Unveiling a classical mutant in the context of the GH3 β-glucosidase family in Neurospora crassa
Abstract
Classical fungal mutant strains obtained by mutagenesis have helped to elucidate fundamental metabolic pathways in the past. In the filamentous fungus Neurospora crassa, the gluc-1 strain was isolated long ago and characterized by its low level of β-glucosidase activity, which is essential for the degradation of cellulose, the most abundant biopolymer on Earth and the main polymeric component of the plant cell wall. Based on genomic resequencing, we hypothesized that the causative mutation resides in the β-glucosidase gene gh3-3 (bgl6, NCU08755). In this work, growth patterns, enzymatic activities and sugar utilization rates were analyzed in several mutant and overexpression strains related to gluc-1 and gh3-3. In addition, different mutants affected in the degradation and transport of cellobiose were analyzed. While overexpression of gh3-3 led to the recovery of β-glucosidase activity in the gluc-1 mutant, as well as normal utilization of cellobiose, the full gene deletion strain Δgh3-3 was found to behave differently than gluc-1 with lower secreted β-glucosidase activity, indicating a dominant role of the amino acid substitution in the point mutated gh3-3 gene of gluc-1. Our results furthermore confirm that GH3-3 is the major extracellular β-glucosidase in N. crassa and demonstrate that the two cellodextrin transporters CDT-1 and CDT-2more »
- Authors:
-
[1]
- Technical University of Munich (Germany)
- Joint BioEnergy Institute (JBEI), Emeryville, CA (United States); Pacific Northwest National Laboratory (PNNL), Richland, WA (United States)
- Pacific Northwest National Laboratory (PNNL), Richland, WA (United States)
- Chinese Academy of Sciences (CAS), Tianjin (China); National Technology Innovation Center of Synthetic Biology, Tianjin (China)
- Publication Date:
- Research Org.:
- Pacific Northwest National Laboratory (PNNL), Richland, WA (United States)
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 2282268
- Report Number(s):
- PNNL-SA-194079
Journal ID: ISSN 2191-0855
- Grant/Contract Number:
- AC05-76RL01830
- Resource Type:
- Accepted Manuscript
- Journal Name:
- AMB Express
- Additional Journal Information:
- Journal Volume: 14; Journal Issue: 1; Journal ID: ISSN 2191-0855
- Publisher:
- Springer
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 60 APPLIED LIFE SCIENCES; β-glucosidase; gluc-1 mutant; Neurospora crassa; Cellobiose utilization
Citation Formats
Zhang, Yuxin, Nada, Basant, Baker, Scott E., Evans, James E., Tian, Chaoguang, Benz, J. Philipp, and Tamayo, Elisabeth. Unveiling a classical mutant in the context of the GH3 β-glucosidase family in Neurospora crassa. United States: N. p., 2024.
Web. doi:10.1186/s13568-023-01658-0.
Zhang, Yuxin, Nada, Basant, Baker, Scott E., Evans, James E., Tian, Chaoguang, Benz, J. Philipp, & Tamayo, Elisabeth. Unveiling a classical mutant in the context of the GH3 β-glucosidase family in Neurospora crassa. United States. https://doi.org/10.1186/s13568-023-01658-0
Zhang, Yuxin, Nada, Basant, Baker, Scott E., Evans, James E., Tian, Chaoguang, Benz, J. Philipp, and Tamayo, Elisabeth. Fri .
"Unveiling a classical mutant in the context of the GH3 β-glucosidase family in Neurospora crassa". United States. https://doi.org/10.1186/s13568-023-01658-0. https://www.osti.gov/servlets/purl/2282268.
@article{osti_2282268,
title = {Unveiling a classical mutant in the context of the GH3 β-glucosidase family in Neurospora crassa},
author = {Zhang, Yuxin and Nada, Basant and Baker, Scott E. and Evans, James E. and Tian, Chaoguang and Benz, J. Philipp and Tamayo, Elisabeth},
abstractNote = {Classical fungal mutant strains obtained by mutagenesis have helped to elucidate fundamental metabolic pathways in the past. In the filamentous fungus Neurospora crassa, the gluc-1 strain was isolated long ago and characterized by its low level of β-glucosidase activity, which is essential for the degradation of cellulose, the most abundant biopolymer on Earth and the main polymeric component of the plant cell wall. Based on genomic resequencing, we hypothesized that the causative mutation resides in the β-glucosidase gene gh3-3 (bgl6, NCU08755). In this work, growth patterns, enzymatic activities and sugar utilization rates were analyzed in several mutant and overexpression strains related to gluc-1 and gh3-3. In addition, different mutants affected in the degradation and transport of cellobiose were analyzed. While overexpression of gh3-3 led to the recovery of β-glucosidase activity in the gluc-1 mutant, as well as normal utilization of cellobiose, the full gene deletion strain Δgh3-3 was found to behave differently than gluc-1 with lower secreted β-glucosidase activity, indicating a dominant role of the amino acid substitution in the point mutated gh3-3 gene of gluc-1. Our results furthermore confirm that GH3-3 is the major extracellular β-glucosidase in N. crassa and demonstrate that the two cellodextrin transporters CDT-1 and CDT-2 are essential for growth on cellobiose when the three main N. crassa β-glucosidases are absent. Overall, these findings provide valuable insight into the mechanisms of cellulose utilization in filamentous fungi, being an essential step in the efficient production of biorefinable sugars from agricultural and forestry plant biomass.},
doi = {10.1186/s13568-023-01658-0},
journal = {AMB Express},
number = 1,
volume = 14,
place = {United States},
year = {Fri Jan 05 00:00:00 EST 2024},
month = {Fri Jan 05 00:00:00 EST 2024}
}
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