| 초록 |
Biological production of limonene using microorganisms has been actively studied because it can replace hazardous organic solvents in chemical industry. In this study, we developed a bioconversion process from acetate and ethanol to limonene using Saccharomyces cerevisiae. Acetate and ethanol can be directly converted to acetyl-CoA not pyruvate providing, which has the advantage of solving the problems of carbon loss. To produce limonene from acetate and ethanol, we introduced limonene synthase(LS) gene from Mentha spicata which converts geranyl pyrophosphate(GPP) or neryl pyrophosphate(NPP) to limonene with GPP synthase(GPPS) from Abies grandis or NPP synthase(NDPS) from Solanum lycopersicum to S. cerevisiae using modified pESC-TRP vector. To improve the production of limonene, we optimized the mevalonate pathway by over-expression of acetyl-CoA thiolase, 3-hydroxy-3-methyl glutaryl-CoA(HMG-CoA) reductase, mevalonate kinase and IPP delta-isomerase which are the main bottle-neck of GPP synthesis using modified pESC-LEU and pESC-HIS vector. |
