The production of molecular hydrogen (H2) by microalgae holds great promise, and immobilization techniques offer potential for further advancement in this field. The current study focuses on investigating the positive impact of immobilization on maintaining the stability and activity of photosystem II (PSII) over incubation time, with the aim of enhancing H2 production potential in green microalgae Chlamydomonas reinhardtii. For this purpose, immobilized cells within alginate beads were treated with small concentrations of 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB) inhibitor to induce the partial inhibition of inter-photosystem electron transport, recently reported as a novel approach for sustaining microalgal H2 production. A comparative analysis of fluorescence decay kinetic changes and EPR spectroscopy of the cell beads revealed the superior capacity of immobilization for sustaining PSII stability and activity in batch culture over time. Treatment of the cell beads with 3.5 μM DBMIB led to sustained H2 production yielding over 200 μmol H2/mg Chl within 3 weeks, with an average H2 production rate of approximately 10 μmol/mg Chl per day, both of which were roughly twice as high as those observed in free cells treated with DBMIB. Our findings underscore the significance of integrating immobilization with a proven and effective method for H2 production, thereby enhancing its sustainability and productivity.
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