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Different control mechanisms regulate glucoamylase and protease gene transcription in Aspergillus oryzae in solid-state and submerged fermentation

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Author: Biesebeke, R. te · Biezen, N. van · Vos, W.M. de · Hondel, C.A.M.J.J. van den · Punt, P.J.
Type:article
Date:2005
Institution: TNO Kwaliteit van Leven
Source:Applied Microbiology and Biotechnology, 1, 67, 75-82
Identifier: 238415
doi: doi:10.1007/s00253-004-1807-z
Keywords: Biology · Biotechnology · Carbon · Control theory · Crops · Enzymes · Fermentation · Genes · Nitrogen · Gene expressions · Gene transcription · Glucomylase (glaB) · Solid-state fermentation (SSF) · Enzyme kinetics · carbon · glucan 1,4 alpha glucosidase · nitrogen · proteinase · fermentation · gene expression · protein · article · Aspergillus oryzae · controlled study · culture medium · fungus hyphae · gene control · gene expression · liquid culture · molecular mechanics · nonhuman · pH measurement · seed kernel · solid state fermentation · transcription regulation · wheat · Aspergillus oryzae · Biomass · Carbon · Fermentation · Gene Expression Profiling · Gene Expression Regulation, Fungal · Glucan 1,4-alpha-Glucosidase · Glucose · Hydrogen-Ion Concentration · Nitrogen · Peptide Hydrolases · RNA, Fungal · RNA, Messenger · Transcription, Genetic · Triticum · Carbon · Enzymatic Activity · Enzymes · Fermentation · Genes · Nitrogen · Wheat · Aspergillus oryzae · Triticum aestivum

Abstract

Solid-state fermentation (SSF) with Aspergillus oryzae results in high levels of secreted protein. However, control mechanisms of gene expression in SSF have been only poorly studied. In this study we show that both glucoamylase (glaB) and protease (alpA, nptB) genes are highly expressed during surface cultivation on wheat-based solid medium, and even higher during cultivation on wheat kernels. In wheat-based liquid medium, low levels of gene expression are observed. Typical SSF cultivation conditions, such as low water activity and the formation of aerial hyphae, did not contribute to the high-level gene expression on wheat-based solid medium. Analysis of wheat-based solid and liquid cultivations showed differences in carbon and nitrogen utilisation and external pH. The results presented show that the difference in regulation of transcription of the alpA and nptB genes in wheat-based liquid and solid medium could be pH dependent, involving a pH-dependent transcription regulator. The results obtained suggest that the difference in regulation of transcription of the glaB gene in wheat-based liquid and solid medium is caused by a difference in carbohydrate degradation and consumption under the different culture conditions. © Springer-Verlag 2004.