Transcriptional analysis of genes for energy catabolism and hydrolytic enzymes in the filamentous fungus Aspergillus oryzae using cDNA microarrays and expressed sequence tags

Aspergillus oryzae is a fungus used extensively in the fermentation industry. We constructed cDNA microarrays comprising 2,070 highly expressed cDNAs selected from the ∼6,000 non-redundant expressed sequence tags (ESTs) in the A. oryzae EST database (http://www.aist.go.jp/RIODB/ffdb/index.html). Using the cDNA microarrays, we analyzed the gene expression profiles of A. oryzae cells grown under the glucose-rich (AC) and glucose-depleted (AN) liquid culture conditions used during the construction of the EST database. The sets of genes identified by the cDNA microarray as highly expressed under each culture condition agreed well with the highly redundant ESTs obtained under the same conditions. In particular, transcription levels of most catabolic genes of the glycolytic pathway (EMP) and tricarboxylic acid (TCA) cycle were higher under AC than AN conditions, suggesting that A. oryzae uses both EMP and TCA for glucose metabolism under AC conditions. We further studied the expression of genes encoding hydrolytic enzymes and enzymes involved in energy catabolism by using three industrial solid-phase biomass media, including wheat-bran. The wheat-bran culture gave the richest gene expression profile of hydrolytic enzymes and the lowest expression levels of catabolic genes (EMP, TCA) among the three media tested. The low expression levels of catabolic genes in the wheat-bran culture may release catabolite repression, consequently leading to the rich expression profiles of the hydrolytic enzymes.

[1]  R. Ory,et al.  Hemicellulose composition of dietary fiber of milled rice and rice bran. , 1978, Journal of agricultural and food chemistry.

[2]  M. Taylor,et al.  Applications of microbial enzymes in food systems and in biotechnology. , 1979, Advances in applied microbiology.

[3]  Hisaharu Taguchi,et al.  Growth and Enzyme Production in a Solid-State Culture of Aspergillus oryzae : , 1982 .

[4]  J. Martial,et al.  A method for isolation of intact, translationally active ribonucleic acid. , 1983, DNA.

[5]  L. Thim,et al.  High Level Expression of Recombinant Genes in Aspergillus Oryzae , 1988, Bio/Technology.

[6]  J. Kelly,et al.  Specific binding sites in the alcR and alcA promoters of the ethanol regulon for the CREA repressor mediating carbon cataboiite repression in Aspergillus nidulans , 1993, Molecular microbiology.

[7]  D. Mackenzie,et al.  A truncated glucoamylase gene fusion for heterologous protein secretion from Aspergillus niger. , 1993, FEMS microbiology letters.

[8]  C. Scazzocchio,et al.  Two different, adjacent and divergent zinc finger binding sites are necessary for CREA‐mediated carbon catabolite repression in the proline gene cluster of Aspergillus nidulans. , 1994, The EMBO journal.

[9]  M. Machida,et al.  Molecular cloning of a cDNA encoding enolase from the filamentous fungus, Aspergillus oryzae , 1996, Current Genetics.

[10]  P. Brown,et al.  Exploring the metabolic and genetic control of gene expression on a genomic scale. , 1997, Science.

[11]  P. Punt,et al.  Efficient production of secreted proteins by Aspergillus : progress, limitations and prospects , 1997, Applied Microbiology and Biotechnology.

[12]  J. Gancedo Yeast Carbon Catabolite Repression , 1998, Microbiology and Molecular Biology Reviews.

[13]  R. Kanamoto,et al.  Soybean curd refuse alleviates experimental tumorigenesis in rat colon. , 1999, Bioscience, biotechnology, and biochemistry.

[14]  E. Nakano,et al.  Cloning and expression of a cDNA encoding the laccase from Schizophyllum commune. , 1999, Bioscience, biotechnology, and biochemistry.

[15]  J. Latgé,et al.  Aspergillus fumigatus and Aspergillosis , 1999, Clinical Microbiology Reviews.

[16]  J. Pronk,et al.  Regulation of pyruvate metabolism in chemostat cultures of Kluyveromyces lactis CBS 2359 , 2000, Yeast.

[17]  Tomoo Suzuki,et al.  Comprehensive cloning and expression analysis of glycolytic genes from the filamentous fungus, Aspergillus oryzae , 2000, Current Genetics.

[18]  Antonio Carlos Augusto da Costa,et al.  Hydrolytic enzyme production in solid-state fermentation by Aspergillus niger 3T5B8 , 2000 .

[19]  T. Petit,et al.  Carbohydrate and energy-yielding metabolism in non-conventional yeasts. , 2000, FEMS microbiology reviews.

[20]  J. Bennett,et al.  Genomics for Fungi , 2001 .

[21]  M. Flipphi,et al.  Regulation of the Aldehyde Dehydrogenase Gene (aldA) and Its Role in the Control of the Coinducer Level Necessary for Induction of the Ethanol Utilization Pathway in Aspergillus nidulans * , 2001, The Journal of Biological Chemistry.

[22]  H. Sealy-Lewis,et al.  ADHII in Aspergillus nidulans is induced by carbon starvation stress. , 2001, Fungal genetics and biology : FG & B.

[23]  K. Abe,et al.  chsZ, a gene for a novel class of chitin synthase from Aspergillus oryzae , 2002, Current Genetics.

[24]  Jiujiang Yu,et al.  Nonfunctionality of Aspergillus sojae aflR in a Strain of Aspergillus parasiticus with a Disrupted aflR Gene , 2002, Applied and Environmental Microbiology.

[25]  M. Machida Progress of Aspergillus oryzae genomics. , 2002, Advances in applied microbiology.

[26]  O. Akita,et al.  Subtractive cloning of cDNA from Aspergillus oryzae differentially regulated between solid-state culture and liquid (submerged) culture , 2002, Current Genetics.

[27]  Ari J. S. Ferreira,et al.  Elucidation of the Metabolic Fate of Glucose in the Filamentous Fungus Trichoderma reesei Using Expressed Sequence Tag (EST) Analysis and cDNA Microarrays* , 2002, The Journal of Biological Chemistry.

[28]  J. Visser,et al.  Onset of Carbon Catabolite Repression in Aspergillus nidulans , 2003, The Journal of Biological Chemistry.

[29]  K. Kitamoto,et al.  High level expression of the synthetic human lysozyme gene in Aspergillus oryzae , 1992, Applied Microbiology and Biotechnology.

[30]  W. Nierman,et al.  10 - Genomics of Economically Significant Aspergillus and Fusarium Species , 2004 .

[31]  K. Kitamoto,et al.  Secretion of calf chymosin from the filamentous fungus Aspergillus oryzae , 1993, Applied Microbiology and Biotechnology.

[32]  Hans Peter Heldt-Hansen,et al.  On the safety of Aspergillus oryzae: a review , 1992, Applied Microbiology and Biotechnology.

[33]  J. Kelly,et al.  Alcohol dehydrogenase III inAspergillus nidulans is anaerobically induced and post-transcriptionally regulated , 1990, Molecular and General Genetics MGG.