Identification of Aspergillus niger Aquaporins Involved in Hydrogen Peroxide Signaling

Abstract

Aspergillus niger is a robust microbial cell factory for organic acid production. However, the regulation of many industrially important pathways is still poorly understood. The regulation of the glucose oxidase (Gox) expression system, involved in the biosynthesis of gluconic acid, has recently been uncovered. The results of that study show hydrogen peroxide, a by-product of the extracellular conversion of glucose to gluconate, has a pivotal role as a signaling molecule in the induction of this system. In this study, the facilitated diffusion of hydrogen peroxide via aquaporin water channels (AQPs) was studied. AQPs are transmembrane proteins of the major intrinsic proteins (MIPs) superfamily. In addition to water and glycerol, they may also transport small solutes such as hydrogen peroxide. The genome sequence of A. niger N402 was screened for putative AQPs. Seven AQPs were found and could be classified into three main groups. One protein (AQPA) belonged to orthodox AQP, three (AQPB, AQPD, and AQPE) were grouped in aquaglyceroporins (AQGP), two (AQPC and AQPF) were in X-intrinsic proteins (XIPs), and the other (AQPG) could not be classified. Their ability to facilitate diffusion of hydrogen peroxide was identified using yeast phenotypic growth assays and by studying AQP gene knock-outs in A. niger. The X-intrinsic protein AQPF appears to play roles in facilitating hydrogen peroxide transport across the cellular membrane in both Saccharomyces cerevisiae and A. niger experiments.

The Transcriptome Response to Azole Compounds in Aspergillus fumigatus Shows Differential Gene Expression across Pathways Essential for Azole Resistance and Cell Survival

“…In this study, the observed results were not validated by qPCR, as previous experiments by us and other groups have shown the validity of RNA-seq methods for Aspergillus spp. ([ 29 , 36 , 75 , 76 , 77 , 78 , 79 , 80 , 81 ]). In addition, the benchmark study conducted by Evereart et al, 2017 (Sci Rep) [ 82 ] showed a high concordance (~80%) between RNA-seq and qPCR data for various workflows and mapping tools.…”

Osmotically Activated Anion Current of Phycomyces Blakesleeanus—Filamentous Fungi Counterpart to Vertebrate Volume Regulated Anion Current

“…Aquaglyceroporins, which permeate small uncharged solutes and consequentially do not mediate ion currents, are expressed in the membrane of filamentous fungi [ 35 ]. Recently, seven homologues of the aquaglyceroporins have been identified in Aspergillus [ 36 ], but transport of small solutes has not been measured, except for the channel involvement in the hydrogen peroxide efflux.…”