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Title: Identification of a classical mutant in the industrial host Aspergillus niger by systems genetics: LaeA is required for citric acid production and regulates the formation of some secondary metabolites

Abstract

The asexual filamentous fungus Aspergillus niger is an important industrial cell factory for citric acid production. In this study, we genetically characterized a UV-generated A. niger mutant that was originally isolated as a nonacidifying mutant, which is a desirable trait for industrial enzyme production. Physiological analysis showed that this mutant did not secrete large amounts of citric acid and oxalic acid, thus explaining the nonacidifying phenotype. As traditional complementation approaches to characterize the mutant genotype were unsuccessful, we used bulk segregant analysis in combination with high-throughput genome sequencing to identify the mutation responsible for the nonacidifying phenotype. Since A. niger has no sexual cycle, parasexual genetics was used to generate haploid segregants derived from diploids by loss of whole chromosomes. We found that the nonacidifying phenotype was caused by a point mutation in the laeA gene. LaeA encodes a putative methyltransferase-domain protein, which we show here to be required for citric acid production in an A. niger lab strain (N402) and in other citric acid production strains. The unexpected link between LaeA and citric acid production could provide new insights into the transcriptional control mechanisms related to citric acid production in A. niger. Interestingly, the secondary metabolite profile of amore » ΔlaeA strain differed from the wild-type strain, showing both decreased and increased metabolite levels, indicating that LaeA is also involved in regulating the production of secondary metabolites. As a result, we show that our systems genetics approach is a powerful tool to identify trait mutations.« less

Authors:
 [1];  [1];  [1];  [2];  [2];  [3];  [3];  [4];  [1]
  1. Leiden Univ., Leiden (The Netherlands)
  2. Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
  3. Technical Univ. of Denmark, Lyngby (Denmark)
  4. Leiden Univ., Leiden (The Netherlands); Dutch DNA Biotech (The Netherlands)
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1249369
Report Number(s):
PNNL-SA-114553
Journal ID: ISSN 2160-1836
Grant/Contract Number:  
AC05-76RL01830
Resource Type:
Accepted Manuscript
Journal Name:
G3
Additional Journal Information:
Journal Volume: 6; Journal Issue: 1; Journal ID: ISSN 2160-1836
Publisher:
Genetics Society of America
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; organic acids; filamentous fungi; bulk segregant analysis; parasexual cycle; genome sequencing

Citation Formats

Niu, Jing, Arentshorst, Mark, Nair, P. Deepa S., Dai, Ziyu, Baker, Scott E., Frisvad, Jens C., Nielsen, Kristian F., Punt, Peter J., and Ram, Arthur F. J. Identification of a classical mutant in the industrial host Aspergillus niger by systems genetics: LaeA is required for citric acid production and regulates the formation of some secondary metabolites. United States: N. p., 2015. Web. doi:10.1534/g3.115.024067.
Niu, Jing, Arentshorst, Mark, Nair, P. Deepa S., Dai, Ziyu, Baker, Scott E., Frisvad, Jens C., Nielsen, Kristian F., Punt, Peter J., & Ram, Arthur F. J. Identification of a classical mutant in the industrial host Aspergillus niger by systems genetics: LaeA is required for citric acid production and regulates the formation of some secondary metabolites. United States. doi:10.1534/g3.115.024067.
Niu, Jing, Arentshorst, Mark, Nair, P. Deepa S., Dai, Ziyu, Baker, Scott E., Frisvad, Jens C., Nielsen, Kristian F., Punt, Peter J., and Ram, Arthur F. J. Fri . "Identification of a classical mutant in the industrial host Aspergillus niger by systems genetics: LaeA is required for citric acid production and regulates the formation of some secondary metabolites". United States. doi:10.1534/g3.115.024067. https://www.osti.gov/servlets/purl/1249369.
@article{osti_1249369,
title = {Identification of a classical mutant in the industrial host Aspergillus niger by systems genetics: LaeA is required for citric acid production and regulates the formation of some secondary metabolites},
author = {Niu, Jing and Arentshorst, Mark and Nair, P. Deepa S. and Dai, Ziyu and Baker, Scott E. and Frisvad, Jens C. and Nielsen, Kristian F. and Punt, Peter J. and Ram, Arthur F. J.},
abstractNote = {The asexual filamentous fungus Aspergillus niger is an important industrial cell factory for citric acid production. In this study, we genetically characterized a UV-generated A. niger mutant that was originally isolated as a nonacidifying mutant, which is a desirable trait for industrial enzyme production. Physiological analysis showed that this mutant did not secrete large amounts of citric acid and oxalic acid, thus explaining the nonacidifying phenotype. As traditional complementation approaches to characterize the mutant genotype were unsuccessful, we used bulk segregant analysis in combination with high-throughput genome sequencing to identify the mutation responsible for the nonacidifying phenotype. Since A. niger has no sexual cycle, parasexual genetics was used to generate haploid segregants derived from diploids by loss of whole chromosomes. We found that the nonacidifying phenotype was caused by a point mutation in the laeA gene. LaeA encodes a putative methyltransferase-domain protein, which we show here to be required for citric acid production in an A. niger lab strain (N402) and in other citric acid production strains. The unexpected link between LaeA and citric acid production could provide new insights into the transcriptional control mechanisms related to citric acid production in A. niger. Interestingly, the secondary metabolite profile of a ΔlaeA strain differed from the wild-type strain, showing both decreased and increased metabolite levels, indicating that LaeA is also involved in regulating the production of secondary metabolites. As a result, we show that our systems genetics approach is a powerful tool to identify trait mutations.},
doi = {10.1534/g3.115.024067},
journal = {G3},
number = 1,
volume = 6,
place = {United States},
year = {2015},
month = {11}
}

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