To produce a library of S. fungicidicus mutants, an in vitro transposon-based method was devised to randomly insert a selectable marker and oriT element in fosmid inserts carrying segments of the enramycin biosynthetic gene cluster and adjacent regions. This library of mutagenized fosmids was screened to identify specific disrupted genes. The collection of mutagenized fosmids was then introduced to the parent wild-type Streptomyces sp. to create a cluster-wide disruption library. Advantageous mutations were identified leading to organisms that biosynthesize different enduracidin analogs. Exemplary enduracidin analogs synthesized by this method include tetrahydrorenduracidin A, tetrahydroenduracidin B, deschloroenduracidin A, and deschloroenduracidin B.
Features & Benefits
Background of Invention
The emerging and persistent threat of multidrug-resistant bacterial infections has resulted in enormous healthcare costs and poses major challenges to public health. To address the need for new antibiotics capable of combating the gowning problem of resistance, Oregon State University researchers cloned and patented the biosynthetic gene cluster for Enramycin, which is a peptide antibiotic produced in S. fungicidicus and widely used as a feed additive for pigs and chickens, and also developed methods for generating a library of enramycin analogs.
U.S. patent no. 8,188,245 ; available for exclusive or non-exclusive licensing in a field limited to human health and therapeutics.