Heterologous microbial biocynthesis of saffron apocarotenoids using zeaxanthin previous substance

Plant secondary metabolites; color of flowers, taste of food, resistance to pests and diseases is very important for such features. Moreover, medicines are the source of many valuable chemicals such as dyes and odors. Therefore, the production of secondary metabolites in plants, producing a better chemical, producing a less toxic compound or being able to engineer to produce new compounds has attracted considerable interest. Studies on plant secondary metabolites have increased in recent years. New areas of development of the production of secondary metabolites have begun to emerge. Emerging areas, such as recombinant DNA technology, have aimed to improve secondary metabolite production by studying secondary metabolite pathways. However, some problems on these pathways have not been overcome. In order to overcome these shortcomings, new and solution oriented studies such as metabolic engineering and synthetic biology have brought success. By using microorganisms such as bacteria and yeasts, it was tried to increase the metabolite production by integrating the pathway structure of the metabolites produced in plants. In this study, pathway genes were transferred to bacteria without the need for plant and plant secondary metabolites production stages were developed. Secondary metabolites found in saffron (Crocus sativus L.), crocin, safranal and picrocrocin synthesis stages were investigated. Plasmid constructs of CCD2 and UGT genes were performed for metabolite biosynthesis. E. coli NEB 10-beta and E. coli MG1655 strains were used for expression of the plasmid constructs. Biosynthesis of saffron metabolites was investigated using HPLC. As a result of the studies, it was observed that picrocrocin production was observed on the basis of HPLC analysis but safranal and crocin biosynthesis was not realized. Alternative and interesting studies have been conducted in order to produce plant-independent saffron plant which has economic value in many fields. Key words: Secondary metabolite, synthetic biology, metabolic engineering, saffron, crocin, picrocrocin, saffranal, Crocus sativus L.