Design and Assembly of a Biofactory for (2)-Naringenin Production in : Effects of Oxygen Transfer on Yield and Gene Expression.

The molecule (2)-naringenin is a scaffold molecule with several nutraceutical properties. Currently, (2)-naringenin is obtained through chemical synthesis and plant isolation. However, these methods have several drawbacks. Thus, heterologous biosynthesis has emerged as a viable alternative to its production. Recently, (2)-naringenin production studies in have used different tools to increase its yield up to 588 mg/L. In this study, we designed and assembled a bio-factory for (2)-naringenin production. Firstly, we used several parametrized algorithms to identify the shortest pathway for producing (2)-naringenin in , selecting the genes phenylalanine ammonia lipase (), 4-coumarate: CoA ligase (), chalcone synthase (), and chalcone isomerase () for the biosynthetic pathway. Then, we evaluated the effect of oxygen transfer on the production of (2)-naringenin at flask (50 mL) and bench (4 L culture) scales. At the flask scale, the agitation rate varied between 50 rpm and 250 rpm. At the bench scale, the dissolved oxygen was kept constant at 5% DO (dissolved oxygen) and 40% DO, obtaining the highest (2)-naringenin titer (3.11 ± 0.14 g/L). Using genome-scale modeling, gene expression analysis (RT-qPCR) of oxygen-sensitive genes was obtained.