Integrated Amino Acid Profiling and 4D-DIA Proteomics Reveal Protein Quality Divergence and Metabolic Adaptation in <i>Cordyceps</i> Species

To explore the differences in protein quality among classic medicinal entomopathogenic fungi and to evaluate their metabolic adaptability, we analyzed the amino acid composition and proteomic characteristics of <i>Cordyceps sinensis</i> (CS), <i>Cordyceps militaris</i> (CM),...

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Main Authors: Chuyu Tang, Yuejun Fan, Tao Wang, Jie Wang, Mengjun Xiao, Min He, Xiyun Chang, Yuling Li, Xiuzhang Li
Format: Article
Language:English
Published: MDPI AG 2025-05-01
Series:Journal of Fungi
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Online Access:https://www.mdpi.com/2309-608X/11/5/365
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author Chuyu Tang
Yuejun Fan
Tao Wang
Jie Wang
Mengjun Xiao
Min He
Xiyun Chang
Yuling Li
Xiuzhang Li
author_facet Chuyu Tang
Yuejun Fan
Tao Wang
Jie Wang
Mengjun Xiao
Min He
Xiyun Chang
Yuling Li
Xiuzhang Li
author_sort Chuyu Tang
collection DOAJ
description To explore the differences in protein quality among classic medicinal entomopathogenic fungi and to evaluate their metabolic adaptability, we analyzed the amino acid composition and proteomic characteristics of <i>Cordyceps sinensis</i> (CS), <i>Cordyceps militaris</i> (CM), and <i>Cordyceps cicadae</i> (CC). Quantitative analysis showed CM contained the highest crude protein and lysine, methionine, threonine, and valine. CS adapted to high-altitude hypoxia and exhibited lower protein but elevated leucine, isoleucine, and histidine contents, which may contribute to membrane stabilization and oxidative stress resistance. CC displayed higher non-essential amino acids such as arginine, proline, and tyrosine, reflecting active nitrogen metabolism. Four-dimensional data-independent acquisition (4D-DIA) proteomics identified 495 differentially expressed proteins (DEPs). Compared with CS, CM and CC displayed upregulated glutamate oxaloacetate transaminases 2 (GOT2), glutamate dehydrogenase (GDH), and argininosuccinate synthase 1 (ASS1) coordinately regulate nitrogen flux through the alanine-aspartate-glutamate metabolic network and urea cycle, supporting metabolic intermediate replenishment for energy metabolism. The upregulation of branched-chain keto acid dehydrogenase E1 subunit alpha (BCKDHA) and acyl-CoA dehydrogenase short/branched chain (ACADSB) in CM and CC facilitated the integration of branched-chain amino acid catabolism with the TCA cycle, explaining species-specific differences in protein content. This study presents the first application of 4D-DIA proteomics to compare CS, CM, and CC, providing insights into quality divergence mechanisms in medicinal fungi.
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spelling doaj-art-c285627038b24253a5b1fc2679e5c0c92025-08-20T01:56:16ZengMDPI AGJournal of Fungi2309-608X2025-05-0111536510.3390/jof11050365Integrated Amino Acid Profiling and 4D-DIA Proteomics Reveal Protein Quality Divergence and Metabolic Adaptation in <i>Cordyceps</i> SpeciesChuyu Tang0Yuejun Fan1Tao Wang2Jie Wang3Mengjun Xiao4Min He5Xiyun Chang6Yuling Li7Xiuzhang Li8State Key Laboratory of Plateau Ecology and Agriculture, Qinghai Academy of Animal and Veterinary Sciences, Qinghai University, Xining 810016, ChinaState Key Laboratory of Plateau Ecology and Agriculture, Qinghai Academy of Animal and Veterinary Sciences, Qinghai University, Xining 810016, ChinaState Key Laboratory of Plateau Ecology and Agriculture, Qinghai Academy of Animal and Veterinary Sciences, Qinghai University, Xining 810016, ChinaState Key Laboratory of Plateau Ecology and Agriculture, Qinghai Academy of Animal and Veterinary Sciences, Qinghai University, Xining 810016, ChinaState Key Laboratory of Plateau Ecology and Agriculture, Qinghai Academy of Animal and Veterinary Sciences, Qinghai University, Xining 810016, ChinaState Key Laboratory of Plateau Ecology and Agriculture, Qinghai Academy of Animal and Veterinary Sciences, Qinghai University, Xining 810016, ChinaThe Department of Pharmacy, Qinghai Institute of Health Sciences, Xining 810016, ChinaState Key Laboratory of Plateau Ecology and Agriculture, Qinghai Academy of Animal and Veterinary Sciences, Qinghai University, Xining 810016, ChinaState Key Laboratory of Plateau Ecology and Agriculture, Qinghai Academy of Animal and Veterinary Sciences, Qinghai University, Xining 810016, ChinaTo explore the differences in protein quality among classic medicinal entomopathogenic fungi and to evaluate their metabolic adaptability, we analyzed the amino acid composition and proteomic characteristics of <i>Cordyceps sinensis</i> (CS), <i>Cordyceps militaris</i> (CM), and <i>Cordyceps cicadae</i> (CC). Quantitative analysis showed CM contained the highest crude protein and lysine, methionine, threonine, and valine. CS adapted to high-altitude hypoxia and exhibited lower protein but elevated leucine, isoleucine, and histidine contents, which may contribute to membrane stabilization and oxidative stress resistance. CC displayed higher non-essential amino acids such as arginine, proline, and tyrosine, reflecting active nitrogen metabolism. Four-dimensional data-independent acquisition (4D-DIA) proteomics identified 495 differentially expressed proteins (DEPs). Compared with CS, CM and CC displayed upregulated glutamate oxaloacetate transaminases 2 (GOT2), glutamate dehydrogenase (GDH), and argininosuccinate synthase 1 (ASS1) coordinately regulate nitrogen flux through the alanine-aspartate-glutamate metabolic network and urea cycle, supporting metabolic intermediate replenishment for energy metabolism. The upregulation of branched-chain keto acid dehydrogenase E1 subunit alpha (BCKDHA) and acyl-CoA dehydrogenase short/branched chain (ACADSB) in CM and CC facilitated the integration of branched-chain amino acid catabolism with the TCA cycle, explaining species-specific differences in protein content. This study presents the first application of 4D-DIA proteomics to compare CS, CM, and CC, providing insights into quality divergence mechanisms in medicinal fungi.https://www.mdpi.com/2309-608X/11/5/365<i>Cordyceps sinensis</i><i>Cordyceps militaris</i><i>Cordyceps cicadae</i>4D-DIA proteomicamino acid
spellingShingle Chuyu Tang
Yuejun Fan
Tao Wang
Jie Wang
Mengjun Xiao
Min He
Xiyun Chang
Yuling Li
Xiuzhang Li
Integrated Amino Acid Profiling and 4D-DIA Proteomics Reveal Protein Quality Divergence and Metabolic Adaptation in <i>Cordyceps</i> Species
Journal of Fungi
<i>Cordyceps sinensis</i>
<i>Cordyceps militaris</i>
<i>Cordyceps cicadae</i>
4D-DIA proteomic
amino acid
title Integrated Amino Acid Profiling and 4D-DIA Proteomics Reveal Protein Quality Divergence and Metabolic Adaptation in <i>Cordyceps</i> Species
title_full Integrated Amino Acid Profiling and 4D-DIA Proteomics Reveal Protein Quality Divergence and Metabolic Adaptation in <i>Cordyceps</i> Species
title_fullStr Integrated Amino Acid Profiling and 4D-DIA Proteomics Reveal Protein Quality Divergence and Metabolic Adaptation in <i>Cordyceps</i> Species
title_full_unstemmed Integrated Amino Acid Profiling and 4D-DIA Proteomics Reveal Protein Quality Divergence and Metabolic Adaptation in <i>Cordyceps</i> Species
title_short Integrated Amino Acid Profiling and 4D-DIA Proteomics Reveal Protein Quality Divergence and Metabolic Adaptation in <i>Cordyceps</i> Species
title_sort integrated amino acid profiling and 4d dia proteomics reveal protein quality divergence and metabolic adaptation in i cordyceps i species
topic <i>Cordyceps sinensis</i>
<i>Cordyceps militaris</i>
<i>Cordyceps cicadae</i>
4D-DIA proteomic
amino acid
url https://www.mdpi.com/2309-608X/11/5/365
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