Nitrogen limitation causes a seismic shift in redox state and phosphorylation of proteins implicated in carbon flux and lipidome remodeling in Rhodotorula toruloides

Abstract Background Oleaginous yeast are prodigious producers of oleochemicals, offering alternative and secure sources for applications in foodstuff, skincare, biofuels, and bioplastics. Nitrogen starvation is the primary strategy used to induce oil accumulation in oleaginous yeast as part of a glo...

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Main Authors: Austin Gluth, Jeffrey J. Czajka, Xiaolu Li, Kent J. Bloodsworth, Josie G. Eder, Jennifer E. Kyle, Rosalie K. Chu, Bin Yang, Wei-Jun Qian, Pavlo Bohutskyi, Tong Zhang
Format: Article
Language:English
Published: BMC 2025-07-01
Series:Biotechnology for Biofuels and Bioproducts
Online Access:https://doi.org/10.1186/s13068-025-02657-y
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author Austin Gluth
Jeffrey J. Czajka
Xiaolu Li
Kent J. Bloodsworth
Josie G. Eder
Jennifer E. Kyle
Rosalie K. Chu
Bin Yang
Wei-Jun Qian
Pavlo Bohutskyi
Tong Zhang
author_facet Austin Gluth
Jeffrey J. Czajka
Xiaolu Li
Kent J. Bloodsworth
Josie G. Eder
Jennifer E. Kyle
Rosalie K. Chu
Bin Yang
Wei-Jun Qian
Pavlo Bohutskyi
Tong Zhang
author_sort Austin Gluth
collection DOAJ
description Abstract Background Oleaginous yeast are prodigious producers of oleochemicals, offering alternative and secure sources for applications in foodstuff, skincare, biofuels, and bioplastics. Nitrogen starvation is the primary strategy used to induce oil accumulation in oleaginous yeast as part of a global stress response. While research has demonstrated that post-translational modifications (PTMs), including phosphorylation and protein cysteine thiol oxidation (redox PTMs), are involved in signaling pathways that regulate stress responses in metazoa and algae, their role in oleaginous yeast remain understudied and unexplored. Results Towards linking the yeast oleaginous phenotype to protein function, we integrated lipidomics, redox proteomics, and phosphoproteomics to investigate Rhodotorula toruloides under nitrogen-rich and starved conditions over time. Our lipidomics results unearthed interactions involving sphingolipids and cardiolipins with ER stress and mitophagy. Our redox and phosphoproteomics data highlighted the roles of the AMPK, TOR, and calcium signaling pathways in regulation of lipogenesis, autophagy, and oxidative stress response. As a first, we also demonstrated that lipogenic enzymes including fatty acid synthase are modified as a consequence of shifts in cellular redox states due to nutrient availability. Conclusions We conclude that lipid accumulation is largely a consequence of carbon rerouting and autophagy governed by changes to PTMs, and not increases in the abundance of enzymes involved in central carbon metabolism and fatty acid biosynthesis. Our systems-level approach sets the stage for acquiring multidimensional data sets for protein structural modeling and predicting the functional relevance of PTMs using Artificial Intelligence/Machine Learning (AI/ML). Coupled to those bioinformatics approaches, the putative PTM switches that we delineate will enable advanced metabolic engineering strategies to decouple lipid accumulation from nitrogen limitation.
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spelling doaj-art-9690da07a92f431ab6b40bf0cd5c8f6d2025-08-20T04:01:52ZengBMCBiotechnology for Biofuels and Bioproducts2731-36542025-07-0118112310.1186/s13068-025-02657-yNitrogen limitation causes a seismic shift in redox state and phosphorylation of proteins implicated in carbon flux and lipidome remodeling in Rhodotorula toruloidesAustin Gluth0Jeffrey J. Czajka1Xiaolu Li2Kent J. Bloodsworth3Josie G. Eder4Jennifer E. Kyle5Rosalie K. Chu6Bin Yang7Wei-Jun Qian8Pavlo Bohutskyi9Tong Zhang10Biological Sciences Division, Pacific Northwest National LaboratoryEnergy and Environment Directorate, Pacific Northwest National LaboratoryBiological Sciences Division, Pacific Northwest National LaboratoryBiological Sciences Division, Pacific Northwest National LaboratoryBiological Sciences Division, Pacific Northwest National LaboratoryBiological Sciences Division, Pacific Northwest National LaboratoryBiological Sciences Division, Pacific Northwest National LaboratoryDepartment of Biological Systems Engineering, Washington State UniversityBiological Sciences Division, Pacific Northwest National LaboratoryBiological Sciences Division, Pacific Northwest National LaboratoryBiological Sciences Division, Pacific Northwest National LaboratoryAbstract Background Oleaginous yeast are prodigious producers of oleochemicals, offering alternative and secure sources for applications in foodstuff, skincare, biofuels, and bioplastics. Nitrogen starvation is the primary strategy used to induce oil accumulation in oleaginous yeast as part of a global stress response. While research has demonstrated that post-translational modifications (PTMs), including phosphorylation and protein cysteine thiol oxidation (redox PTMs), are involved in signaling pathways that regulate stress responses in metazoa and algae, their role in oleaginous yeast remain understudied and unexplored. Results Towards linking the yeast oleaginous phenotype to protein function, we integrated lipidomics, redox proteomics, and phosphoproteomics to investigate Rhodotorula toruloides under nitrogen-rich and starved conditions over time. Our lipidomics results unearthed interactions involving sphingolipids and cardiolipins with ER stress and mitophagy. Our redox and phosphoproteomics data highlighted the roles of the AMPK, TOR, and calcium signaling pathways in regulation of lipogenesis, autophagy, and oxidative stress response. As a first, we also demonstrated that lipogenic enzymes including fatty acid synthase are modified as a consequence of shifts in cellular redox states due to nutrient availability. Conclusions We conclude that lipid accumulation is largely a consequence of carbon rerouting and autophagy governed by changes to PTMs, and not increases in the abundance of enzymes involved in central carbon metabolism and fatty acid biosynthesis. Our systems-level approach sets the stage for acquiring multidimensional data sets for protein structural modeling and predicting the functional relevance of PTMs using Artificial Intelligence/Machine Learning (AI/ML). Coupled to those bioinformatics approaches, the putative PTM switches that we delineate will enable advanced metabolic engineering strategies to decouple lipid accumulation from nitrogen limitation.https://doi.org/10.1186/s13068-025-02657-y
spellingShingle Austin Gluth
Jeffrey J. Czajka
Xiaolu Li
Kent J. Bloodsworth
Josie G. Eder
Jennifer E. Kyle
Rosalie K. Chu
Bin Yang
Wei-Jun Qian
Pavlo Bohutskyi
Tong Zhang
Nitrogen limitation causes a seismic shift in redox state and phosphorylation of proteins implicated in carbon flux and lipidome remodeling in Rhodotorula toruloides
Biotechnology for Biofuels and Bioproducts
title Nitrogen limitation causes a seismic shift in redox state and phosphorylation of proteins implicated in carbon flux and lipidome remodeling in Rhodotorula toruloides
title_full Nitrogen limitation causes a seismic shift in redox state and phosphorylation of proteins implicated in carbon flux and lipidome remodeling in Rhodotorula toruloides
title_fullStr Nitrogen limitation causes a seismic shift in redox state and phosphorylation of proteins implicated in carbon flux and lipidome remodeling in Rhodotorula toruloides
title_full_unstemmed Nitrogen limitation causes a seismic shift in redox state and phosphorylation of proteins implicated in carbon flux and lipidome remodeling in Rhodotorula toruloides
title_short Nitrogen limitation causes a seismic shift in redox state and phosphorylation of proteins implicated in carbon flux and lipidome remodeling in Rhodotorula toruloides
title_sort nitrogen limitation causes a seismic shift in redox state and phosphorylation of proteins implicated in carbon flux and lipidome remodeling in rhodotorula toruloides
url https://doi.org/10.1186/s13068-025-02657-y
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