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| Metabolic Engineering of Saccharomyces cerevisiae for Production of 8-Dimenthylally Naringenin |
| LI Bo1,2, LIANG Nan1,2, LIU Duo1,2, LIU Hong1,2, WANG Ying1,2, XIAO Wen-hai1,2, YAO Ming-dong1,2, YUAN Ying-jin1,2 |
1. Key Laboratory of Systems Bioengineering(Ministry of Education), Tianjin University, Tianjin 300072, China;
2. SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering, Tianjin 300072, China |
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Abstract 8-dimenthylallynaringenin(8DN) is the important precursor for the flavonoids medicine Icaritin, which has wide applications in pharmaceutical area. Due to the complexity of its biosynthetic pathway and related genes, main attentions were paied to biotransfering by procursor (like naringenin or isoxanthohumol) feeding. To date, no report about de novo biosynthesis of 8DN has been uncovered. In present study, in order to realize 8DN production in Saccharomyces cerevisiae, four key genes involved in naringenin biosynthesis, TAL,4CL,CHS and CHI from different species, were combinatorially screened, obtaining 30 naringenin producing strains, in which significant production variation ranged from 0.37mg/L to 22.34mg/L. Furthermore, the best combination of TAL,4CL,CHS and CHI genes were integrated into chromosome for better genetic stability by delta integration, generating strain SyBE_Sc02050031. Subsequently, prenyltransferase gene (N8DT) from Sophora flavescens incorporated with multicopy plasmid was introduced into SyBE_Sc02050031 (gaining strain SyBE_Sc02050032) and a titer of 8DN at 36.7μg/L in shaking flask was observed accordingly, indicating de novo biosynthesis of 8DN in yeast was successfully achieved. In addition, truncation tailoring strategy was explored to improve the catalysis function of N8DT, leading to 8DN production (up to 52.6μg/L) increasing by 44% compared to that in strain SyBE_Sc02050032. de novo biosynthesis of 8DN in microbes is firstly accomplished, which provides a good reference for microbial production of other natural flavonoids.
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Received: 15 March 2017
Published: 25 September 2017
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