Aim: This study focuses on the production, purification and characterization of serraticin A, a novel cold-active antimicrobial produced by Serratia proteamaculans 136. Methods and Results: A Ser. proteamaculans strain producing a novel cold-active antimicrobial was isolated from Isla de los Estados, Argentina. Antimicrobial production was optimized in a BIOFLO 101 bioreactor under batch culture mode, with temperature, pH and dissolved oxygen controlled conditions. A purification protocol was developed including activated charcoal adsorption, solid-phase C18 extraction (SPE) and semi-preparative HPLC. The molecular weight was determined by LC/QTOF/MS/MS mass analysis. Conclusions: Serratia proteamaculans 136 produces a cold-active low molecular bacteriocin-like compound named serraticin A. In this work, it has been laboratory-scale produced, purified and partially characterized. Cross-immunity test revealed that serraticin A is very different from other well-known microcins assayed, with a wide inhibitory spectrum, showing an interesting biotechnology potential to be applied as a control agent against pathogenic bacteria. Significance and Impact of the Study: The present study is the first report of a cold-active compound with antimicrobial activity from Ser. proteamaculans. The work also highlights that cold environments could be a suitable source of micro-organisms with ability to produce cold-active biomolecules of biotechnological interest.