title: An efficient system for heterologous expression of secondary metabolite genes in Aspergillus nidulans.
abstract: Fungal secondary metabolites (SMs) are an important source of medically valuable compounds. Genome projects have revealed that fungi have many SM biosynthetic gene clusters that are not normally expressed. To access these potentially valuable, cryptic clusters, we have developed a heterologous expression system in Aspergillus nidulans . We have developed an efficient system for amplifying genes from a target fungus, placing them under control of a regulatable promoter, transferring them into A. nidulans , and expressing them. We have validated this system by expressing nonreducing polyketide synthases of Aspergillus terreus and additional genes required for compound production and release. We have obtained compound production and release from six of these nonreducing polyketide synthases and have identified the products. To demonstrate that the procedure allows transfer and expression of entire secondary metabolite biosynthetic pathways, we have expressed all the genes of a silent A. terreus cluster and demonstrate that it produces asperfuranone. Further, by expressing the genes of this pathway in various combinations, we have clarified the asperfuranone biosynthetic pathway. We have also developed procedures for deleting entire A. nidulans SM clusters. This allows us to remove clusters that might interfere with analyses of heterologously expressed genes and to eliminate unwanted toxins.
authors.affiliation: Department of Pharmacology and Pharmaceutical Sciences, School of Pharmacy, University of Southern California, 1985 Zonal Avenue, Los Angeles, California 90089, USA.
authors.forename: Yi-Ming
authors.initials: YM
authors.lastname: Chiang
authors.forename: C Elizabeth
authors.initials: CE
authors.lastname: Oakley
authors.forename: Manmeet
authors.initials: M
authors.lastname: Ahuja
authors.forename: Ruth
authors.initials: R
authors.lastname: Entwistle
authors.forename: Aric
authors.initials: A
authors.lastname: Schultz
authors.forename: Shu-Lin
authors.initials: SL
authors.lastname: Chang
authors.forename: Calvin T
authors.initials: CT
authors.lastname: Sung
authors.forename: Clay C C
authors.initials: CC
authors.lastname: Wang
authors.forename: Berl R
authors.initials: BR
authors.lastname: Oakley
mesh_terms: D001233:Aspergillus nidulans / Q000235:genetics* / Q000378:metabolism; D001572:Benzofurans / Q000737:chemistry / Q000378:metabolism*; D008968:Molecular Conformation; D048630:Polyketide Synthases / Q000378:metabolism
chemical_list: D001572:Benzofurans; C552682:asperfuranone; D048630:Polyketide Synthases