Provided herein is a novel homolog of Clostridial Botulinum neurotoxin (BoNT) identified in the bacterial species Enterococcus faecium (BoNT/EN). BoNT/EN variants and chimeric toxins comprising domains from BoNT/EN and other BoNTs are also provided. Further provided herein are methods of producing BoNT/EN and its use, e.g., in therapeutic applications.