Modeling dark fermentation for biohydrogen production: ADM1-based model vs. Gompertz model

Biohydrogen production by dark fermentation in batch reactors was modeled using the Gompertz equation and a model based on Anaerobic Digestion Model (ADM1). The ADM1 framework, which has been well accepted for modeling methane production by anaerobic digestion, was modified in this study for modeling hydrogen production. Experimental hydrogen production data from eight reactor configurations varying in pressure conditions, temperature, type and concentration of substrate, inocula source, and stirring conditions were used to evaluate the predictive abilities of the two modeling approaches. Although the quality of fit between the measured and fitted hydrogen evolution by the Gompertz equation was high in all the eight reactor configurations with r² ∼0.98, each configuration required a different set of model parameters, negating its utility as a general approach to predict hydrogen evolution. On the other hand, the ADM1-based model (ADM1BM) with predefined parameters was able to predict COD, cumulative hydrogen production, as well as volatile fatty acids production, albeit at a slightly lower quality of fit. Agreement between the experimental temporal hydrogen evolution data and the ADM1BM predictions was statistically significant with r² > 0.91 and p-value <1E-04. Sensitivity analysis of the validated model revealed that hydrogen production was sensitive to only six parameters in the ADM1BM.     

Inhibitory effect of ethanol,acetic acid,propionic acid and butyric acid on fermentative hydrogen production

The inhibitory effect of added ethanol, acetic acid, propionic acid and butyric acid on fermentative hydrogen production by mixed cultures was investigated in batch tests using glucose as substrate. The experimental results showed that, at 35 °C and initial pH 7.0, during the fermentative hydrogen production, the substrate degradation efficiency, hydrogen production potential, hydrogen yield and hydrogen production rate all trended to decrease with increasing added ethanol, acetic acid, propionic acid and butyric acid concentration from 0 to 300 mmol/L. The inhibitory effect of added ethanol on fermentative hydrogen production was smaller than those of added acetic acid, propionic acid and butyric acid. The modified Han–Levenspiel model could describe the inhibitory effects of added ethanol, acetic acid, propionic acid and butyric acid on fermentative hydrogen production rate in this study successfully. The modified Logistic model could describe the progress of cumulative hydrogen production.     

Kinetic models for fermentative hydrogen production: A review

The kinetic models were developed and applied for fermentative hydrogen production. They were used to describe the progress of a batch fermentative hydrogen production process, to investigate the effects of substrate concentration, inhibitor concentration, temperatures, pH, and dilution rates on the process of fermentative hydrogen production, and to establish the relationship among the substrate degradation rate, the hydrogen-producing bacteria growth rate and the product formation rate. This review showed that the modified Gompertz model was widely used to describe the progress of a batch fermentative hydrogen production process, while the Monod model was widely used to describe the effects of substrate concentration on the rates of substrate degradation, hydrogen-producing bacteria growth and hydrogen production. Arrhenius model was used a lot to describe the effects of temperature on fermentative hydrogen production, while modified Han–Levenspiel model was used to describe the effects of inhibitor concentration on fermentative hydrogen production. The Andrew model was used to describe the effects of H⁺ concentration on the specific hydrogen production rate, while the Luedeking–Piret model and its modified form were widely used to describe the relationship between the hydrogen-producing bacteria growth rate and the product formation rate. Finally, some suggestions for future work with these kinetic models were proposed.     

Combined effects of temperature and pH on biohydrogen production by anaerobic digested sludge

A full factorial design was conducted to investigate the combined effects of temperatures and initial pH on fermentative hydrogen production by mixed cultures in batch tests. The experimental results showed that the modified Logistic model can be used to describe the progress of cumulative hydrogen production in the batch tests of this study. The modified Ratkowsky model can be used to describe the combined effects of the temperatures and initial pH on the substrate degradation efficiency, hydrogen yield and average hydrogen production rate. The temperatures and initial pH had interactive impact on fermentative hydrogen production. The maximum substrate degradation efficiency, the maximum hydrogen yield and the maximum average hydrogen production rate was predicted at the temperature of 37.8 °C and the initial pH of 7.1, 37.4 °C and 6.9, and 38.2 °C and 7.2, respectively. In general, the optimal temperature for the fermentative hydrogen production was around 37.8 °C and the optimal initial pH for the fermentative hydrogen production was around 7.1.     

Enhancing photo fermentative hydrogen production using ethanol rich dark fermentation effluents

The present study demonstrates the feasibility of a two-phase biorefinery process applied to waste substrates producing ethanol rich effluents. The process includes a dark fermentation step followed by photo fermentation and it is able to optimize hydrogen production from waste biomass. The study was conducted using winery wastewater as feedstock. The results indicate that no additional treatments are required when an appropriate dilution of the initial waste is applied. Microbial consortia contained in the winery wastewater promoted a fermentative ethanol pathway. The ethanol rich effluent was converted into hydrogen by phototrophic microorganisms. Despite the presence of inhibiting compounds, the adoption of a mixed phototrophic culture allowed to obtain good results in terms of hydrogen production. Specifically, up to 310 mLH₂ gCOD_consumed^?1 were obtained in the photo fermentative stage. The effectiveness of ethanol rich dark fermentation effluents for hydrogen production enhancement was demonstrated. Noteworthy, polyhydroxybutyrate was also produced during the experiments. The work faces two of the major challenges in the sequential dark fermentation and photo fermentation technology applied to real waste substrates: the minimization of pre-treatments and the enhancement of the hydrogen production yields using ethanol rich DFEs.     

Modeling of fermentative hydrogen production from the bacterium Ruminococcus albus: Definition of metabolism and kinetics during growth on glucose

The aim of the present study was to describe the fermentative pathway of Ruminococcus albus during hydrogen production from glucose by a quantitative kinetic model, taking into account the interactions among the metabolic products during their generation. Proper mathematical expressions were developed in order to adequately describe the microbial growth and metabolism of R. albus. For the estimation of kinetics constants of the process, the experimental data from batch experiments were simulated using a simplified and modified version of Anaerobic Digestion Model 1 on Aquasim as a modeling platform. Subsequently the accuracy of the model was verified by simulating the performance of a CSTR in four different hydraulic retention times. Batch experiments with different initial substrate concentrations and different initial hydrogen partial pressures were carried out in order to calculate the growth kinetics of the microorganism and investigate the effect of hydrogen partial pressure to the production of metabolites. Microbial growth was described using Monod kinetics, taking into account the inhibition at lower pH values as well as the substrate inhibition, and the metabolites' profile was described using suitable kinetic expressions. Acetate and ethanol production were assumed to occur simultaneously, by direct sugar consumption and the H₂ final yield was reversely connected to the accumulation of ethanol. Formate was considered to be produced by direct sugar consumption, and subsequently to break down to H₂ and CO₂. The degradation rate of formate, and consequently hydrogen production were shown to be influenced by hydrogen partial pressure.     

产氢菌的投加方式对强化发酵菌群产氢的影响

通过间歇培养研究了产氢菌Ethanoligenens sp B49的投加方式对生物制氢反应器的混合发酵菌群生物强化作用的影响.结果表明,产氢菌的投加方式对发酵菌群的产氢能力有显著影响.产氢菌发酵液的直接投加使发酵菌群的产氢能力下降,并引起培养液中发酵产物乙醇和乙酸浓度的显著增加.分析认为,产氢菌发酵液对发酵菌群的末端产物抑制和低pH值抑制作用是导致产氢作用受到抑制的主要原因.离心后单独投加产氢菌菌体可提高发酵菌群的产氢能力,起到强化产氢的作用.投加10.8%的产氢菌强化发酵菌群时,培养45h的累计产氢量为155.0 mL.比强化前发酵菌群培养的产氢量提高了21.5%.因此在利用产氢菌生物强化发酵菌群的研究中,应采用离心分离后单独投加产氢菌菌体的方式进行生物强化.     

Inoculum type response to different pHs on biohydrogen production from l-arabinose,a component of hemicellulosic biopolymers

Biohydrogen production from arabinose was examined using four different anaerobic sludges with different pHs ranging from 4.5 to 8.0. Arabinose (30 g l⁻¹) was used as the substrate for all experiments. Individual cumulative hydrogen production data was used to estimate the three parameters of the modified Gompertz equation. Higher hydrogen production potentials were observed for higher pH values for all the sludges. G2 (acclimated granular sludge) showed the highest hydrogen production potential and percentage of arabinose consumption compared to the other sludges tested. Granular sludges (G1 and G2) showed different behaviour than the suspended sludges (S1 and S2). The differences were observed to be smaller lag phases, the percentage of acetate produced, the higher percentage of ethanol produced, and the amount of arabinose consumed. A high correlation (R² = 0.973) was observed between the percentage of n-butyrate and the percentage of ethanol in G1 sludge, suggesting that ethanol/butyrate fermentation was the dominant fermentative pathway followed by this sludge. In S1, however, the percentage of n-butyrate was highly correlated with the percentage of acetate (R² = 0.980). This study indicates that granular sludge can be used for larger pH ranges without reducing its capacity to consume arabinose and achieve higher hydrogen production potentials.     

Optimization and kinetic modeling of an enhanced bio‐hydrogen fermentation with the addition of synergistic biochar and nickel nanoparticle

The improvement of hydrogen production was achieved by the addition of biochar (BC) and metal co‐factor nanoparticle Ni⁰ during the dark fermentation. A new hybrid approach by combing the artificial neural networks with the response surface methodology was applied to optimize the hydrogen production. The effects of operating conditions, ie, BC, metal cofactor Ni⁰, pH, and dosage of microbes, upon the hydrogen production together with the concentrations of other metabolites such as the acetic acid, propionic acid, butyric acid, and ethanol were extensively investigated. From kinetic study of the major metabolites, the acetate pathway was found to be apparently enhanced by the addition of synergistic factors. The modified anaerobic digestion model with the consideration of inhabitation factor was found to best represent the kinetics of hydrogen production and the formation of major metabolites.     

Modeling fermentative hydrogen production from sucrose supplemented with dairy manure

Our previous studies had shown that fermentative hydrogen production from sucrose could be improved with dairy manure as a supplement. In addition to contributing to nearly 10% more hydrogen yield at ambient temperature, dairy manure was shown to be capable of providing the required nutritional needs, buffering capacity, and hydrogen-producing organisms, improving the practical viability of fermentative hydrogen production. In this report, we present a kinetic model for fermentative hydrogen production from sucrose supplemented with dairy manure. This model includes hydrogen production from sucrose as well as from the soluble products hydrolyzed from particulate manure. The integrated model was calibrated using experimental data from one batch reactor and validated with dissolved COD, hydrogen, and volatile fatty acid data from four other reactors. Predictions by this model agreed well with the temporal trends in the experimental data, with r² averaging 0.85 for dissolved COD; 0.94 for total COD; 0.84 for hydrogen; 0.84 for acetic acid; and 0.89 for butyric acid; quality of fit in the case of propionic acid was lower with r² averaging 0.57.     

Comparison of metabolic pathway for hydrogen production in wild-type and mutant Clostridium tyrobutyricum strain based on metabolic flux analysis

There has been a great interest in fermentative hydrogen production during recent decades. However, the low H₂ yield associated with fermentative hydrogen production process continues to hinder its industrial application. It is delectable that a maximum 3.9 mol H₂ per mol glucose was obtained in fed-batch fermentation mode with a butyric acid over-producing Clostridium tyrobutyricum mutant, which to our knowledge is the highest H₂ yield ever got in the fermentation process with Clostridium sp. This study aimed to better understand the change of flux profile within the whole metabolic network and to conduct the metabolic flux analysis of fermentative hydrogen production. For the first time, we constructed a metabolic flux model for the anaerobic glucose metabolism of C. tyrobutyricum ATCC 25755, and revealed the internal mechanism responsible for the redistribution of the carbon flux in the mutant strain in comparison with the wide-type. The MFA methodology was used to study the fractional flux response to variations in operational pH, and revealed that pH was a significant operational parameter effecting on the fermentative hydrogen production process. Furthermore, the presence of NADH-ferredoxin oxidoreductase activity in this anaerobe was demonstrated. By measuring the activities of related enzymes in the biosynthesis pathway of hydrogen, we thus concluded that the increased specific activities of both NFOR and hydrogen-catalyzing enzyme (hydrogenase) would be attributed to the hydrogen over-producing.     

Application of a modified Anaerobic Digestion Model 1 version for fermentative hydrogen production from sweet sorghum extract by Ruminococcus albus

The aim of the present study was to evaluate the effectiveness of a developed, ADM1-based kinetic model for the hydrogen production process in batch and continuous cultures of the bacterium Ruminococcus albus grown on sweet sorghum extract as the sole carbon source. Although sorghum extract is known to contain at least two different sugars, i.e. sucrose and glucose, no biphasic growth was observed in batch cultures as such growth is reported to occur in cultures of R. albus with mixed substrates. Thus, taking into account that the main sugar of sweet sorghum extract is sucrose, batch experiments with different initial concentrations of sucrose were performed in order to estimate the growth kinetics of the bacterium on this substrate. The kinetic parameters used, concerning the endogenous metabolism of the bacterium as well as those concerning the effect of pH and hydrogen partial pressure (P_H2), were the same as those estimated in a previous study with glucose as carbon source. Subsequently, the experimental data of batch and continuous experiments with sweet sorghum extract were simulated based on the already developed, modified ADM1 model accounting for the use of sugar-based substrate. It was shown that the model which was developed on synthetic substrates was successful in adequately describing the behavior of the microorganism on a real substrate such as sweet sorghum extract and predicting the experimental results quite well with a deviation of the model predictions from the experimental results being between 5-18% for the hydrogen yield.     

Biohydrogen from complex carbohydrate wastes as feedstocks—Cellulose degraders from a unique series enrichment

Fermentative biohydrogen production is a particularly promising approach offering high hydrogen yields and fast rates. Its drawback is that it uses mono- and disaccharides for substrates and these are relatively expensive. However, more complex carbohydrates may be converted to the feedstocks for fermentative biohydrogen production using approaches derived from select organisms. Early data from a consortium with cellulolytic activity recovered from aerobic thermophilic swine waste product and sequentially selected with batch operation on distiller's dry grain (a product of dry grind ethanol manufacture) has been used as a source of inoculum. Mesophilic and thermophilic cellulose-degrading organisms have been isolated from this inoculum and are reported here. Work continues on further characterization of these organisms and their potential. This work will be considered in light of an organizational scheme of processes using aerobic or anaerobic as well as mesophilic or thermophilic organisms. Some literature examples will be discussed. It is possible that these organisms could be used in a saccharification/fermentation process using separate reactors or a single reactor with coimmobilized cells providing both aerobic saccharification and anaerobic hydrogen fermentation.     

Application of kinetic models in dark fermentative hydrogen production–A critical review

Kinetic modeling could be viewed as an important step in developing a bioprocess, since models can be used in process control, reducing costs and optimizing processes. In the present study, the application of kinetic models in dark fermentative hydrogen production has been investigated. A wide variety of kinetic models are addressed and compared regarding their accuracy to fit the data. This literature survey indicates that the modified Gompertz was extensively used to describe the production of hydrogen, organic acids and alcohols, substrate degradation, and biomass growth. The development of kinetic models can assist researchers to identify the most important variables, facilitate future research, and maximize hydrogen production.     

Continuous dark fermentative hydrogen production by mesophilic microflora: Principles and progress

Continuous, dark fermentative hydrogen production technology using mixed microflora at mesophilic temperatures may be suitable for commercial development. Clostridial-based cultures from natural sources have been widely used, but more information on the need for heat treatment of inocula and conditions leading to germination and sporulation are required. The amount of nutrients given in the literature vary widely. Hydrogen production is reported to proceed without methane production in the reactor in the pH range 4.5–6.7, with hydraulic retention times optimally between a few hours and 3 days depending on substrate. Higher substrate concentrations should be more energy-efficient but there are product inhibition limitations, for example from unionised butyric acid. Inhibition by H₂ can be reduced by stirring, sparging or extraction through membranes. Of the reactor types investigated, while granules have the best performance with soluble substrate, for particulate feedstock biofilm reactors or continuous stirred tank reactors may be most successful. A second stage is required to utilise the fermentation end products which, when cost-effective reactors are developed, may be photofermentation or microbial fuel cell technologies. Anaerobic digestion is a currently-available technology and the two-stage process is reported to give greater conversion efficiency than anaerobic digestion alone.     

Value analysis for commercialization of fermentative hydrogen production from biomass

In addition to producing hydrogen gas, biohydrogen production is also used to process wastewater. Therefore, this study specifically conducted value analyses of two different scenarios of fermentative hydrogen production from a biomass system: to increase the value of a wastewater treatment system and to specifically carry out hydrogen production. The analytical results showed that fermentative hydrogen production from a biomass system would increase the value of a wastewater treatment system and make its commercialization more feasible. In contrast, fermentative hydrogen production from a biomass system designed specifically for producing hydrogen gas would have a lower system value, which indicated that it is not yet ready for commercialization. The main obstacle to be overcome in promoting biohydrogen production technology and system application is the lack of sales channels for the system's products such as hydrogen gas and electricity. Thus, in order to realize its commercialization, this paper suggests that governments provide investment subsidies for the use of biohydrogen production technology and establish a buy-back tariff system for fuel cells.     

Metabolic flux analysis of hydrogen production network by Clostridium butyricum W5: Effect of pH and glucose concentrations

Fermentative hydrogen production by strict anaerobes has been widely reported. There is a lack of information related to metabolic flux distribution and its variation with respect to fermentation conditions in the metabolic production system. This study aimed to get a better understanding of the metabolic network and to conduct metabolic flux analysis (MFA) of fermentative hydrogen production by a recently isolated Clostridium butyricum strain W5. We chose the specific growth rate as the objective function and used specific H₂ production rate as the criterion to evaluate the experimental results with the in silico MFA. For the first time, we constructed an in silico metabolic flux model for the anaerobic glucose metabolism of C. butyricum W5 with assistance of a modeling program MetaFluxNet. The model was used to evaluate metabolic flux distribution in the fermentative hydrogen production network, and to study the fractional flux response to variations in initial glucose concentration and operational pH. The MFA results suggested that pH has a more significant effect on hydrogen production yield compared to the glucose concentration. The MFA is a useful tool to provide valuable information for optimization and design of the fermentative hydrogen production process.     

Clostridium species for fermentative hydrogen production: An overview

Dark fermentation is considered as a promising method for sustainable hydrogen generation. Microorganisms play a pivotal role in hydrogen production efficiency. Strains from genus Clostridium have been the most widely detected and used microorganisms in dark fermentative hydrogen production. In this review, the characteristics of hydrogen-producing Clostridium species are introduced, including the metabolic pathways, hydrogen production performance, and substrate diversity. In addition, strategies for improving the hydrogen production by Clostridium species were extensively reviewed, and the promising applications of the fermentation system from the points of economic and environmental benefits were displayed. Finally, perspectives concerning the further application of hydrogen production by Clostridium species were proposed. Extensive studies demonstrate that Clostridium species are good candidates for fermentative hydrogen production with both high hydrogen yield and wide substrate range, but more efforts are still needed to ensure the efficient and stable operation of the system, and make the process economically applicable.     

Dark fermentative biohydrogen production by mesophilic bacterial consortia isolated from riverbed sediments

Dark fermentative bacterial strains were isolated from riverbed sediments and investigated for hydrogen production. A series of batch experiments were conducted to study the effect of pH, substrate concentration and temperature on hydrogen production from a selected bacterial consortium, TERI BH05. Batch experiments for fermentative conversion of sucrose, starch, glucose, fructose, and xylose indicated that TERI BH05 effectively utilized all the five sugars to produce fermentative hydrogen. Glucose was the most preferred carbon source indicating highest hydrogen yields of 22.3 mmol/L. Acetic and butyric acid were the major soluble metabolites detected. Investigation on optimization of pH, temperature, and substrate concentration revealed that TERI BH05 produced maximum hydrogen at 37 °C, pH 6 with 8 g/L of glucose supplementation and maximum yield of hydrogen production observed was 2.0–2.3 mol H₂/mol glucose. Characterization of TERI BH05 revealed the presence of two different bacterial strains showing maximum homology to Clostridium butyricum and Clostridium bifermentans.     

Improving ADM1 model to simulate anaerobic digestion start-up with inhibition phase based on cattle slurry

The Anaerobic Digestion Model No.1 (ADM1) was improved to simulate an anaerobic digestion start-up phase. To improve the ADM1, a combined hydrolysis equation was used based on the Contois model of bacterial growth and the function of hydrolysis inhibition by VFA. The start-up with fresh cattle slurry was carried out in a pilot-scale reactor to calibrate the chosen parameters of the ADM1. The important aspects of model calibration were hydrolysis rate, the number of anaerobic microbes in cattle slurry, and the growth rate of bacteria. Good simulation results were achieved after calibration for the independent start-up test with pre-conditioned cattle slurry.