a b s t r a c tThis paper discusses the overall energy consumption and greenhouse gas emissions when extracting pigments and producing hydrogen from Spirogyra sp. microalga biomass. The energy evaluation from the biomass leftovers was also included in this work. The influence of the functional unit and different allocation criteria on the biorefinery assessments is also shown. The study consists of laboratory tests showing Spirogyra sp. growth, harvesting, drying, pigment extraction and fermentation by Clostridium butyricum. Electrocoagulation and solar drying were tested and compared to conventional centrifugation and electrical dewatering in terms of their energy consumption for harvesting and dewatering, respectively. To discuss the biorefinery viability, the pigments and biohydrogen (bioH 2 ) retail costs are considered against operational costs according to electricity needs. The low yield of biochemical hydrogen and the high energy requirements for the pigment extraction were identified as main topics for further research. This research hopefully contributes to highlight the importance of energy and emission balances in order to decide on feasibility of the biorefinery.

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a b s t r a c tThe biological hydrogen production from Spirogyra sp. biomass was studied in a SBR (sequential batch reactor) equipped with a biogas collecting and storage system. Two acid hydrolysis pre-treatments (1N and 2N H 2 SO 4 ) were applied to the Spirogyra biomass and the subsequent fermentation by Clostridium butyricum DSM 10702 was compared. The 1N and 2N hydrolyzates contained 37.2 and 40.8 g/L of total sugars, respectively, and small amounts of furfural and HMF (hydroxymethylfurfural). These compounds did not inhibit the hydrogen production from crude Spirogyra hydrolyzates. The fermentation was scaled up to a batch operated bioreactor coupled with a collecting system that enabled the subsequent characterization and storage of the biogas produced. The cumulative hydrogen production was similar for both 1N and 2N hydrolyzate, but the hydrogen production rates were 438 and 288 mL/L.h, respectively, suggesting that the 1N hydrolyzate was more suitable for sequential batch fermentation. The SBR with 1N hydrolyzate was operated continuously for 13.5 h in three consecutive batches and the overall hydrogen production rate and yield reached 324 mL/L.h and 2.59 mol/mol, respectively. This corresponds to a potential daily production of 10.4 L H 2 /L Spirogyra hydrolyzate, demonstrating the excellent capability of C. butyricum to produce hydrogen from microalgal biomass.

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