA; D004255:Deoxyribodipyrimidine Photo-Lyase / Q000032:analysis* / Q000187:drug effects; D004926:Escherichia coli / Q000187:drug effects / Q000201:enzymology*; D007700:Kinetics; D011622:Pterins / Q000494:pharmacology; D011740:Pyrimidine Dimers / Q000138:chemical synthesis; D014466:Ultraviolet Rays
D011622:Pterins; D011740:Pyrimidine Dimers; D004247:DNA; D004255:Deoxyribodipyrimidine Photo-Lyase