Synergistic Regulation of Metabolism by Ca 2+ /Reactive Oxygen Species in Penicillium brevicompactum Improves Production of Mycophenolic Acid and Investigation of the Ca 2+ Channel

Although is a very important industrial strain for mycophenolic acid production, there are no reports on Ca /reactive oxygen species (ROS) synergistic regulation and calcium channels, Cch-pb. This study initially intensified the concentration of the intracellular Ca in the high yielding mycophenolic acid producing strain NRRL864 to explore the physiological role of intracellular redox state in metabolic regulation by . The addition of Ca in the media caused an increase of intracellular Ca , which was accompanied by a strong increase, 1.5 times, in the higher intracellular ROS concentration. In addition, the more intensive ROS sparked the production of an unreported pigment and increase in mycophenolic acid production. Furthermore, the Ca channel, the homologous gene of Cch1, Cch-pb, was investigated to verify the relationship between Ca and the intracellular ROS. The hemoglobin was overexpressed, which was bacterial hemoglobin from , reducing the intracellular ROS concentration to verify the relationship between the redox state and the yield of mycophenolic acid. The strain expressed the hemoglobin exhibited a lower intracellular ROS concentration, 30% lower, and decreased the yield of mycophenolic acid as 10% lower at the same time. Subsequently, with the NRRL864 fermented under 1.7 and 28 mM Ca , the [NADH]/[NAD ] ratios were detected and the higher [NADH]/[NAD ] ratios (4 times higher with 28 mM) meant a more robust primary metabolism which provided more precursors to produce the pigment and the mycophenolic acid. Finally, the 10 times higher calcium addition in the media resulted in 25% enhanced mycophenolic acid production to 6.7 g/L and induced pigment synthesis in NRRL864.