Inducer-independent production of pectinases in Aspergillus niger by overexpression of the D-galacturonic acid-responsive transcription factor gaaR

被引:29
作者
Alazi, Ebru [1 ]
Knetsch, Tim [1 ]
Di Falco, Marcos [2 ]
Reid, Ian D. [2 ]
Arentshorst, Mark [1 ]
Visser, Jaap [1 ]
Tsang, Adrian [2 ]
Ram, Arthur F. J. [1 ]
机构
[1] Leiden Univ, Inst Biol Leiden, Mol Microbiol & Biotechnol, Sylviusweg 72, NL-2333 BE Leiden, Netherlands
[2] Concordia Univ, Ctr Struct & Funct Genom, Quebec City, PQ, Canada
基金
加拿大自然科学与工程研究理事会;
关键词
Transcriptome; Exoproteome; Gene regulation; Transcription factor localization; GFP fluorescence; Transcription factor concentration; TRICHODERMA-REESEI; GENE-CLUSTER; ACTIVATOR; INDUCTION; NIDULANS; STRAINS; REPRESSION; REGULATOR; ENZYMES; MARKER;
D O I
10.1007/s00253-018-8753-7
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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.
引用
收藏
页码:2723 / 2736
页数:14
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