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Investigating Cellular Responses During Photohydrogen Production by the Marine Microalga Tetraselmis subcordiformis by Quantitative Proteome Analysis

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Abstract

The marine microalga Tetraselmis subcordiformis could photoproduce hydrogen under the regulation of carbonyl cyanide m-chlorophenylhydrazone (CCCP), and a hydrogen production process kinetic analysis was characterized by two peaks, suggesting that two distinct mechanisms might exist in this alga. Therefore, 2D nanoliquid chromatography−tandem mass spectrometry (LC-MS/MS) was introduced to analyze the proteome of samples from different time points. A total of 912 proteins were identified, providing a global view of the cellular responses at the proteomic level. These proteins can be divided into multiple functional groups including stress responses, energy metabolism and redox homeostasis. The quantitative proteomic data provided more details on the electron donors for hydrogen production. During the first stage, photosystem II produced electrons for hydrogen production; during the second stage, metabolites were the major electron donors via nonphotochemical plastoquinone reduction by NADH dehydrogenase.

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Acknowledgments

This work was supported by the Ningbo Natural Science Foundation of China (2011A610028).

Compliance with Ethical Standards

This article does not contain any studies with human participants or animals performed by any of the authors.

Conflict of Interest

The authors declare that they have no conflict of interest.

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    Authors and Affiliations

    1. School of Food Science and Technology, Dalian Polytechnic University, Dalian, 116034, People’s Republic of China

      Chaofan Ji

    2. National Engineering Research Center of Seafood, Dalian, 116034, People’s Republic of China

      Chaofan Ji

    3. Marine Bioengineering Group, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, People’s Republic of China

      Xupeng Cao, Changhong Yao & Song Xue

    4. Novel Technology and Materials for Separation and Detection of Biomolecules Group, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, People’s Republic of China

      Hongwei Liu & Hanfa Zou

    5. Department of Biology and Pharmacy, Zhejiang Pharmaceutical College, 315100, Ningbo, People’s Republic of China

      Junge Qu

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    1. Chaofan Ji
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    3. Hongwei Liu
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    Corresponding author

    Correspondence to Song Xue.

    Additional information

    Chaofan Ji and Xupeng Cao contributed equally to this work.

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    Fig. S1

    Relative abundance of photosystem (A) and Calvin cycle (B) proteins from T. subcordiformis. Relative ratios of differentially expressed proteins were calculated between samples from S1-S3 and control. The value equals to fold change of the protein expression,which was calculated as log2(ratio). The enzyme designations are: PetA for apocytochrome f; PetB for cytochrome b6; PetC for cytochrome b6-f complex iron-sulfur subunit; PetD for cytochrome b6-f complex subunit 4; FNR for ferredoxin--NADP+ reductase; ATPF0A-1G for different F-type H+-transporting ATPase subunits; PsaA-L for different photosystem I subunits; PsbA-R for different photosystem II subunits; ALT for alanine transaminase; ASP for aspartate aminotransferase; PBP for fructose-1,6-bisphosphatase; FBA for fructose-bisphosphate aldolase; GAPDH for glyceraldehyde 3-phosphate dehydrogenase; MDH for malate dehydrogenase; PGK for phosphoglycerate kinase; prkB for phosphoribulokinase; PK for pyruvate kinase; PPDK for pyruvate, orthophosphate dikinase; RPIA for ribose 5-phosphate isomerase A; RBCL/S for ribulose-bisphosphate carboxylase large/small chain; RPE for ribulose-phosphate 3-epimerase; SBPASE for sedoheptulose-bisphosphatase; TKTL for transketolase; TIM for triosephosphate isomerase. (GIF 3 kb)

    High resolution image (TIFF 901 kb)

    Fig. S2

    Relative abundance of starch metabolism (A) and glycolysis (B) proteins from T. subcordiformis. The enzyme designations are: AGL for 4-alpha-glucanotransferase; AGPase for glucose-1-phosphate adenylyltransferase; amyA for alpha-amylase; FK for fructokinase; GK for glucokinase; GPI for glucose-6-phosphate isomerase; PGM for phosphoglucomutase; PYG for starch phosphorylase; SS for starch synthase; ACSS for acetyl-CoA synthetase; ADH for alcohol dehydrogenase; DLD for dihydrolipoamide dehydrogenase; ENO for enolase; FBA for fructose-bisphosphate aldolase; G6PE for glucose-6-phosphate 1-epimerase; NQR for NADPH2:quinone reductase; PFK for 6-phosphofructokinase; PGAM for phosphoglycerate mutase. (GIF 2 kb)

    High resolution image (TIFF 992 kb)

    Fig. S3

    Relative abundance of fermentation and TCA proteins from T. subcordiformis. The enzyme designations are: DLST for dihydrolipoamide succinyltransferase; PC for pyruvate carboxylase; SDH for succinate dehydrogenase; PFL for formate C-acetyltransferase; OGDH for 2-oxoglutarate dehydrogenase; DLD for dihydrolipoamide dehydrogenase; CS for citrate synthase; ACO for aconitate hydratase; LSC for succinyl-CoA synthetase. (GIF 1 kb)

    High resolution image (TIFF 496 kb)

    Fig. S4

    Relative abundance of oxidative phosphorylation (A) and redox related (B) proteins from T. subcordiformis. The enzyme designations are: ATP(e)F for F-type H+-transporting ATPase subunits; PPA for inorganic pyrophosphatase; SEPHS for selenide, water dikinase; UQCRFS for ubiquinol-cytochrome c reductase; ATPeV for V-type H+-transporting ATPase subunit; FNR for ferredoxin--NADP+ reductase; GRX for glutaredoxin; PRX for peroxiredoxin; TRX for thioredoxin; APR for adenylyl-sulfate reductase; gshB for glutathione synthase; GSH for hydroxyacylglutathione hydrolase; ECM4 for putative glutathione S-transferase; GST for glutathione S-transferase; GPX for glutathione peroxidase; glutathione reductase for GR; NDH for NADH dehydrogenase; NDUFS for NADH dehydrogenase (ubiquinone) Fe-S protein; NDUFV for NADH dehydrogenase (ubiquinone) flavoprotein; NDUFA/B for NADH dehydrogenase (ubiquinone) 1 alpha/beta subcomplex. (GIF 3 kb)

    High resolution image (TIFF 885 kb)

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    Ji, C., Cao, X., Liu, H. et al. Investigating Cellular Responses During Photohydrogen Production by the Marine Microalga Tetraselmis subcordiformis by Quantitative Proteome Analysis. Appl Biochem Biotechnol 177, 649–661 (2015). https://doi.org/10.1007/s12010-015-1769-x

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