Group B Streptococci (GBS) are β-hemolytic Gram-positive bacteria frequently associated with fetal injury, preterm birth, spontaneous abortion, and neonatal infection. GBS disease remains a major global health problem, as current antibiotic-based prevention strategies have limitations. In order to develop improved, rationally designed prevention strategies for GBS infection and disease, a comprehensive understanding of GBS virulence factors is essential. A key virulence factor for GBS is the unique pigmented ornithine rhamnolipid, which confers hemolysis and induces cytotoxicity in a number of host cells, including neutrophils, monocytes, macrophages, and amniotic cells. Here, we add to the body of knowledge about the GBS hemolytic pigment toxin by showing that heterologous expression of the GBS cyl operon in non-hemolytic Lactococcus lactis results in hemolysis, pigmentation, and cytotoxicity to neutrophils. We isolated the hemolytic pigment toxin from cyl-complemented L. lactis and performed NMR analysis and hemolytic assays, which confirmed the presence of the active hemolytic ornithine rhamnolipid in this strain. Together, these findings provide evidence that that the cyl operon is sufficient for the production of the active GBS hemolytic pigment toxin in a non-pathogenic Gram-positive bacterial strain. Further studies are in progress and will reveal new information about this unique toxin.