Development of antibiotic-overproducing strains by site-directed mutagenesis of the rpsL gene in Streptomyces lividans.
Certain rpsL (which encodes the ribosomal protein S12) mutations that confer resistance to streptomycin markedly activate the production of antibiotics in Streptomyces spp. These rpsL mutations are known to be located in the two conserved regions within the S12 protein. To understand the roles of these two regions in the activation of silent genes, we used site-directed mutagenesis to generate eight novel mutations in addition to an already known (K88E) mutation that is capable of activating antibiotic production in Streptomyces lividans. Of these mutants, two (L90K and R94G) activated antibiotic production much more than the K88E mutant. Neither the L90K nor the R94G mutation conferred an increase in the level of resistance to streptomycin and paromomycin. Our results demonstrate the efficacy of the site-directed mutagenesis technique for strain improvement.
Okamoto-Hosoya
Yoshiko
Y
National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan.
Okamoto
Susumu
S
Ochi
Kozo
K
D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000494:pharmacology; D024881:Drug Resistance, Bacterial; D005818:Genetic Engineering / Q000379:methods; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010303:Paromomycin / Q000096:biosynthesis* / Q000494:pharmacology; D010957:Plasmids; D012269:Ribosomal Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D013302:Streptomyces / Q000235:genetics / Q000378:metabolism*; D013307:Streptomycin / Q000096:biosynthesis* / Q000494:pharmacology
D000900:Anti-Bacterial Agents; D012269:Ribosomal Proteins; C034212:ribosomal protein S12; D010303:Paromomycin; D013307:Streptomycin