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Inducer-independent production of pectinases in Aspergillus niger by overexpression of the D-galacturonic acid-responsive transcription factor gaaR
- Alazi, Ebru, Knetsch, Tim, Di Falco, Marcos, Reid, IanD., Arentshorst, Mark, Visser, Jaap, Tsang, Adrian, Ram, ArthurF. J.
- Applied microbiology and biotechnology 2018 v.102 no.6 pp. 2723-2736
- Aspergillus nidulans, Aspergillus niger, biochemical pathways, carbon, fructose, gene overexpression, genes, metabolism, pectins, polygalacturonase, transcription (genetics), transcription factors, transporters
- The transcription factor GaaR is needed for the expression of genes required for pectin degradation and transport and catabolism of the main degradation product, D-galacturonic acid (GA) in Aspergillus niger. In this study, we used the strong constitutive gpdA promoter of Aspergillus nidulans to overexpress gaaR in A. niger. Overexpression of gaaR resulted in an increased transcription of the genes encoding pectinases, (putative) GA transporters, and catabolic pathway enzymes even under non-inducing conditions, i.e., in the absence of GA. Exoproteome analysis of a strain overexpressing gaaR showed that this strain secretes highly elevated levels of pectinases when grown in fructose. The genes encoding exo-polygalacturonases were found to be subjected to CreA-mediated carbon catabolite repression, even in the presence of fructose. Deletion of creA in the strain overexpressing gaaR resulted in a further increase in pectinase production in fructose. We showed that GaaR localizes mainly in the nucleus regardless of the presence of an inducer, and that overexpression of gaaR leads to an increased concentration of GaaR in the nucleus.