Jasmonates play a pivotal role in multiple aspects of land plant physiology. On one hand, they are key drivers in the response to wounding and herbivore attack. On the other hand, they are crucial for plant development by facilitating flower development and fertility. Jasmonoyl-isoleucine (JA-Ile) is the bioactive jasmonate in vascular plants. It is formed through the activity of GRETCHEN HAGEN 3 (GH3) enzymes from the group of adenylating enzymes. The dicot model Arabidopsis thaliana harbors a total of 19 GH3 enzymes in its genome and so far important contributions to phytohormone-driven signaling for auxin, jasmonates and also SA have been described. Further analysis of this Brassicales specific clade of 11 GH3 enzymes from Arabidopsis revealed the discovery of novel substrate groups such as polyunsaturated fatty acids (PUFAs) and their oxidized derivatives as well as a pantothenic acid-phosphorylating enzyme activity. Together these data will be the basis for further studies on these novel enzyme activities.