Biosynthesis of pinene from glucose using metabolically-engineered Corynebacterium glutamicum

Pinene is a monoterpenes (C-10) that is produced in a genetically-engineered microbial host for its industrial applications in fragrances, flavoring agents, pharmaceuticals, and biofuels. Herein, we have metabolically-engineered Corynebacterium glutamicum, to produce pinene and studied its toxicity in C. glutamicum. Geranyl diphosphate synthases (GPPS) and pinene synthases (PS), obtained from Pinus taeda and Abies grandis, were co-expressed with over-expressed native 1-deoxy-d-xylulose-5-phosphate synthase (Dxs) and isopentenyl diphosphate isomerase (Idi) from C. glutamicum using CoryneBrick vector. Most strains expressing PS-GPPSs produced detectable amounts of pinene, but co-expression of DXS and IDI with PS (P. taeda) and GPPS (A. grandis) resulted in 27 mu g +/- A 7 alpha-pinene g(-1) cell dry weight, which is the first report in C. glutamicum. Further engineering of PS and GPPS in the C. glutamicum strain may increase pinene production.The authors thank Prof. Andreas Burkovski for providing the LN Delta GDH mutant and Ms. Jae Hee Jung for technical support. This work was supported by the National Research Foundation of Korea grant-funded by the Korean Government (Ministry of Science, ICT & Future Planning) (2014, University-Institute cooperation program) and Creative Allied Program (CAP) of the Korea Research Council of Fundamental Science and Technology (KRCF)/Korea Institute of Science and Technology (KIST) (Project No. 2E24832)