Abstract
Filamentous fungi have long been recognized to be a rich source of secondary metabolites with potential medicinal applications. The recent genomic sequencing of several Aspergillus species has revealed that many secondary metabolite gene clusters are apparently silent under standard laboratory conditions. Several successful approaches have been utilized to upregulate these genes and unearth the corresponding natural products. A straightforward, reliable method to purify and characterize new metabolites therefore should be useful. Details are provided herein on the cultivation of Aspergillus nidulans and the LC/MS analysis of the metabolic profile. Following is an explanation of silica gel chromatography, HPLC, and preparative TLC. Finally, the NMR characterization of previously unknown A. nidulans metabolites is detailed.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Kennedy J, Auclair K, Kendrew SG, Park C, Vederas JC, Hutchinson CR (1999) Modulation of polyketide synthase activity by accessory proteins during lovastatin biosynthesis. Science 284:1368–1372
Payne GA, Nystrom GJ, Bhatnagar D, Cleveland TE, Woloshuk CP (1993) Cloning of the afl-2 gene involved in aflatoxin biosynthesis from Aspgergillus flavus. Appl Environ Microbiol 59:156–162
Galagan JE, Calvo SE, Cuomo C, Ma LJ, Wortman JR, Batzoglou S et al (2005) Sequencing of Aspergillus nidulans and comparative analysis with A-fumigatus and A-oryzae. Nature 438:1105–1115
Machida M, Asai K, Sano M, Tanaka T, Kumagai T, Terai G et al (2005) Genome sequencing and analysis of Aspergillus oryzae. Nature 438:1157–1161
Nierman WC, Pain A, Anderson MJ, Wortman JR, Kim HS, Arroyo J et al (2005) Genomic sequence of the pathogenic and allergenic filamentous fungus Aspergillus fumigatus. Nature 438:1151–1156
Pel HJ, de Winde JH, Archer DB, Dyer PS, Hofmann G, Schaap PJ et al (2007) Genome sequencing and analysis of the versatile cell Âfactory Aspergillus niger CBS 513.88. Nat Biotechnol 25:221–231
Bok JW, Chiang YM, Szewczyk E, Reyes-Dominguez Y, Davidson AD, Sanchez JF et al (2009) Chromatin-level regulation of biosynthetic gene clusters. Nat Chem Biol 5:462–464
Sanchez JF, Chiang YM, Szewczyk E, Davidson AD, Ahuja M, Oakley CE et al (2010) Molecular genetic analysis of the orsellinic acid/F9775 gene cluster of Aspergillus nidulans. Mol Biosyst 6:587–593
Szewczyk E, Chiang YM, Oakley CE, Davidson AD, Wang CCC, Oakley BR (2008) Identification and characterization of the asperthecin gene cluster of Aspergillus nidulans. Appl Environ Microbiol 74:7607–7612
Chiang YM, Szewczyk E, Davidson AD, Keller N, Oakley BR, Wang CCC (2009) A gene cluster containing two fungal polyketide synthases encodes the biosynthetic pathway for a polyketide, asperfuranone, in Aspergillus nidulans. J Am Chem Soc 131:2965–2970
Still WC, Kahn M, Mitra A (1978) Rapid chromatographic technique for preparative separations with moderate resolution. J Org Chem 43:2923–2925
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Sanchez, J.F., Wang, C.C.C. (2012). The Chemical Identification and Analysis of Aspergillus nidulans Secondary Metabolites. In: Keller, N., Turner, G. (eds) Fungal Secondary Metabolism. Methods in Molecular Biology, vol 944. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-122-6_6
Download citation
DOI: https://doi.org/10.1007/978-1-62703-122-6_6
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-121-9
Online ISBN: 978-1-62703-122-6
eBook Packages: Springer Protocols