Rational elimination of Aspergillus terreus sulochrin production.
Elimination of undesirable co-metabolites from industrial fermentations is often required due to the toxicities associated with the contaminants and/or due to difficulties in removing the contaminants during downstream processing. Sulochrin is a co-metabolite produced during the Aspergillus terreus lovastatin fermentation. Examination of the sulochrin biosynthetic pathway identifies the emodin anthrone polyketide synthase (PKS) at the origin. Thus, genetically disrupting the emodin anthrone PKS gene was expected to result in the elimination of sulochrin biosynthesis. To perform the disruption by homologous recombination, a fragment of the emodin anthrone PKS gene first needed to be isolated. Analysis of several reported fungal PKS amino acid sequences has identified three subfamilies of related sequences (called the Patulin subfamily, the Pigment subfamily, and the Reduction subfamily). PCR primers specific for the Pigment subfamily (of which the emodin anthrone PKS is expected to belong) were used to isolate a fragment of a novel PKS gene from A. terreus. Targeted gene disruption identifies the novel gene fragment as that from the emodin anthrone PKS. Consequently, the gene disruption event eliminated the production of metabolites from the sulochrin biosynthetic pathway.
Couch
Robin D
RD
Division of Biochemistry, Department of Biological Sciences, University of Calgary, Calgary, Alberta, Canada T2N 1N4. Robin.D.Couch@dartmouth.edu
Gaucher
G Maurice
GM
D001230:Aspergillus / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D001565:Benzoates / Q000378:metabolism*; D004642:Emodin / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal / Q000235:genetics; D020868:Gene Silencing; D024861:Genetic Enhancement / Q000379:methods*; D008148:Lovastatin / Q000096:biosynthesis*; D009097:Multienzyme Complexes / Q000235:genetics* / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000378:metabolism
D001565:Benzoates; D009097:Multienzyme Complexes; D011994:Recombinant Proteins; C111298:sulochrin; D008148:Lovastatin; D004642:Emodin