Microbial production of geranyllinalool at near 10 g per liter

Abstract Background Acyclic diterpene alcohol, geranyllinalool (GL), is a natural insecticide and has diverse applications in cosmetics and pharmaceutical industries. The use of microbial cell factories for GL synthesis has been explored due to the non-sustainable nature of existing industrial manufacture methods. Results This study focused on three technical challenges encountered in GL cell factory construction in Saccharomyces cerevisiae , namely cellular supply of geranylgeranyl pyrophosphate, selection of GL synthases, and decisive factors for large-scale bioprocess development. With design for industrial uses in mind, a robust yet efficient terpenoids-synthesis enabling chassis strain was acquired. Next, a bifunctional prenyltransferase/terpene synthase enzyme (GLS12) of high efficiencies was identified and characterized in detail. Inclusion of GLS12 in the terpenoids chassis resulted in ~ 0.8 g/L of GL synthesis in shake-flask cultivations. Scale-up study was completed with ~ 9 g/L GL produced in a 10-L bioreactor in 96 h by fed-batch fermentation, the highest reported titer to date. Conclusions A high-performance yeast strain was engineered to enable efficient GL production using ethanol as the primary carbon source. This work demonstrated the potential of microbial GL fermentation for industrial uses.