Jasmonates are a class of plant hormones with many agricultural applications and potential medicinal properties. However, the low content of jasmonates in plants and environmental issues with their production make their supply challenging. In the present study, we report the de novo microbial biosynthesis of jasmonic acid and its derivatives, methyl jasmonate and jasmonoyl isoleucine, from glucose using an engineered baker’s yeast. The study uses enzymes located in the endoplasmic reticulum and cytosol to generate the intermediates α-linolenic acid and cis-12-oxophytodienoic acid. Our final engineered stain, which integrates 15 heterologous genes from diverse plants and fungi and had 3 of its native genes deleted, produces jasmonic acid at titres of 19.0 mg l−1 in flask cultures through in vitro supplementation of α-linolenic acid. In addition to the well-known natural structures (−)-jasmonic acid and (+)-epi-jasmonic acid, the engineered yeast also synthesized the previously unobserved unnatural structures (+)-jasmonic acid and (−)-epi-jasmonic acid. These results demonstrate that yeast is a scalable and sustainable platform to produce both naturally occurring jasmonates and those structures not found naturally in plants. (Figure presented.)