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Evolutionary significance of an algal gene encoding an [FeFe]-hydrogenase with F-domain homology and hydrogenase activity in Chlorella variabilis NC64A

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Abstract

[FeFe]-hydrogenases (HYDA) link the production of molecular H2 to anaerobic metabolism in many green algae. Similar to Chlamydomonas reinhardtii, Chlorella variabilis NC64A (Trebouxiophyceae, Chlorophyta) exhibits [FeFe]-hydrogenase (HYDA) activity during anoxia. In contrast to C. reinhardtii and other chlorophycean algae, which contain hydrogenases with only the HYDA active site (H-cluster), C. variabilis NC64A is the only known green alga containing HYDA genes encoding accessory FeS cluster-binding domains (F-cluster). cDNA sequencing confirmed the presence of F-cluster HYDA1 mRNA transcripts, and identified deviations from the in silico splicing models. We show that HYDA activity in C. variabilis NC64A is coupled to anoxic photosynthetic electron transport (PSII linked, as well as PSII-independent) and dark fermentation. We also show that the in vivo H2-photoproduction activity observed is as O2 sensitive as in C. reinhardtii. The two C. variabilis NC64A HYDA sequences are similar to homologs found in more deeply branching bacteria (Thermotogales), diatoms, and heterotrophic flagellates, suggesting that an F-cluster HYDA is the ancestral enzyme in algae. Phylogenetic analysis indicates that the algal HYDA H-cluster domains are monophyletic, suggesting that they share a common origin, and evolved from a single ancestral F-cluster HYDA. Furthermore, phylogenetic reconstruction indicates that the multiple algal HYDA paralogs are the result of gene duplication events that occurred independently within each algal lineage. Collectively, comparative genomic, physiological, and phylogenetic analyses of the C. variabilis NC64A hydrogenase has provided new insights into the molecular evolution and diversity of algal [FeFe]-hydrogenases.

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Abbreviations

LED:

Light emitting diode

MBBM:

Modified Bold’s Basal Medium

MOPS:

3-Morpholinopropanesulfonic acid

MTAP:

Modified tris-acetate-phosphate

MV:

Methyl viologen

PAR:

Photosynthetically active radiation

PSII/PSI:

Photosystem II/Photosystem I

TAP:

Tris-acetate-phosphate

HYDA:

[FeFe]-hydrogenase

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Acknowledgments

The authors of this work gratefully acknowledge the United States Air Force Office of Scientific Research under grant FA9550-05-1-0365 (to JEM, ESB, GA, DK, GCD, JWP, MCP), a National Aeronautics and Space Administration Graduate Research Program (GSRP) grant #NNG05GL52H (JEM, MCP), the Chemical Sciences, Geosciences and Biosciences Division, Office of Basic Energy Sciences, Office of Science, U.S. Department of Energy (Contract No. DE-FG02-96ER20225) for funding the development of extracellular metabolite quantitation (NMUM, MLG), the National Aeronautics and Space Administration Astrobiology Institute under grant NNA08C-N85A [Astrobiology Biogeocatalysis Research Center (ESB, JWP)], and a National Aeronautics and Space Administration Astrobiology Institute postdoctoral fellowship (ESB). The work conducted by the United States Department of Energy Joint Genome Institute is supported by the Office of Science of the United States Department of Energy under Contract No. DE-AC02-05CH11231. Thanks are extended to Dr. James Gurnon and Dr. James L. Van Etten (University of Nebraska) for providing C. variabilis NC64A and access to transcriptome data. We also acknowledge exceptional technical assistance from Edward Dempsey (Chemistry Dept. Colorado School of Mines).

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  1. Devin Karns

    Present address: Department of Mechanical Engineering, University of Washington, Seattle, WA, 98195, USA

Authors and Affiliations

  1. Department of Chemistry and Geochemistry, Colorado School of Mines, Golden, CO, 80401, USA

    Randor Radakovits & Matthew C. Posewitz

  2. Division of Environmental Science and Engineering, Colorado School of Mines, Golden, CO, 80401, USA

    Jonathan E. Meuser

  3. Department of Chemistry and Biochemistry and the Astrobiology Biogeocatalysis Research Center, Montana State University, Bozeman, MT, 59717, USA

    Eric S. Boyd & John W. Peters

  4. Department of Chemistry and Chemical Biology and the Waksman Institute of Microbiology, Rutgers, The State University of NJ, Piscataway, NJ, 08854-8020, USA

    Gennady Ananyev & G. Charles Dismukes

  5. Department of Engineering, Colorado School of Mines, Golden, CO, 80401, USA

    Devin Karns

  6. Chemical and Biosciences Center, National Renewable Energy Laboratory, Golden, CO, 80401-3393, USA

    U. M. Narayana Murthy & Maria L. Ghirardi

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Meuser, J.E., Boyd, E.S., Ananyev, G. et al. Evolutionary significance of an algal gene encoding an [FeFe]-hydrogenase with F-domain homology and hydrogenase activity in Chlorella variabilis NC64A. Planta 234, 829–843 (2011). https://doi.org/10.1007/s00425-011-1431-y

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