Trois, Cristina.Anandraj, Akash.White, Sarah Anne.2014-10-302014-10-3020142014http://hdl.handle.net/10413/11417Ph.D. University of KwaZulu-Natal, Durban 2014.Hydrogen is universally known as the most efficient renewable energy source capable of meeting global energy demands. Chlamydomonas reinhardtii has the ability to produce biohydrogen during the metabolic engineering of the photosynthetic pathways. The aim of this study was to 1) use leachate as a feedstock to enhance microalgal biomass and subsequent hydrogen production, 2) use Pulse Amplitude Modulated (PAM) Fluorometry to elucidate the role of photosystem one during hydrogen production, 3) use Nicotinamide Adenine Dinucleotide Phosphate (NADPH) fluorescence as an indicator of hydrogen production and 4) design a modular pilot scale biohydrogen bioprocessing system implementing experimental findings into a conceptual model. This resulted in a cost effective source of renewable hydrogen produced from waste. The use of 16% landfill leachate was found to increase biomass production by 26% as compared to using Tris- Acetate Phosphate (TAP) media alone. Hydrogen induction resulted in an increased gas synthesis of 37% as well as an increased production period of 8 days compared to the normal 5 days. Landfill leachate further reduced the costs as it acted as a free nutrient source with the added ecological advantage of leachate treatment. Hydrogen production was induced by sulphur depletion and physiological parameters were measured using PAM Fluorometry. Photosystem I was found to be dominant during hydrogen production while photosystem II was down-regulated due to the sulphur depletion and damaged D1 proteins. NADPH fluorescence was significantly correlated to hydrogen yields allowing for NADPH to be utilised as a molecular indicator for hydrogen synthesis. The overall functionality of this bioprocessing system relies on the optimum physiological functioning of cells. The above findings were implemented into a pilot scale design, maximising the physiological performance during hydrogen production. This study has contributed knowledge regarding the production of hydrogen gas from leachate, the physiological changes of photosystem I during hydrogen production and the use of NADPH fluorescence as an indicator. The fundamental theories of bioprocessing incorporate a firm understanding of cellular and biochemical processes. The use of molecular indicators determined from physiological studies can be used at pilot scale to improve overall efficiency of hydrogen production.en-ZALeachate.Synthesis gas.Biochemical engineering.Photobiology.Theses--Civil engineering.Thesis