In summary, Corramycin features a novel antibiotic scaffold with unique or in natural products rarely observed structural features while displaying moderate to good anti-Gram-negative activity against numerous E. coli strains and S. typhimurium. Based on the unprecedented chemical structure of Corramycin and no observed cross-resistance with antibiotic classes commonly used in the clinic, we assume a novel, not yet elucidated, MoA. The identification of the biosynthetic pathway in C. coralloides underpins that myxobacterial species are a promising source of new antibiotic natural products with completely different chemistry compared to previously identified antibiotics, e.g., from actinobacteria. Furthermore, we proposed a biosynthesis model revealing interesting features such as a possible detoxification mechanism including the production of fatty acid-linked Corramycin precursors, which are exported and deacylated to secrete the active compound. Nevertheless, many proposed biosynthetic steps require further experimental investigations. Notably, the correct prediction of eight stereocenters, based on the in silico analysis of the assembly line, underlines the potential of bioinformatic tools in combination with chemical analysis methods in structure prediction of NRPS-derived natural products. We found a huge discrepancy between the FoR in rich medium compared to minimal medium due to the involvement of two transport systems in the Corramycin uptake. It is important to note that the typical development of an antibiotic would have been stopped with the high FoR found in the rich media. We here show that this process may be ineffective as the rich media typically employed in standardized FoR methods are certainly not good models for the infection process, because bacterial growth conditions are much less limited in rich media compared to minimal media, the latter mimicking in vivo conditions more realistically. Thus, antibiotics potentially useful for application in Corramycin serves as a prime example for such a scenario and our data clearly show the importance of more detailed studies using various media and of determining the potential mode(s) of resistance in the pathogen early in the drug discovery process. Taken into account the structural originality of the Corramycin scaffold, the possibility to prepare analogues by total synthesis, the anti-Gram-negative activity, the lack of cross-resistance with existing antibacterial classes suggesting a novel MoA, the unprecedented mode of penetration and its promising efficacy in a mouse septicemia model, we came to the conclusion that Corramycin constituted an attractive starting point for an antibacterial drug discovery program, despite some limitations (structural complexity, narrow spectrum, moderate potency and poor PK parameters). Subsequent efforts aim at preparing novel Corramycin analogues with more potent antimicrobial activities, a broadened spectrum against Gram-negative pathogens and acceptable pharmaceutical properties, with the objective to identify a preclinical development candidate. Given the unique structural and biological properties of Corramycins, we hope that this compound class will eventually deliver a marketable antibiotic.