Identification of enzymes and quantification of metabolic fluxes in the wild type and in a recombinant aspergillus oryzae strain

Applied and Environmental Microbiology
H PedersenJ Nielsen

Abstract

Two alpha-amylase-producing strains of Aspergillus oryzae, a wild-type strain and a recombinant containing additional copies of the alpha-amylase gene, were characterized with respect to enzyme activities, localization of enzymes to the mitochondria or cytosol, macromolecular composition, and metabolic fluxes through the central metabolism during glucose-limited chemostat cultivations. Citrate synthase and isocitrate dehydrogenase (NAD) activities were found only in the mitochondria, glucose-6-phosphate dehydrogenase and glutamate dehydrogenase (NADP) activities were found only in the cytosol, and isocitrate dehydrogenase (NADP), glutamate oxaloacetate transaminase, malate dehydrogenase, and glutamate dehydrogenase (NAD) activities were found in both the mitochondria and the cytosol. The measured biomass components and ash could account for 95% (wt/wt) of the biomass. The protein and RNA contents increased linearly with increasing specific growth rate, but the carbohydrate and chitin contents decreased. A metabolic model consisting of 69 fluxes and 59 intracellular metabolites was used to calculate the metabolic fluxes through the central metabolism at several specific growth rates, with ammonia or nitrate as the nitrogen sourc...Continue Reading

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Citations

Mar 31, 2011·Biotechnology and Bioengineering·Wanwipa VongsangnakJens Nielsen
Jan 15, 2014·Applied Microbiology and Biotechnology·Christoph KnufJens Nielsen
Aug 10, 1999·Biotechnology and Bioengineering·A L Santerre HenriksenJ Nielsen
Mar 8, 2000·Microbiology and Molecular Biology Reviews : MMBR·S OstergaardJ Nielsen
Jan 27, 2009·Applied and Environmental Microbiology·Gianni PanagiotouLisbeth Olsson
Nov 19, 2003·European Journal of Biochemistry·Helga DavidJens Nielsen
Jan 7, 2015·Journal of Industrial Microbiology & Biotechnology·Tobias KleinElmar Heinzle
Jul 31, 2013·Applied and Environmental Microbiology·Christoph KnufJens Nielsen
Jul 17, 2007·Bioprocess and Biosystems Engineering·R GheshlaghiP L Douglas
Jan 10, 2018·Microbial Biotechnology·Mariane Paludetti ZubietaAndré Ricardo de Lima Damasio
Aug 25, 2018·World Journal of Microbiology & Biotechnology·Sayak MitraBiswanath Mahanty

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