Improving the yield from fermentative hydrogen production

Efforts to increase H₂ yields from fermentative H₂ production include heat treatment of the inoculum, dissolved gas removal, and varying the organic loading rate. Although heat treatment kills methanogens and selects for spore-forming bacteria, the available evidence indicates H₂ yields are not maximized compared to bromoethanesulfonate, iodopropane, or perchloric acid pre-treatments and spore-forming acetogens are not killed. Operational controls (low pH, short solids retention time) can replace heat treatment. Gas sparging increases H₂ yields compared to un-sparged reactors, but no relationship exists between the sparging rate and H₂ yield. Lower sparging rates may improve the H₂ yield with less energy input and product dilution. The reasons why sparging improves H₂ yields are unknown, but recent measurements of dissolved H₂ concentrations during sparging suggest the assumption of decreased inhibition of the H₂-producing enzymes is unlikely. Significant disagreement exists over the effect of organic loading rate (OLR); some studies show relatively higher OLRs improve H₂ yield while others show the opposite. Discovering the reasons for higher H₂ yields during dissolved gas removal and changes in OLR will help improve H₂ yields.     

Animal digestive strategies versus anaerobic digestion bioprocesses for biogas production from lignocellulosic biomass

Herbivorous mammals and wood-eating insects are fairly effective in the digestion of plant polymers, such as lignocellulosics. In order to improve methane production from the lignocellulosic biomass, several kinds of anaerobic digestion processes derived from animal models have been devised. However, the rates of biodegradation occurring in the anaerobic bioreactors still remain lower than in animal guts. The effectiveness of the digestive systems of those animals results from the concerted action of the various enzymes (e.g. cellulases, xylanases, esterases, ligninases) produced in their guts as well as their integration with mechanical and chemical actions. Powerful pretreatment (prefermentation) operations are integrated to and support efficiently the microbial fermentation system, e.g. the rumination (i.e. mechanical) in ruminants and the secretion of endogenous cellulases (i.e. enzymatic) or the alkaline treatment (chemical) at mid-way in xylophagous insects. The oxygen gradients along the gastrointestinal tract may also stimulate the hydrolytic activities of some microbial populations. In addition, the solid retention time, the digesta flow and the removal of the end-products are well ordered to enable animals to thrive on a complex polymer such as lignocellulose. At the same time, technologies were developed to degrade lignocellulosic biomass, such as the rumen derived anaerobic digestion (RUDAD) process and the rumen simulating technique (RUSITEC), more elaborated and using rumen microbial consortia. Overall, they showed that the fermentation taking place in the rumen fermentation and even in the hindgut are biological processes that go beyond the limited environmental conditions generally found in anaerobic digesters. Hence, knowledge on herbivores' digestion mechanisms might be better exploited in the design and operation of anaerobic digesters. This literature review is a cross-analysis of the relevant information about the digestive strategies of herbivorous and wood-eating animals and the bioengineering techniques in lignocelluloses degradation. The aim is to highlight strategies of animals' digestion simulation for more effective anaerobic digestion of lignocellulosic compounds and other solid residues.     

Energy analysis of ethanol production from sweet sorghum

The Piedmont System is a collection of equipment for efficiently removing the juice from sweet sorghum stalks for the production of ethanol. The concept is to separate the whole stalks into pith and rind-leaf fractions, pass only the pith fraction through a screw press, and thus achieve an improvement in juice-expression efficiency and press capacity. An energy analysis was done for two options of this proposed harvesting/processing system: (Option 1) The juice is evaporated to syrup and used throughout the year to produce ethanol, and the by-products are used as cattle feed. (Option 2) The juice is fermented as it is harvested, and the by-products (along with other cellulosic materials) are used as feedstock for the remainder of the year. Energy ratios (energy output/energy input) of 0·9, 1·1 and 0·8 were found for sweet sorghum Option 1, sweet sorghum Option 2, and corn, respectively, as feedstocks for ethanol. If only liquid fuels are considered, the ratios are increased to 3·5, 7·9 and 4·5.     

Phycoremediation coupled production of algal biomass,harvesting and anaerobic digestion: Possibilities and challenges

Biogas produced from anaerobic digestion is a versatile and environment friendly fuel which traditionally utilizes cattle dung as the substrate. In the recent years, owing to its high content of biodegradable compounds, algal biomass has emerged as a potential feedstock for biogas production. Moreover, the ability of algae to treat wastewater and fix CO₂ from waste gas streams makes it an environmental friendly and economically feasible feedstock. The present review focuses on the possibility of utilizing wastewater as the nutrient and waste gases as the CO₂ source for algal biomass production and subsequent biogas generation. Studies describing the various harvesting methods of algal biomass as well as its anaerobic digestion have been compiled and discussed. Studies targeting the most recent advancements on biogas enrichment by algae have been discussed. Apart from highlighting the various advantages of utilizing algal biomass for biogas production, limitations of the process such as cell wall resistivity towards digestion and inhibitions caused due to ammonia toxicity and the possible strategies for overcoming the same have been reviewed. The studies compiled in the present review indicate that if the challenges posed in translating the lab scale studies on phycoremediation and biogas production to pilot scale are overcome, algal biogas could become the sustainable and economically feasible source of renewable energy.     

Stirring and biomass starter influences the anaerobic digestion of different substrates for biogas production

Here, we present the results of lab‐scale experiments conducted in a batch mode to determine the biogas yield of lipid‐rich waste and corn silage under the effect of stirring. Further semi‐continuous experiments were carried out for the lipid‐rich waste with/without stirring. Additionally, it was analyzed how the starter used for the batch experiment influences the digestion process. The results showed a significant stirring effect on the anaerobic digestion only when seed sludge from a biogas plant was used as a starter. In this case, the experiments without stirring yielded only about 50% of the expected biogas for the investigated substrates. The addition of manure slurry to the batch reactor as part of the starter improved the biogas production. The more diluted media in the reactor allowed a better contact between the bacteria and the substrates making stirring not necessary.     

Characterization of the biomass of the stems of sweet sorghum

In this study the amount of glucose, sucrose and fructose was determined in the water soluble fraction while cellulose, hemicellulose, pectin and lignin contents were determined in the alcohol insoluble fraction after hydrolysis. Stalks of sweet sorghum (Sorghum vulgare L., var. saccharatum) cv. Vespa, Soave, Roce and MN 1500 at the physiological ripeness stage were used. The results of the analysis of variance with the least significant difference method (LSD, = 0.05) show that cv. Vespa and Roce have a significantly higher total amount of glucose, fructose and sucrose and at the same time, a lower cellulose, pectin, hemicellulose and lignin content then cv. Soave and cv. MN 1500.     

Changes in bacterial community during fermentative hydrogen and acid production from organic waste by thermophilic anaerobic microflora

AIMS: Changes in fermentation pattern during the treatment of organic wastes containing solid materials by thermophilic anaerobic microflora were investigated with respect to product formation and bacterial community structure during hydrogen production. METHODS AND RESULTS: Anaerobic microflora enriched from sludge compost was cultivated using artificial garbage slurry in a continuous flow-stirred tank reactor. Product formation varied depending on pH and hydraulic retention time (HRT) applied. Community analysis by terminal restriction fragment length polymorphism and clone library analysis of polymerase chain reaction-amplified bacterial 16S rDNA indicated that difference in the fermentative product distribution could be caused by different populations of micro-organisms in the microflora. CONCLUSION: Hydrogen fermentation with acetate/butyrate formation was optimized at <1.0 d HRT at pH 5.0 and 6.0. Thermoanaerobacterium thermosaccharolyticum was the dominant hydrogen-producing micro-organism. Conversely, unidentified organisms became dominant after 4.0 d HRT at pH 7.0 and 8.0, where relatively high-solubilization efficiency of solid materials was observed with no production of hydrogen. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report describing product formation in the fermentation of solid organic wastes by a mixed population of micro-organisms. Various fermentation patterns including hydrogen fermentation were characterized and evaluated from engineering and microbial aspects.     

Growth media in anaerobic fermentative processes: The underestimated potential of thermophilic fermentation and anaerobic digestion

Fermentation and anaerobic digestion of organic waste and wastewater is broadly studied and applied. Despite widely available results and data for these processes, comparison of the generated results in literature is difficult. Not only due to the used variety of process conditions, but also because of the many different growth media that are used. Composition of growth media can influence biogas production (rates) and lead to process instability during anaerobic digestion. To be able to compare results of the different studies reported, and to ensure nutrient limitation is not influencing observations ascribed to process dynamics and/or reaction kinetics, a standard protocol for creating a defined growth medium for anaerobic digestion and mixed culture fermentation is proposed. This paper explains the role(s) of the different macro- and micronutrients, as well as the choices for a growth medium formulation strategy. In addition, the differences in nutrient requirements between mesophilic and thermophilic systems are discussed as well as the importance of specific trace metals regarding specific conversion routes and the possible supplementary requirement of vitamins. The paper will also give some insight into the bio-availability and toxicity of trace metals. A remarkable finding is that mesophilic and thermophilic enzymes are quite comparable at their optimum temperatures. This has consequences for the trace metal requirements of thermophiles under certain conditions. Under non-limiting conditions, the trace metal requirement of thermophilic systems is about 3 times higher than for mesophilic systems.     

Improvement of fermentative hydrogen production: various approaches

Fermentation of biomass or carbohydrate-based substrates presents a promising route of biological hydrogen production compared with photosynthetic or chemical routes. Pure substrates, including glucose, starch and cellulose, as well as different organic waste materials can be used for hydrogen fermentation. Among a large number of microbial species, strict anaerobes and facultative anaerobic chemoheterotrophs, such as clostridia and enteric bacteria, are efficient producers of hydrogen. Despite having a higher evolution rate of hydrogen, the yield of hydrogen [mol H₂ (mol substrate^–1)] from fermentative processes is lower than that achieved using other methods; thus, the process is not economically viable in its present form. The pathways and experimental evidence cited in the literature reveal that a maximum of four mol of hydrogen can be obtained from substrates such as glucose. Modifications of the fermentation process, by redirection of metabolic pathways, gas sparging and maintaining a low partial pressure of hydrogen to make the reaction thermodynamically favorable, efficient product removal, optimum bioreactor design and integrating fermentative process with that of photosynthesis, are some of the ways that have been attempted to improve hydrogen productivity. This review briefly describes recent advances in these approaches towards improvement of hydrogen yield by fermentation.     

Modeling and optimization of fermentative hydrogen production

Biohydrogen is a sustainable energy resource due to its potentially higher efficiency of conversion to usable power, non-polluting nature and high energy density. The purpose of modeling and optimization is to improve, analyze and predict biohydrogen production. Biohydrogen production depends on a number of variables, including pH, temperature, substrate concentration and nutrient availability, among others. Mathematical modeling of several distinct processes such as kinetics of microbial growth and products formation, steady state behavior of organic substrate along with its utilization and inhibition have been presented. Present paper summarizes the experimental design methods used to investigate effects of various factors on fermentative hydrogen production, including one-factor-at-a-time design, full factorial and fractional factorial designs. Each design method is briefly outlined, followed by the introduction of its analysis. In addition, the applications of artificial neural network, genetic algorithm, principal component analysis and optimization process using desirability function have also been highlighted.     

Illumination enhances methane production from thermophilic anaerobic digestion

Incandescent lamp illumination enhanced methane production from a thermophilic anaerobic digestion reactor (55°C) supplied with glucose. After 10 days of operation, the volume of methane produced from light reactors was approximately 2.5 times higher than that from dark reactors. A comparison of the carbon balance between light and dark conditions showed that methane produced from hydrogen and carbon dioxide in the light reactors was higher than that from the dark reactors. When hydrogen or acetate was fed into the reactors, methane production with added hydrogen was faster and higher under light conditions than under dark conditions. The use of blue light-emitting diodes also enhanced methane production over that under dark conditions. The 16S rRNA gene copy numbers for Methanothermobacter spp. in the light reactor and in the dark reactor were at the same level. The copy number for Methanosarcina spp. in the light reactors was approximately double than that in the dark reactors. These results suggest that blue light enhances the methanogenic activity of hydrogenotrophic methanogens.     

Structured modeling of the anaerobic digestion of biomass particulates

Anaerobic digestion of biological organic particulates to methane has been described by a structured mathematical model based on multiple-reaction stoichiometry, conventional material balances, and liquid phase equilibrium chemistry. A general stoichiometric treatment for any set of multiple biological reactions is derived based on a unit mass of oxggen equivalents of the reactions limiting substrate. The model agrees well with two existing experimental studies of anaerobic digestion of biomass particulates. Hypothetical computer simulations are presented to illustrate possible instabilities of the anaerobic process under various operating scenarios.     

Batch and continuous fermentative production of hydrogen with anaerobic sludge entrapped in a composite polymeric matrix

Cell immobilization techniques were adopted to biohydrogen production using immobilized anaerobic sludge as the seed culture. Sucrose-based synthetic wastewater was converted to H₂ using batch and continuous cultures. A novel composite polymeric material comprising polymethyl methacrylate (PMMA), collagen, and activated carbon was used to entrap biomass for H₂ production. Using the PMMA immobilized cells, the favorable conditions for batch H₂ fermentation were 35 °C, pH 6.0, and an 20 g COD l⁻¹ of sucrose, giving a H₂ production rate of 238 ml h⁻¹ l⁻¹ and a H₂ yield of 2.25 mol H₂ mol sucrose⁻¹. Under these optimal conditions, continuous H₂ fermentation was conducted at a hydraulic retention time (HRT) of 4–8 h, giving the best H₂-producing rate of 1.8 l h⁻¹ l⁻¹ (over seven-fold of the best batch result) at a HRT of 6 h and a H₂ yield of 2.0 mol H₂ mol sucrose⁻¹. The sucrose conversion was essentially over 90% in all runs. The biogas consisted of only H₂ and CO₂. The major soluble metabolites were butyric acid, acetic acid, and 2,3-butandiol, while a small amount of ethanol also detected. The PMMA-immobilized-cell system developed in this work seems to be a promising H₂-producing process due to the high stability in continuous operations and the capability of achieving a competitively high H₂ production rate under a relatively low organic loading rate.     

Microbial and enzymatic improvement of anaerobic digestion of waste biomass

Metabolic activities of different microorganisms (Bacillus subtilis, B. licheniformis and Aspergillus niger) and hydrolytic enzymes (concentrations: 1 to 200 mg enzyme solids g^–1 feed) were studied individually and in combinations with respect to H₂ and methane production from damaged wheat grains. Bacillus subtilis, B. licheniformis and pre-existing hydrogen producers (control) produced 45 to 64 l H₂ kg^–1 total solids and subsequently, with the help of added methanogens, 155 to 220 l methane kg^–1 total solids could be produced. H₂ production from damaged wheat grains could be decreased to 28% or enhanced up to 152% with respect to control, by employing various microbial and enzymatic treatments. Similarly, it has been made possible to vary methane production capacities from as low as 17% to as high as 110% with respect to control.     

生物质制氢技术研究进展

氢能以其清洁,来源广泛及用途广等优点成为最有希望的替代能源之一,用可再生能源制氢是氢能发展的必然趋势。由于生物质制氢具有一系列独特的优点,它已成为发展氢经济颇具前景的研究领域之一。生物质制氢技术可以分为两类,一类是以生物质为原料利用热物理化学方法制取氢气,如生物质气化制氢,超临界转化制氢,高温分解制氢等热化学发制氢,以及基于生物质的甲烷、甲醇、乙醇的化学重整转化制氢等;另一类是利用生物转化途径转换制氢,包括直接生物光解,间接生物光解,光发酵,光合异养细菌水气转移反应合成氢气,暗发酵和微生物燃料电池等技术。本文综述了目前主要的生物质制氢技术及其发展概况,并分析了各技术的发展趋势。     

Fermentative hydrogen production from pretreated biomass: A comparative study

The aim of this work was to evaluate the potential of employing biomass resources from different origin as feedstocks for fermentative hydrogen production. Mild-acid pretreated and hydrolysed barley straw (BS) and corn stalk (CS), hydrolysed barley grains (BG) and corn grains (CG), and sugar beet extract (SB) were comparatively evaluated for fermentative hydrogen production. Pretreatments and/or enzymatic hydrolysis led to 27, 37, 56, 74 and 45 g soluble sugars/100 g dry BS, CS, BG, CG and SB, respectively. A rapid test was applied to evaluate the fermentability of the hydrolysates and SB extract. The thermophilic bacterium Caldicellulosiruptor saccharolyticus showed high hydrogen production on hydrolysates of mild-acid pretreated BS, hydrolysates of BG and CG, and SB extract. Mild-acid pretreated CS showed limited fermentability, which was partially due to inhibitory products released in the hydrolysates, implying the need for the employment of a milder pretreatment method. The difference in the fermentability of BS and CS is in strong contrast to the similarity of the composition of these two feedstocks. The importance of performing fermentability tests to determine the suitability of a feedstock for hydrogen production was confirmed.     

Metabolic flux network and analysis of fermentative hydrogen production

Fermentative hydrogen production (FHP) has received a great R & D interest in recent decades, as it offers a potential means of producing H₂ from a variety of renewable resources, even wastewater via a low energy continuous process. Various extracellular metabolites including ethanol, acetate, butyrate and lactate can be produced during the fermentation, building a complex metabolic network of the FHP. Except for the recognition of its complexity, the metabolic flux network has not been well understood. Studies on biochemical reactions and metabolic flux network associated with the FHP in anaerobic fermentation system have only been drawn attention in recent years. This review summarizes the biochemical reactions taking place in the metabolic network of FHP. We discuss how the key operation factors influence metabolism in the FHP process. Recently developed and applied technologies for metabolic flux analysis have been described. Future studies on the metabolic network to enhance fermentative hydrogen production by strict anaerobes are recommended. It is expected that this review can provide useful information in terms of fundamental knowledge and update technology for scientists and research engineers in the field of biological hydrogen production.