Hydrogen (H2) is considered as fuel for future and its biological production from renewable feedstocks is very promising. Dark fermentation of fruit-vegetable waste (FVW) and cottage cheese whey (CCW) for the production of H2 constitutes a promising way for combining energy generation and lignocellulosic waste management. In this work, process kinetics of biohydrogen production via dark fermentation of FVW and CCW by pretreated anaerobic sludge inocula were investigated. To inhibit H2 consuming methanogens, the effects of various inoculum pretreatment, viz., 2-bromoethanesulfonate, heat-shock, acid, alkali, UV, and ultrasonication on H2-production were investigated which revealed 2-bromoethanesulfonate, heat-shock and acid-treated inoculum resulted in maximum bioH2 production and yield of 118.12 ± 1.05, 93.37 ± 1.3, 96.32 mMol/L and 1.66, 1.22 ± 0.01, 1.39 ± 0.02 mMol/gCODinitial, respectively. The effects of system initial pH, substrate to inoculum (S0/X0) and carbon to nitrogen (C/N) ratio on H2-production were evaluated which revealed maximum H2 production and yield could be achieved at pH 7, S0/X0 of 10.6 gCOD/gVS, and C/N 26.8. Modified Gompertz model and Modified Logistic model were used to define various kinetic parameters pertaining to cumulative H2-production which showed high R2 values (≥0.98). The influence of pH on H2 and ethanol/volatile fatty acids production kinetics were evaluated using Andrew's and Ratkowsky's model showing relatively good R2 values (≥0.62). Remarkably high production of ethanol (2.43 ± 0.28 mg/L) was noticed alongside H2 production at pH 7 suggesting that bioethanol can be recovered at the end of fermentative H2 production. Terminal Restriction Fragment Length Polymorphism and 16s rDNA sequencing revealed dominance of 9 bacterial species such as Escherichia coli, Clostridium butyricum, Streptococcus henryi, and 6 others uncultured bacteroides. This research determined different kinetic parameters for an enhanced H2 production strategy by co-fermentation of FVW and CCW providing an understanding of process behavior which will in turn help in the upscaling of the H2-production processes.
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