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The effects of pretreatment method of cow dung compost, which was employed as natural hydrogen bacteria source, on the microbial community, population distribution of microbes and hydrogen production potential were investigated in the batch tests. The maximum hydrogen yield of 290.8 mL/L-culture appeared in the pretreated method A (infrared drying) by dark fermentation. The pretreated method of compost significantly affected microbial succession, population distribution of microbes. Both Clostridium sp. and Enterobacter sp. were found to be two species of preponderant hydrogen-producing bacteria, the next best was Bacteroides sp. and Veillonella sp., the last was Lactobacillus sp. and Streptococcus sp., which were also essential. The results showed that the mutualism and symbiosis relations of the mixed bacteria played a critical role in hydrogen fermentation process.

Effects of pretreatment of natural bacterial source and raw material on biohydrogen production in fermentative biohydrogen production process were investigated systematically. Biohydrogen production from stale corn slurry was found to be feasible and effective by A1 (water soak) pretreated compost and B1 (gelatinization) pretreated stale corn. The results were further confirmed by the verification test with working volume of 8 L, in which the maximum hydrogen yield of 262 mL H 2 /g-substrate, hydrogen production rate of 39 mL/g h −1 and the corresponding hydrogen content of 50% was observed at fixed substrate concentration of 10 g/L, working pH 5.0–5.5 and 36 ± 1 °C. The effluent was mostly composed of acetate and butyrate. Subsequently, two new hydrogen producing strains were isolated from the effluent sludge in the running bioreactor, and they were preliminarily identified as Clostridium and Enterobacter , respectively, according to the routine screening examinations.

The key process parameters of solid state enzymolysis for the generation of soluble sugar (SS) and bio-hydrogen production from corn stalk were optimized by the response surface methodology (RSM) based on a three factor-five level central composite design (CCD), respectively. The result showed that the optimal solid state enzymolysis condition from corn stalk was 47.7 °C, SCED of 0.054 g/g and 10.3 days for the maximum SS yield of 526 mg/g-TVS. Correspondingly, the optimal enzymolysis conditions from corn stalk appeared at 46.3 °C, SCED of 0.049 g/g and 7.5 days for the maximum hydrogen yield of 205.5 mL/g-TVS from the hydrolyzed substrate by the next dark fermentation. In addition, the bio-hydrogen production mechanism from corn stalk was preliminary investigated by XRD and SEM analyses. The results suggested that the solid state enzymolysis of substrate played a vital role in the effective conversion of corn stalk into bio-hydrogen by dark fermentation.

A hydrogen producing strain F.P 01 was newly isolated from cow dung sludge in an anaerobic bioreactor. The strain F.P 01 was a mesophilic and facultative anaerobic bacterium, which exhibited gram-negative staining in both the exponential and stationary growth phases, and a regular long rod-shaped bacteria with the size of 0.6–0.9 μm × 1.2–2.5 μm, and also could biodegrade a variety of carbohydrates such as glucose, xylose, maltose, etc. The effects of important process parameters on hydrogen producing of F.P 01 were further investigated from hydrogen fermentation of maltose by strain F.P 01, including substrate concentration, medium pH, etc. And the results showed that hydrogen production potential and hydrogen production rate from maltose of this strain F.P 01 was180 mLH 2 /g-maltose and 4.0 mLH 2 /h, respectively. The corresponding hydrogen concentration of 58–73% was also be observed. Both butyric acid and acetic acid as main by-product was left in the reactor.

Biohydrogen production from corncob using natural anaerobic microflora was reported for the first time. The optimum pretreatment condition for the corncob was determined to be 100 °C, 30 min, and 1% HCl (w/w). The maximum hydrogen yield of 107.9 ml/g-TVS and hydrogen production rate of 4.20 ml/g-TVS h −1 was obtained under the condition of 10 g/l substrate concentration and initial pH 8.0. Butyrate and acetate were the dominant metabolic by-products of hydrogen fermentation. Chemical composition analysis, Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) were used to study the mechanism of degrading corncob for hydrogen production. The amorphous domains of cellulose and hemicellulose were hydrolyzed into fermentable saccharides through acid pretreatment and the microorganisms had a devastating effect on the crystallinity of the cellulose. The hydrogen yield from pretreated corncob was much higher than from raw corncob. Therefore, the acid pretreatment played a crucial role on hydrogen production from corncob.

Strategies were adopted to cost-efficiently produce cellulose-hydrogen by anaerobic fermentation in this paper. First, cellulase used for hydrolyzing cellulose was prepared by solid-state fermentation (SSF) on cheap biomass from Trichoderma viride. Several cultural conditions for cellulase production on cheap biomass such as moisture content, inoculum size and culture time were studied. And the components of solid-state medium were optimized using statistical methods to further improve cellulase capability. Second, the crude cellulase was applied to cellulose-hydrogen process directly. The maximal hydrogen yield of 122 ml/g-TVS was obtained at the substrate concentration of 20 g/L and cultured time of 53 h. The value was about 45-fold than that of raw corn stalk wastes. The hydrogen content in the biogas was 44–57%(v/v) and there was no significant methane gas observed.

A hydrogen producing strain newly isolated from anaerobic sludge in an anaerobic bioreactor, was identified as Clostridium beijerinckii Fanp3 by 16S rDNA gene sequence analysis and detection by BioMerieux Vitek. The strain could utilize various carbon and nitrogen sources to produce hydrogen, which indicates that it has the potential of converting renewable wastes into hydrogen. In batch cultivations, the optimal initial pH of the culture medium was between 6.47 and 6.98. Using 0.15 M phosphate as buffer could alleviate the medium acidification and improve the overall performance of C. beijerinckii Fanp3 in hydrogen production. Culture temperature of 35 °C was established to be the most favorable for maximum rate of hydrogen production. The distribution of soluble metabolic products (SMP) was also greatly affected by temperature. Considering glucose as a substrate, the activation energy ( E a ) for hydrogen production was calculated as 81.01 kcal/mol and 21.4% of substrate energy was recovered in the form of hydrogen. The maximal hydrogen yield and the hydrogen production rate were obtained as 2.52 mol/mol-glucose and 39.0 ml/g-glucose h −1 , respectively. These results indicate that C. beijerinckii Fanp3 is an ideal candidate for the fermentative hydrogen production.