Streptomyces, a genus of Gram-positive bacteria, are some of the most prolific antibiotic producers in the microbial world and are responsible for the production of over two-thirds of clinically relevant antibiotics. This makes them an important topic of study, especially in the wake of the fight against antimicrobial resistance (AMR). Understanding of the evolutionary forces at play in antibiotic synthesis could also be of use in the activation of so called ‘silent’ biosynthetic gene clusters (BGCs). To study the adaptation of Streptomyces coelicolor, a long-term evolution experiment (LTEE) was undertaken in the ‘superhost’ strain M1152, a strain without the major antibiotic BGCs, to allow the study of epistasis in relation to antibiotic biosynthesis. The LTEE consists of six distinct, liquid culture lineages, three grown in rich and three in minimal media. These cultures are passaged every three days (1:100 dilution) into fresh media with samples of each passage kept at -80°C for future analysis. Findings and observations from this experiment over 3000 generations include examples of parallel emergence of a fragmenting phenotype in multiple lineages, likely caused by a mutation in the matAB locus, adaptive diversification of carbon catabolism leading to a general increase in specific growth rate, and examples of negative epistasis in antibiotic production. We are now in the process of linking genotypes to phenotypes while further investigating the cause of epistasis including the putative role of a range of adpA alleles. These data highlight the importance of using experimental evolution in understanding antibiotic production and regulation