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Attenuating the Taxol yield of  Aspergillus terreus  with the subculturing and storage were the technical challenges that prevent this fungus to be a novel platform for industrial Taxol production. Thus, the objective of this study was to unravel the metabolic machineries of  A .  terreus  associated with attenuation of Taxol productivity, and their restoring potency upon cocultivation with the  Podocarpus gracilior  microbiome. The Taxol yield of  A .  terreus  was drastically reduced with the fungal subculturing. At the 10 th  subculture, the yield of Taxol was reduced by four folds (78.2 µg/l) comparing to the original culture (268 µg/l), as authenticated from silencing of molecular expression of the Taxol-rate limiting enzymes (GGPPS, TDS, DBAT and BAPT) by qPCR analyses. The visual fading of  A .  terreus  conidial pigmentation with the subculturing, revealing the biosynthetic correlation of melanin and Taxol. The level of intracellular acetyl-CoA influx was reduced sequentially with the fungal subculturing, rationalizing the decreasing on Taxol and melanin yields. Fascinatingly, the Taxol biosynthetic machinery and cellular acetyl-CoA of  A .  terreus  have been completely restored upon addition of 3% surface sterilized leaves of  P .  gracilior , suggesting the implantation of plant microbiome on re-triggering the molecular machinery of Taxol biosynthesis, their transcriptional factors, and/or increasing the influx of Acetyl-CoA. The expression of the proteins of 74.4, 68.2, 37.1 kDa were exponentially suppressed with  A .  terreus  subculturing, and strongly restored upon addition of  P .  gracilior  leaves, ensuring their profoundly correlation with the molecular expression of Taxol biosynthetic genes. From the proteomic analysis, the restored proteins 74.4 kDa of  A .  terreus  upon addition of  P .  gracilior  leaves were annotated as ribosome biogenesis proteins YTM and microtubule-assembly proteins that belong to WD40 superfamily. Thus, further ongoing studies for molecular cloning and expression of these genes with strong promotors in  A .  terreus , have been initiated, to construct a novel platform of metabolically stable  A .  terreus  for sustainable Taxol production. Attenuating the Taxol yield of  A .  terreus  with the multiple-culturing and storage might be due to the reduction on main influx of acetyl-CoA, or downregulation of ribosome biogenesis proteins that belong to WD40 protein superfamily.

scientific reports

Restoring the Taxol biosynthetic machinery of Aspergillus terreus by Podocarpus gracilior Pilger microbiome, with retrieving the ribosome biogenesis proteins of WD40 superfamily