PMID:15158485

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Citation

Nakayama, T, Todo, T, Notsu, S, Nakazono, M and Zaitsu, K (2004) Assay method for Escherichia coli photolyase activity using single-strand cis-syn cyclobutane pyrimidine dimer DNA as substrate. Anal. Biochem. 329:263-8

Abstract

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.

Links

PubMed Online version:10.1016/j.ab.2004.03.021

Keywords

Chromatography, High Pressure Liquid/methods; DNA; Deoxyribodipyrimidine Photo-Lyase/analysis; Deoxyribodipyrimidine Photo-Lyase/drug effects; Escherichia coli/drug effects; Escherichia coli/enzymology; Kinetics; Pterins/pharmacology; Pyrimidine Dimers/chemical synthesis; Ultraviolet Rays

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