Volatile profiling reveals intracellular metabolic changes in Aspergillus parasiticus: veA regulates branched chain amino acid and ethanol metabolism.
BACKGROUND
Filamentous fungi in the genus Aspergillus produce a variety of natural products, including aflatoxin, the most potent naturally occurring carcinogen known. Aflatoxin biosynthesis, one of the most highly characterized secondary metabolic pathways, offers a model system to study secondary metabolism in eukaryotes. To control or customize biosynthesis of natural products we must understand how secondary metabolism integrates into the overall cellular metabolic network. By applying a metabolomics approach we analyzed volatile compounds synthesized by Aspergillus parasiticus in an attempt to define the association of secondary metabolism with other metabolic and cellular processes.


RESULTS
Volatile compounds were examined using solid phase microextraction--gas chromatography/mass spectrometry. In the wild type strain Aspergillus parasiticus SU-1, the largest group of volatiles included compounds derived from catabolism of branched chain amino acids (leucine, isoleucine, and valine); we also identified alcohols, esters, aldehydes, and lipid-derived volatiles. The number and quantity of the volatiles produced depended on media composition, time of incubation, and light-dark status. A block in aflatoxin biosynthesis or disruption of the global regulator veA affected the volatile profile. In addition to its multiple functions in secondary metabolism and development, VeA negatively regulated catabolism of branched chain amino acids and synthesis of ethanol at the transcriptional level thus playing a role in controlling carbon flow within the cell. Finally, we demonstrated that volatiles generated by a veA disruption mutant are part of the complex regulatory machinery that mediates the effects of VeA on asexual conidiation and sclerotia formation.


CONCLUSIONS
1) Volatile profiling provides a rapid, effective, and powerful approach to identify changes in intracellular metabolic networks in filamentous fungi. 2) VeA coordinates the biosynthesis of secondary metabolites with catabolism of branched chain amino acids, alcohol biosynthesis, and β-oxidation of fatty acids. 3) Intracellular chemical development in A. parasiticus is linked to morphological development. 4) Understanding carbon flow through secondary metabolic pathways and catabolism of branched chain amino acids is essential for controlling and customizing production of natural products.
Roze
Ludmila V
LV
Department of Food Science and Human Nutrition, Michigan State University, East Lansing, MI, USA. roze@msu.edu
Chanda
Anindya
A
Laivenieks
Maris
M
Beaudry
Randolph M
RM
Artymovich
Katherine A
KA
Koptina
Anna V
AV
Awad
Deena W
DW
Valeeva
Dina
D
Jones
Arthur D
AD
Linz
John E
JE
D000348:Aflatoxins / Q000378:metabolism; D000597:Amino Acids, Branched-Chain / Q000378:metabolism*; D001230:Aspergillus / Q000378:metabolism*; D000431:Ethanol / Q000378:metabolism*; D005227:Fatty Acids / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D015227:Lipid Peroxidation; D014158:Transcription, Genetic; D055549:Volatile Organic Compounds / Q000737:chemistry*
D000348:Aflatoxins; D000597:Amino Acids, Branched-Chain; D005227:Fatty Acids; D005656:Fungal Proteins; D055549:Volatile Organic Compounds; D000431:Ethanol