PMID: 2120199Oct 1, 1990Paper

Functional expression of 8-hydroxy-5-deazaflavin-dependent DNA photolyase from Anacystis nidulans in Streptomyces coelicolor

Journal of Bacteriology
F MayerlA Yasui

Abstract

The gene encoding Anacystis nidulans 5-deazaflavin-dependent photolyase (phr) was inserted into the Streptomyces vector pIJ385 to form a transcriptional fusion with the neomycin resistance (aph) gene. The resulting plasmid, pANPL, was introduced into Streptomyces coelicolor, a host which exhibits no detectable photolyase activity and provides 5-deazaflavins. Transformants expressed functional photolyase and could be cultured at much higher cell densities than A. nidulans. A two-step affinity protocol was used to purify photolyase to homogeneity. High-pressure liquid chromatographic analysis established the presence of 5-deazaflavin cofactors in the enzyme, showing that this expression system allows heterologous production of 5-deazaflavin-class photolyases.

References

Jun 26, 1989·Nucleic Acids Research·T KobayashiA Yasui
Jul 1, 1989·Chemical Research in Toxicology·G M Myles, A Sancar
Apr 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·J L JohnsonA Sancar
Jan 1, 1988·Annual Review of Biochemistry·A Sancar, G B Sancar
Jun 1, 1987·Journal of Bacteriology·I Husain, A Sancar
May 26, 1987·Nucleic Acids Research·D Pulido, A Jiménez
Jan 1, 1986·Methods in Enzymology·R P HausingerC Walsh
Sep 1, 1983·Proceedings of the National Academy of Sciences of the United States of America·C J Thompson, G S Gray
Jan 15, 1984·Journal of Molecular Biology·A Sancar, G B Sancar

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