Identification and Functional Analysis of the Mycophenolic Acid Gene Cluster of Penicillium roqueforti

Abstract

The filamentous fungus Penicillium roqueforti is widely known as the ripening agent of blue-veined cheeses. Additionally, this fungus is able to produce several secondary metabolites, including the meroterpenoid compound mycophenolic acid (MPA). Cheeses ripened with P. roqueforti are usually contaminated with MPA. On the other hand, MPA is a commercially valuable immunosuppressant. However, to date the molecular basis of the production of MPA by P. roqueforti is still unknown. Using a bioinformatic approach, we have identified a genomic region of approximately 24.4 kbp containing a seven-gene cluster that may be involved in the MPA biosynthesis in P. roqueforti. Gene silencing of each of these seven genes (named mpaA, mpaB, mpaC, mpaDE, mpaF, mpaG and mpaH) resulted in dramatic reductions in MPA production, confirming that all of these genes are involved in the biosynthesis of the compound. Interestingly, the mpaF gene, originally described in P. brevicompactum as a MPA self-resistance gene, also exerts the same function in P. roqueforti, suggesting that this gene has a dual function in MPA metabolism. The knowledge of the biosynthetic pathway of MPA in P. roqueforti will be important for the future control of MPA contamination in cheeses and the improvement of MPA production for commercial purposes.

Identification of the pyranonigrin A biosynthetic gene cluster by genome mining in Penicillium thymicola IBT 5891

“…This is similar to the genomes of mycophenolic acid (MPA)‐producing strains . MPA is a known inhibitor of IMPDH, and previous studies established that the IMPDH‐encoding gene found in the MPA pathway confers resistance of MPA . This observation suggests that the compound produced by pyr cluster in P. thymicola may have similar IMPDH‐inhibiting bioactivities.…”

“…Interestingly, the P. thymicola genome encodes two copies of IMPDHs (a housekeeping copy and orf1 ). This is similar to the genomes of mycophenolic acid (MPA)‐producing strains . MPA is a known inhibitor of IMPDH, and previous studies established that the IMPDH‐encoding gene found in the MPA pathway confers resistance of MPA .…”

The Biosynthetic Gene Cluster for Andrastin A in Penicillium roqueforti

“…The strategy was essentially the same described by Del-Cid et al (2016). Briefly, a small sequence of each gene was amplified by PCR using suitable primers (Supplementary Table S1).…”

Compartmentalized Biosynthesis of Mycophenolic Acid

“…However, the genetic and enzymological bases for MPA biosynthesis remained obscure until the recent independent discoveries of three analogous MPA biosynthetic gene clusters ( SI Appendix , Fig. S2) (14–16). Upon identification of these clusters, a sub-set of the MPA biosynthetic pathway steps have been revealed through the functional characterization of three biosynthetic enzymes: MpaC (14, 17) and MpaDE (18) from P. brevicompactum IBT23078 as well as MpaG' from P. brevicompactum NRRL864 ( Pb 864 ) (15) (Fig.…”