<jats:title>Abstract</jats:title> <jats:p>Sperabillins, novel antibiotics, were isolated from the culture filtrate of Pseudomonas fluorescens YK-437. The structure of sperabillin A was elucidated to be 3-[[(3R,5R)-3-amino-6-[(2E,4Z)-2,4-hexadienoylamino]-5-hydroxyhexanoyl]amino]propanamidine dihydrochloride. Sperabillin B has a methyl substituent at the C-6 position of sperabillin A. Sperabillin C and D are (2E,4E)-isomers of the 2,4-hexadienoyl moiety of sperabillin A and B, respectively.