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Background  A bacterial artificial chromosomal library of  Planobispora rosea , a genetically intractable actinomycete strain, was constructed using  Escherichia coli - Streptomyces  artificial chromosome (ESAC) and screened for the presence of genes known to be involved in the biosynthesis of antibiotics.    Results  One clone with a 40 kb insert showed antimicrobial activity against Gram positive bacteria. Insert sequence analysis and subcloning experiments revealed that the bioactivity was due to a 3.5 kb DNA fragment containing two open reading frames. These  orfs  encode two proteins with high similarity to a putative membrane protein of  Streptomyces coelicolor  and to the nogalamycin resistance protein SnorO of  Streptomyces nogalater , respectively. The role of these two Orfs is unknown in  Planobispora.  Disruption and complementation experiments revealed that both proteins are necessary for the antibacterial activity and chemical analysis demonstrated that the antibiotic activity was due to thiostrepton, antibiotic used as recombinant clone selection marker.    Conclusion  Two  Planobispora rosea orfs  are responsible for increasing intracellular amounts and storage of thiostrepton in  Streptomyces lividans .

microbial cell factories

Two heterologously expressed Planobispora rosea proteins cooperatively induce Streptomyces lividans thiostrepton uptake and storage from the extracellular medium