Sustainable Production of Bio-Based Geraniol: Heterologous Expression of Early Terpenoid Pathway Enzymes in Chlamydomonas reinhardtii

Geraniol is a monoterpene alcohol with a rose-like aroma, used in food and cosmetics and for its anti-inflammatory, antibacterial, and insect-repellent properties. Geraniol is commonly chemically synthesized from petroleum-based sources in a highly energy-demanding process with a large carbon footprint. Alternatively, geraniol can be derived from plant-based essential oils but with relatively low yields and limitations from seasonal cultivation. Here, a sustainable geraniol biosynthesis alternative was established in the photosynthetic green microalga Chlamydomonas reinhardtii. Three enzymesgeraniol synthase from Catharanthus roseus (CrGES), geranyl diphosphate synthase from Lithospermum erythrorhizon (LeGPPS), and a modified 1-deoxy-d-xylulose-5-phosphate synthase from Salvia pomifera (SpDXS)were strategically redesigned for high expression from the algal nuclear genome. Various enzyme combinations and subcellular localizations were tested, resulting after 48 h in up to 1 mg geraniol/L (corresponding to 1.8 mg/g of dry weight) secreted into the culture medium. This work demonstrates a promising route for sustainable, CO2-based production of geraniol in microalgae and provides a foundation for further optimization.