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Metabolically Engineered Rhodobacter sphaeroides RV strains for Improved Biohydrogen Photoproduction Combined with Disposal of Food Wastes

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Abstract

Three differently metabolically engineered strains, 2 single PHA- and Hup- mutants and one double PHA-/Hup- mutant, of the purple nonsulfur photosynthetic bacterium Rhodobacter sphaeroides RV, were constructed to improve a light-driven biohydrogen production process combined with the disposal of solid food wastes. These phenotypes were designed to abolish, singly or in combination, the competition of H2 photoproduction with polyhydroxyalkanoate (PHA) accumulation by inactivating PHA synthase activity, and with H2 recycling by abolishing the uptake hydrogenase enzyme. The performance of these mutants was compared with that of the wild-type strain in laboratory tests carried out in continuously fed photobioreactors using as substrates both synthetic media containing lactic acid and media from the acidogenic fermentation of actual fruit and vegetable wastes, containing mainly lactic acid, smaller amounts of acetic acia, and traces of higher volatile acids. With the lactic acid-based synthetic medium, the single Hup- and the double PHA-/Hup- mutants, but not the single PHA- mutant, exhibited increased rates of H2 photoproduction, about one third higher than that of the wild-type strain. With the food-waste-derived growth medium, only the single Hup- mutant showed higher rates of H2 production, but all 3 mutants sustained a longer-term H2 photoproduction phase than the wild-type strain, with the double mutant exhibiting overall the largest amount of H2 evolved. This work demonstrates the feasibility of single and multiple gene engineering of microorganisms to redirect their metabolism for improving H2 photoproduction using actual waste-derived substrates.

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Acknowledgments

We thank Dr. John Benemann for critical discussions and many helpful suggestions during the preparation of this publication. This work was performed under the management of the Research Institute of Innovative Technology for the Earth (RITE) as part of the Research and Development Project on Environmentally Friendly Technology for the Production of Hydrogen supported by New Energy and Industrial Technology Development Organization (NEDO), Japan.

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  1. EniTecnologie S.p.A., Environmental Technology Research Center, Via F. Maritano, 26-20097, San Donato Milanese, Milan, Italy

    Elisabetta Franchi, Claudio Tosi, Giuseppe Scolla, Gino Della Penna, Francesco Rodriguez & Paola Maria Pedroni

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  1. Elisabetta Franchi
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  2. Claudio Tosi
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  3. Giuseppe Scolla
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  6. Paola Maria Pedroni
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Correspondence to Paola Maria Pedroni.

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Franchi, E., Tosi, C., Scolla, G. et al. Metabolically Engineered Rhodobacter sphaeroides RV strains for Improved Biohydrogen Photoproduction Combined with Disposal of Food Wastes. Mar Biotechnol 6, 552–565 (2004). https://doi.org/10.1007/s10126-004-1007-y

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