Proteomics

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QUANTITATIVE PROTEOME PROFILING OF NEUROSPORA CRASSA REVEALS A CELLOBIOSE-DEPENDENT NETWORK COORDINATING LIGNOCELLULOLYTIC RESPONSES


ABSTRACT: Cellulose, particularly the major cellulolytic product cellobiose, can induce the production of enzymes associated with deconstruction of lignocellulose in filamentous fungi. However, the detailed mechanisms underlying this biotechnologically important process remain to be disclosed. Here, the proteome response to cellobiose, crystalline cellulose (Avicel), and carbon starvation of a Neurospora crassa triple β-glucosidase mutant were compared using tandem mass tag (TMT)-based proteome quantification. Improved quantification accuracy was achieved with synchronous precursor selection (SPS)-based MS3 technology compared to MS2 using a high resolution tribrid mass spectrometer. Exposure to carbon starvation, cellobiose or Avicel induced the production of cellulase and lytic polysaccharide monooxygenase enzymes in N. crassa, as well as a cellobionic acid transporter, indicating their functional roles in the early adaptation to plant cell wall. In particular, cellobiose specifically induced the production of proteins in the functional categories of protein processing and export as well as cell wall organization. The data presented here integrates the signaling pathway associated with cellobiose transporters CDT-1 and/or CDT-2 with the direct targets of the transcription factors CLR-1, CLR-2, and XLR-1, the unfolded protein response (UPR) mediated by Ire-1/Hac-1, as well as calcium homeostasis and cell wall organization. The cellobiose-dependent response network will be useful for rational strain improvement to facilitate the production of lignocellulases in filamentous fungi and plant biomass-based products.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Neurospora Crassa Or74a

SUBMITTER: Shaolin Chen  

LAB HEAD: Shaolin Chen

PROVIDER: PXD011598 | Pride | 2020-05-15

REPOSITORIES: Pride

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Publications

Quantitative Proteome Profiling Reveals Cellobiose-Dependent Protein Processing and Export Pathways for the Lignocellulolytic Response in Neurospora crassa.

Liu Dan D   Liu Yisong Y   Zhang Duoduo D   Chen Xiaoting X   Liu Qian Q   Xiong Bentao B   Zhang Lihui L   Wei Linfang L   Wang Yifan Y   Fang Hao H   Liesche Johannes J   Wei Yahong Y   Glass N Louise NL   Hao Zhiqi Z   Chen Shaolin S  

Applied and environmental microbiology 20200720 15


Filamentous fungi are intensively used for producing industrial enzymes, including lignocellulases. Employing insoluble cellulose to induce the production of lignocellulases causes some drawbacks, e.g., a complex fermentation operation, which can be overcome by using soluble inducers such as cellobiose. Here, a triple β-glucosidase mutant of <i>Neurospora crassa</i>, which prevents rapid turnover of cellobiose and thus allows the disaccharide to induce lignocellulases, was applied to profile the  ...[more]

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