Enhanced hydrogen evolution was pursued in this work. Rhodobacter sp. (Rb) and Rhodopseudomonas palustris (Rp), single or mixed were used to extract hydrogen molecules from Chlorella fusca biomass. To elevate their fermentable contents, Chlorella was grown at nitrogen and/or phosphorus deprivation. Besides, cellulase and/or macerozyme, Triton X100 or sonicated yeast were applied for further biohydrogen fermentation. Utilizing hydrolysates of mineral deprived Chlorella cultures, Rb exhibited relatively higher cumulative hydrogen (4200 ml L1 ) than Rp (2500 ml L1 ) while mixed cultures attained significantly higher levels (4700 ml L1 ). Triton or enzymes significantly enhanced hydrogen evolution, with more effectiveness of macerozyme than cellulase. A novel use of sonicated yeast, as enzymes pool, induced the highest significant collective H2 (up to 47 times that of microalgal supernatant). Sonicated yeast induced a remarkable hydrolysis of algae, as inferred from increased reducing sugars. However, hydrogen evolution efficiency exhibited poor proportionality with reducing sugars, indicating fermentation of other metabolites. © 2021 Hydrogen Energy Publications LLC. Published by Elsevier Ltd. All rights reserved