Biosynthesis of Chlorinated Lactylates in <i>Sphaerospermopsis</i> sp. LEGE 00249

Lactylates are an important group of molecules in the food and cosmetic industries. A series of natural halogenated 1-lactylates, chlorosphaerolactylates (<b>1</b>–<b>4</b>), were recently reported from <i>Sphaerospermopsis</i> sp. LEGE 00249. Here, we identify the <i>cly</i> biosynthetic gene cluster, containing all the necessary functionalities for the biosynthesis of the natural lactylates, based on in silico analyses. Using a combination of stable isotope incorporation experiments and bioinformatic analysis, we propose that dodecanoic acid and pyruvate are the key building blocks in the biosynthesis of <b>1</b>–<b>4</b>. We additionally report minor analogues of these molecules with varying alkyl chains. This work paves the way to accessing industrially relevant lactylates through pathway engineering.