Open AccessFunctional expression of 8-hydroxy-5-deazaflavin-dependent DNA photolyase from Anacystis nidulans in Streptomyces coelicolor
Author(s) -
Friedrich Mayerl,
Jacqueline Piret,
Andreas Kiener,
Charles J. Walsh,
Akira Yasui
Publication year - 1990
Publication title -
journal of bacteriology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.652
H-Index - 246
eISSN - 1067-8832
pISSN - 0021-9193
DOI - 10.1128/jb.172.10.6061-6065.1990
Subject(s) - photolyase , biology , aspergillus nidulans , streptomyces coelicolor , streptomyces albus , streptomyces , plasmid , heterologous , thiostrepton , gene , microbiology and biotechnology , biochemistry , genetics , dna repair , bacteria , mutant , rna , ribosome
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.