Abstract
Aspergillus niger is used by the industry to produce enzymes and metabolites such as citric acid. In liquid cultures, it can grow as a dispersed mycelium or as micro-colonies with a width in the micrometer to millimeter range. Here, it was assessed whether expression of genes encoding secreted enzymes depends on mycelium morphology. To this end, expression of the reporter gene gfp from the promoters of the glucoamylase gene glaA, the feruloyl esterase gene faeA and the α-glucuronidase gene aguA was causally related to micro-colony size within a liquid shaken culture. Data could be fitted by hyperbolic functions, implying that the genes encoding these secreted proteins are expressed in a shell at the periphery of the micro-colony. The presence of such a shell was confirmed by confocal microscopy. Modelling predicted that the width of these zones was 13 to 156 µm depending on growth medium and micro-colony diameter. Together, data indicate that the highest productive micro-colonies are those colonies that have a radius ≤ the width of the peripheral expression zone.
Highlights
Aspergillus niger is used by the industry to produce enzymes and metabolites such as citric acid
Wild-type strain N402 was grown in CMX for 16 h followed by a transfer to either CMX, CMM, mM xylose (MMX) or MMM
Morphology of the mycelium in liquid cultures such as bioreactors depends on various factors including inoculum size, surface composition of the inoculum, medium composition, and mixing conditions[1,7,11]
Summary
Aspergillus niger is used by the industry to produce enzymes and metabolites such as citric acid In liquid cultures, it can grow as a dispersed mycelium or as micro-colonies with a width in the micrometer to millimeter range. It can grow as a dispersed mycelium or as micro-colonies with a width in the micrometer to millimeter range It was assessed whether expression of genes encoding secreted enzymes depends on mycelium morphology. We here studied whether micro-colony diameter affects expression of genes encoding secreted proteins in A. niger To this end, the highly expressed genes encoding feruloyl esterase FaeA7,8, α-glucuronidase AguA7,9 and glucoamylase GlaA7,10 were used as model genes of xylanolytic (FaeA, AguA) and amylolytic (GlaA) activity. Results imply that small micro-colonies of A. niger produce more secreted proteins per biomass than large micro-colonies
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