This paper reports on the optimum conditions for simultaneous hydrogen and butyric acid production from microalgae (Chlorella sp.) using enriched anaerobic mixed cultures as inoculum. The fermentation was objectively carried out under acidogenic conditions to achieve butyric acid for further ABE fermentation in solventogenesis stage. The main effects of initial pH (5 and 7), temperature (35 °C and 55 °C), and substrate concentration (40, 60, 80, and 100 g-VS/L) for hydrogen and butyric acid production were evaluated by using batch fermentation experiment. The major effects on hydrogen and butyric acid production are pH and temperature. The highest production of hydrogen and butyric acid was observed at pH 7 and temperature 35 °C. Using initial Chlorella sp. concentration of 80 g-VS/L or 100 g-VS/L at pH 7 and temperature 35 °C could produce hydrogen with an average yield of 22 mL-H₂/g-VS along with high butyric acid production yield of 0.05 g/g-VS, suggesting that microalgae (Chlorella sp.) has potential to be converted directly to butyric acid by using acidogenesis under above optimum conditions.

Food waste is categorized as organic solid waste, which has a negative impact on environmental sustainability. Food waste was simultaneously used for the feasible generation of mixed volatile fatty acids (VFAs) and bio-hydrogen by deploying dark fermentation. Original anaerobic digested sludge was prepared via the shock technique with 50 g/L glucose under thermophilic temperature (55 °C). The pretreated inoculum was found capable of converting 10 g VS/L food waste to hydrogen with a rather high yield of 135.2 ± 7 mL H₂/VS_added. The effect of various concentrations of food waste, including 10.2, 16.3, 20.3, and 26.4 g VS/L, on mixed VFAs production was subsequently carried out in batch dark fermentation. The highest butyric acid concentration (5.26 ± 0.22 g/L) in soluble metabolites was obtained from batch dark fermentation with 26.4 g VS/L of food waste. The dominant Clostridium thermobutyricum, Clostridium sporogenes, and Octadecobacter sp. found in the batch of dark fermentation of food waste could confirm the effectiveness of the load shock pretreatment method for inoculum preparation. The continuous stirred tank reactor (CSTR) inoculated with mixed cultures, also prepared via the load shock pretreatment method and without the addition of external nutrients, was operated by feeding 26.4 g VS/L food waste at the kinetically designed HRT for 4 days, corresponding to an organic loading rate (OLR) of 7.6 g VS/L·d. Under steady state conditions, promising butyric acid (5.65 ± 0.51 g/L)-rich mixed VFAs were achieved along with the hydrogen yield of 104.9 ± 11.0 mL-H₂/g VS_added, which is similar to the upper side of the previously reported yields (8.8 ± 0.6–103.6 ± 0.6 mL-H₂/g VS).

Oil palm empty fruit bunches (EFB) and palm oil decanter cake (DC) were used to investigate biogas production by using solid-state anaerobic co-digestion (SS-AcoD) with 15% total solid (TS) content. Solid state anaerobic digestion (SS-AD) using substrate to inoculum (S:I) ratio of 3:1, methane yields of 353.0 mL-CH₄/g-VS and 101.5 mL-CH₄/g-VS were respectively achieved from mono-digestion of EFB without oil palm ash (OPA) addition and of DC with 10% OPA addition under mesophilic conditions 35 °C. By adding 5% OPA to SS-AD using 3:1 S:I ratio under thermophilic conditions (55 °C), mono-digestion of EFB and DC provided methane yields of 365.0 and 160.3 mL-CH₄/g-VS, respectively. Furthermore, SS-AcoD of EFB:DC at 1:1 mixing ratio (volatile solid, VS basis), corresponding to carbon to nitrogen (C:N) ratio of 32, gathering with S:I ratio of 3:1 and 5% ash addition, synergistic effect is observed together with similar methane yields of 414.4 and 399.3 mL-CH₄/g-VS, achieved under 35 °C and 55 °C, respectively. According to first order kinetic analysis under synergistic condition, methane production rate from thermophilic operation is 5 times higher than that from mesophilic operation. Therefore, SS-AcoD could be potentially beneficial to generate biogas from EFB and DC.