Abstract

The need for energy and the associated burden are ever growing. It is crucial to develop new technologies for generating clean and efficient energy for society to avoid upcoming energetic and environmental crises. Sunlight is the most abundant source of energy on the planet. Consequently, it has captured our interest. Certain microalgae possess the ability to capture solar energy and transfer it to the energy carrier, H2. H2 is a valuable fuel, because its combustion produces only one by-product: water. However, the establishment of an efficient biophotolytic H2 production system is hindered by three main obstacles: (1) the hydrogen-evolving enzyme, [FeFe]-hydrogenase, is highly sensitive to oxygen; (2) energy conversion efficiencies are not economically viable; and (3) hydrogen-producing organisms are sensitive to stressful conditions in large-scale production systems. This study aimed to circumvent the oxygen sensitivity of this process with a cyclic hydrogen production system. This approach required a mutant that responded to high temperatures by reducing oxygen evolution. To that end, we randomly mutagenized the green microalgae, Chlamydomonas reinhardtii, to generate mutants that exhibited temperature-sensitive photoautotrophic growth. The selected mutants were further characterized by their ability to evolve oxygen and hydrogen at 25 and 37 °C. We identified four candidate mutants for this project. We characterized these mutants with PSII fluorescence, P700 absorbance, and immunoblotting analyses. Finally, we demonstrated that these mutants could function in a prototype hydrogen-producing bioreactor. These mutant microalgae represent a novel approach for sustained hydrogen production.

Highlights

  • Photosynthesis is one of the most important life-sustaining reactions on our planet

  • Oxygenic photosynthesis was limited by a shortage of oxidized electron acceptors, which are plentiful on the current earth surface

  • Colonies (n = 12,000) grown from single cells were screened for temperature-sensitive photoautotrophic (TSP) mutants

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Summary

Introduction

Photosynthesis is one of the most important life-sustaining reactions on our planet It provides the energy required for the survival of all life forms, and it underlies the accumulation of fossil fuels, the main source of energy for sustaining modern human lifestyles (Barber 2004; Nelson and Yocum 2006). Oxygenic photosynthesis, which evolved shortly thereafter, used light energy and water to produce reducing equivalents. These equivalents could be used as chemical energy, and oxygen evolved as a by-product. About 1.5 billion years passed before metal compounds in the oceans and on the earth’s surface were oxidized, which allowed oxygen to accumulate in the atmosphere (Nelson 2011).

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