Stimulation of Actinorhodin Production by Streptomyces lividans with Chromosomally-Integrated Antibiotic Regulatory Gene, afsR2

Streptomyces lividans is one of the most commonly-used streptomyetes strain as a molecular cloning and expression host. Unlike its close relative S. coelicolor, however, S. lividans rarely produces secondary metabolite such as actinorhodin in a typical glucose-containing culture condition due to insufficient expression of some antibiotic regulatory genes including afsR2. Although multiple copies of afsR2 or a glycerol-specific culture condition stimulated actinorhodin production in S. lividans, both failed to stimulate actinorhodin production in S. lividans cultured in a typical glucose-containing medium. To generate a culture-condition-independent actinorhodin-overproducing S. lividans strain the afsR2 gene was integrated into the S. lividans TK21 chromosome via homologous recombination, followed by the genetic confirmation. This S. lividans strain produced a significant amount of actinorhodin in both glucose-containing liquid and plate cultures, with higher actinorhodin productivity compared to the S. lividans containing multiple copies of afsR2. These results suggest that a chromosomal integration of a single copy of an antibiotic regulatory gene is a promising method for the development of a stable antibiotic-overproducing streptomycetes strain.