Assay method for Escherichia coli photolyase activity using single-strand cis-syn cyclobutane pyrimidine dimer DNA as substrate

Takuya Nakayama, Takeshi Todo, Saori Notsu, Manabu Nakazono, Kiyoshi Zaitsu

Research output: Contribution to journalArticle

6 Citations (Scopus)

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 (Km) value for the photolyase activity was 100nM. The linear range of the calibration curve of the photolyase activity was 0.026-6.64μU/assay tube. The correlation coefficient for this linearity was 0.998. The limit of detection (S/N=3) was 26nU/assay tube. The photolyase activity was increased 1.6-fold in the presence of 5,10-methenyltetrahydrofolic acid in the enzyme reaction mixture.

Original languageEnglish
Pages (from-to)263-268
Number of pages6
JournalAnalytical Biochemistry
Volume329
Issue number2
DOIs
Publication statusPublished - Jun 15 2004

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Deoxyribodipyrimidine Photo-Lyase
Pyrimidine Dimers
Escherichia coli
Assays
DNA
Substrates
High performance liquid chromatography
Calibration
Limit of Detection
High Pressure Liquid Chromatography
Acids
Enzymes

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Assay method for Escherichia coli photolyase activity using single-strand cis-syn cyclobutane pyrimidine dimer DNA as substrate. / Nakayama, Takuya; Todo, Takeshi; Notsu, Saori; Nakazono, Manabu; Zaitsu, Kiyoshi.

In: Analytical Biochemistry, Vol. 329, No. 2, 15.06.2004, p. 263-268.

Research output: Contribution to journalArticle

Nakayama, Takuya ; Todo, Takeshi ; Notsu, Saori ; Nakazono, Manabu ; Zaitsu, Kiyoshi. / Assay method for Escherichia coli photolyase activity using single-strand cis-syn cyclobutane pyrimidine dimer DNA as substrate. In: Analytical Biochemistry. 2004 ; Vol. 329, No. 2. pp. 263-268.
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