Evaluation of ultrasonication as a treatment strategy for enhancement of biohydrogen production from complex distillery wastewater and process optimization

Dark fermentation of distillery wastewater (DWW) gives a lower hydrogen yield (HY) and hydrogen production rate (R), owing to the complexity and a recalcitrant nature of effluent. Therefore, an effective pretreatment of DWW becomes imperative for the improvement of biohydrogen production. In the present study, the efficacy of ultrasonic pretreatment for enhancement of biohydrogen production from DWW was evaluated in batch test. Several variables, such as COD input, ultrasonic density (UD), and ultrasonication time (UT) were studied for optimization using response surface methodology integrated with desirability function. The highest HY, 10.95 mmol/g COD, and R, 6.67 mmol/L h, were obtained for batch test of ultrasonically pretreated DWW under optimal conditions for COD, UD and UT at 56 g/L, 0.12 W/mL, and 17 min, respectively. The significant relative enhancement of HY, 101%, and R, 103%, implies that ultrasonically treated DWW is about 1.2–1.4 times more effective for enhanced biohydrogen production from complex DWW compared to unsonicated DWW.     

Ferric oxide/date seed activated carbon nanocomposites mediated dark fermentation of date fruit wastes for enriched biohydrogen production

Biohydrogen production from biomass waste, not only addresses the energy demand in a renewable manner but also resolves the safe disposal issues associated with these biowastes. Also, scalable and low-cost techniques to enhance biohydrogen production have gained more attraction and are highly explored. In this research work, date-palm fruit wastes have been studied for their biohydrogen production potential using Enterobacter aerogenes by dark fermentation. Hydrogen yield and productivity were improved through the addition of iron oxide nanoparticles (Fe₃O₄ NPs) and its date seed activated carbon nanocomposites (Fe₃O₄/DSAC) to the fermentation media. Studies on discrete inclusions of these NPs showed that the appropriate dosage of NPs promoted, while higher dosages repressed the hydrogen production performance. Optimal dosage and fermentation time was observed as 150 mg/L and 24 h for both the additives. Fe₃O₄/DSAC nanocomposites showed better hydrogen production enhancement than Fe₃O₄ NPs. Maximum hydrogen yield of 238.7 mL/g was obtained for the 150 mg/L nanocomposites, which was 65.7% higher than that of the standalone Fe₃O₄ NPs and three folds higher than the yield of the control run without any NPs inclusion (78.4 mL/g). Metabolites analysis showed that the hydrogen evolution followed the ethanol-acetate pathway. Formation levels of longer chain propionate and butyrate co-metabolites were significantly low in the presence of Fe₃O₄/DSAC than Fe₃O₄. The carbon support in the nanocomposites acted as an adsorbent-buffer, which favored the medium pH in-addition to the stimulatory effects of Fe₃O₄ NPs. Cell growth and specific hydrogenase activity analysis were also performed to supplement the hydrogen production results. Gompertz and modified Logistic kinetic models were employed for kinetic modeling of experimental hydrogen production values. The Fe₃O₄/DSAC nanocomposites exhibited significant application potential for the production of biohydrogen from date fruit wastes.     

Whey waste as potential feedstock for biohydrogen production

In-house isolate Clostridium sp. IODB-O3 was exploited for biohydrogen production using cheese whey waste in batch fermentation. Analysis of cheese whey shows, it is enriched with lactose, lactic acid and protein components which were observed most favourable for biohydrogen production. Biohydrogen yield by IODB-O3 was compared with the cultures naturally occurring in waste solely or in combinations, and found that Clostridium sp. IODB-O3 was the best producer. The maximum biohydrogen yield obtained was 6.35 ± 0.2 mol-H₂/mol-lactose. The cumulative H₂ production (ml/L), 3330 ± 50, H₂ production rate (ml/L/h), 139 ± 5, and specific H₂ production (ml/g/h), 694 ± 10 were obtained. Clostridium sp. IODB-O3 exhibited better H₂ yield from cheese whey than the reported values in literature. Importantly, the enhancement of biohydrogen yield was observed possibly due to absence of inhibitory compounds, presence of essential nutrients, protein and lactic acid fractions which supported better cell growth than that of the lactose and glucose media. Carbon balance was carried out for the process which provided more insights in IODB-O3 metabolic pathway for biohydrogen production. This study may help for effective utilization of whey wastes for economic large scale biohydrogen production.     

Dynamic modelling of Rhodopseudomonas palustris biohydrogen production: Perturbation analysis and photobioreactor upscaling

Developing kinetic models to simulate Rhodopseudomonas palustris biohydrogen production within different configurations of photobioreactors (PBRs) poses a significant challenge. In this study, two types of PBRs: schott bottle-based and vertical tubular-based, were investigated, and three original contributions are presented. Firstly, a mechanistic model was constructed to simulate effects of light intensity, light attenuation and temperature on biomass growth and biohydrogen synthesis, previously not unified for photosynthetic bacteria. Secondly, perturbation analysis was exploited to identify critical parameters influencing the accuracy of the model. Thirdly, two parameters: effective light coefficient and biohydrogen enhancement coefficient, both linked to the PBR's transport phenomena were proposed for process scale-up prediction. By comparing against experimental data, the model's accuracy was confirmed to be high. Moreover, the enhancement of biohydrogen production rate by improved culture mixing and gas removal was also described mechanistically. This provides important advances for future efficient design of PBRs and process online optimisation.     

Determining the effect of trace elements on biohydrogen production from fruit and vegetable wastes

Trace elements are one of the important parameters for dark fermentative H₂ production because they work as co-factors in H₂ formation biochemistry. Lack or excess of trace element and its concentrations could be an important reason for the low yield of H₂ production. In this study, the effects of 11 different trace elements (Fe, Ni, Zn, Co, Cu, Mn, Al, B, Se, Mo and W) were tested at two levels in terms of biohydrogen production from Fruit and Vegetable Wastes (FVW) with Biochemical Hydrogen Potential (BHP) Tests using Plackett-Burman statistical design. 1.1–2.8 times enhancement of biohydrogen production was determined with its addition. The most effective trace elements were found as Zn and Ni. In order to reveal the resident microbial flora, Denaturing Gradient Gel Electrophoresis (DGGE) analysis was carried out on all BHP effluent samples. Results of DGGE analysis, four microbial sequences evaluated as Clostridium sp., Clostridium baratii, Uncultured bacterium, Uncultured Streptococcus sp., and their similarity rates were 99%, 100%, 89%, 98%, respectively.     

Optimization for simultaneous enhancement of biobutanol and biohydrogen production

Acetone-butanol-ethanol (ABE) fermentation guarantees a sustainable route for biohydrogen and biobutanol production. This research work is committed towards the enhancement of biohydrogen and biobutanol production by single and multi-parameter optimization for the improvement of substrate energy recovery using C. saccharoperbutylacetonicum. Single parameters optimization (SPO) manifested that headspace of 60% (v/v) and butyric acid supplementation of 9 g/L and temperatures of 30 °C and 37 °C were suitable for obtaining maximum biohydrogen and biobutanol production, respectively. The interaction between these parameters was further evaluated by implementing a 5-level 3-factor Central Composite Design (CCD). In the present study, a central composite design was employed to enhance the biohydrogen and biobutanol production. In addition, the experimental results were analyzed by response surface methodology (RSM) and artificial intelligence (AI) techniques such as artificial neural network (ANN). The prediction capability of RSM was further compared with ANN for predicting the optimum parameters that would lead to maximum biohydrogen and biobutanol production. ANN yielded higher values of biohydrogen and biobutanol. ANN was found to be superior as compared to RSM in terms of prediction accuracy for both biohydrogen and biobutanol because of its higher coefficient of determination (R²) and lower root mean square error (RMSE) value. Process temperature (32.65 °C), headspace (58.21% (v/v)) and butyric acid supplementation (9.16 g/L) led to maximum substrate energy recovery of 78% with biohydrogen and biobutanol production of 5.9 L/L and 16.75 g/L, respectively. Process parameter optimization led to a significant increase in substrate energy recovery from Biphasic fermentation.     

Application of nanotechnology in dark fermentation for enhanced biohydrogen production using inorganic nanoparticles

Fermentation is an important innovation by mankind and this process is used for converting organic substrate into useful products. Using natural conditions, specifically, light and dark conditions, photo-fermentation and dark fermentation techniques can be developed and operated under controlled conditions. Generally, products such as biofuels, bioactive compounds and enzymes have been produced using the dark fermentation method. However, the major requirement for today's industralized world is biofuels in its clean and pure forms. Biohydrogen is the most efficient and cleanest form of energy produced using dark fermentation of organic substrates. Nevertheless, the quantity of biohydrogen produced via dark fermentation is low. In order to increase the product quantity and quality, several internal and external stress or alterations are made to conventional fermentation conditions. In recent times, nanotechnology has been introduced to enhance the rate of dark fermentation. Nanoparticles (NPs), specifically, inorganic NPs such as silver, iron, titanium oxide and nickel have increased the production rate of biohydrogen. Therefore, the present review focuses on exploring the potential of nanotechnology in the dark fermentation of biohydrogen production, the mechanisms involved, substrates used and changes to be made to increase the production efficiency of dark fermentation.     

Nanoparticles augmentation on biogas yield from microalgal biomass anaerobic digestion

This study evaluates the influence of metal and metal-oxide nanoparticles (NPs) on biogas production from green microalgae Enteromorpha. The concentration of metallic NPs (Ni, Co) was 1 mg/L and oxides NPs (Fe₃O₄, MgO) was 10 mg/L. An anaerobic digestion was carried out batch-wise with working volume, operating temperature, mixing rate and hydraulic retention time as 500 ml, 37 °C, 150 rpm and 170 h, respectively. The measurements of chemical oxygen demand (COD), volatile fatty acids (VFAs), reducing sugar and biogas production were observed to monitor effectivity of nanoparticles. The results showed that NPs has moderate positive influence in biogas production until 60 h of retention time but significantly improve afterward. The maximum total biogas yield of 624 ml was achieved by Fe₃O₄ NPs whereas highest biohydrogen, 51.42% (v/v) was achieved by Ni NPs. The cumulative increase in biogas production for Fe₃O₄, Ni, Co and MgO NPs was 28%, 26%, 9% and 8%, respectively. A modified Gompertz and Logistic function model were used to determine kinetic constants of the reaction. The logistic model has the better predicting ability for microalgae anaerobic digestion.     

Biohydrogen production via integrated sequential fermentation using magnetite nanoparticles treated crude enzyme to hydrolyze sugarcane bagasse

This study presents a potential approach to enhance integrated sequential biohydrogen production from waste biomass using magnetite nanoparticle (Fe₃O₄ NPs) which is synthesized through waste seeds of Syzygium cumini. Consequences of 0.5% Fe₃O₄ NPs have been investigated on the thermal and pH stability of fungal crude cellulase. It is noticed that Fe₃O₄ NPs treated enzyme and control exhibits 100% activity in the temperature range of 45–60 °C and 45–55 °C, respectively. Moreover, Fe₃O₄ NPs treated enzyme showed extended thermal stability in the temperature range of 50–60 °C up to 12 h. Beside this, Fe₃O₄ NPs treated enzyme possesses 100% stability in the pH range of 5.0–7.0 whereas control exhibited only at pH 6.0. Enzymatic hydrolysis via Fe₃O₄ NPs treated enzyme has been employed which produces ～68.0 g/L reducing sugars from sugarcane bagasse. Subsequently, sugar hydrolyzate has been utilized as substrate in the sequential integrated fermentation that produces ～3427.0 mL/L cumulative hydrogen after 408 h. This approach may have potential for the pilot scale production of biohydrogen from waste biomass at low-cost in an eco-friendly manner.     

Impact of low lignin containing brown midrib sorghum mutants to harness biohydrogen production using mixed anaerobic consortia

Three low lignin containing bmr 3 derivatives, namely DRT 07K1, DRT 07K6 and DRT 07K15 developed through backcrossing were used along with the parent, bmr 3 source mutant (IS 21888) for evaluation of biohydrogen production. Results demonstrated that biohydrogen production varied amongst bmr derivatives under similar fermentation conditions. Significant negative correlation was observed between lignin content and fermentative biohydrogen production. All bmr derivatives with lower lignin content produced higher levels of biohydrogen compared to source bmr 3 (IS 21888) which has more lignin content. The maximum and a minimum biohydrogen production observed was 72 and 50 ml/g Total Volatile Solids (TVS) for the DRT 07K6 bmr3 derivative and bmr 3 (IS 21888) respectively. Acetate and butyrate were accounted >85% of volatile fatty acids, indicating acid type fermentations. Statistical analysis revealed that all bmr mutant derivatives with respect to source differ significantly in cumulative biohydrogen production, plant height, grain yield and lignin content. Biohydrogen production from biomass associated at least two different levels, one at lignin entanglement another at the polymeric nature of cellulose and hemicellulose. Further studies are necessary to determine the effect of biomass structure associated with different bmr traits on the microbial growth and biohydrogen production rate.     

Synergistic enhancement of biohydrogen production by supplementing with green synthesized magnetic iron nanoparticles using thermophilic mixed bacteria culture

The production of biohydrogen can be improved by focusing on the nutrients needed by fermentative bacteria like iron. Iron reacts with the [Fe-Fe]-hydrogenase enzyme within the mixed bacteria culture for optimum hydrogen release. Iron nanoparticles (NPs) are attractive due to its unique properties and high reactivity. It can be produced through green synthesis, a more eco-friendly and relatively lower cost process, by using iron salt as precursor and green coconut shell extracted by deep eutectic solvent (DES) as reducing agent. The coconut shell extract consists of phytochemicals that help in producing polydisperse magnetic iron oxide nanoparticles at ～75 nm in size. The addition of optimum concentration of 200 mg Fe/L magnetic iron NPs resulted in the maximum cumulative hydrogen production, glucose utilization and hydrogen yield of 101.33 mL, 9.12 g/L and 0.79 mol H₂/mol glucose respectively. Furthermore, the kinetic analysis on Gompertz model using the optimum magnetic iron NPs concentration showed that the hydrogen production potential (P) and hydrogen production rate (R_m) increased to 50.69 mL and 3.30 mL/h respectively and the lag phase time reduced about 7.12 h as compared with the control experiment (0 mg Fe/L). These results indicated the positive effects of magnetic iron NPs supplementation on fermentative biohydrogen production of mixed bacteria culture and proved the feasibility of adding the magnetic iron NPs as the micronutrient for enhancement of such hydrogen production system.     

Cladophora sp. as a sustainable feedstock for dark fermentative biohydrogen production

Macroalgae are rich in carbohydrates which can be used as a promising substrate for fermentative biohydrogen production. In this study, Cladophora sp. biomass was fermented for biohydrogen production at various inoculum/substrate (I/S) ratios against a control of inoculum without substrate in laboratory-scale batch reactors. The biohydrogen production yield ranged from 40.8 to 54.7 ml H₂/g-VS, with the I/S ratio ranging from 0.0625 to 4. The results indicated that low I/S ratios caused the overloaded accumulation of metabolic products and a significant pH decrease, which negatively affected hydrogen production bacteria's metabolic activity, thus leading to the decrease of hydrogen fermentation efficiency. The overall results demonstrated that Cladophora sp. biomass is an efficient fermentation feedstock for biohydrogen production.     

Multiscale kinetic modeling for biohydrogen production: A study on membrane bioreactors

Hydrogen can be a capable alternative to fossil fuels due to its carbon-free characteristics, in this content, biological hydrogen production is considered a practical approach because technology is green. Due to parameters affecting biohydrogen production, such as operating conditions, it is crucial to predict the process to see the proper yield. There are several conventional and unconventional models used in biohydrogen production prediction. This paper derived a triple first-order prediction model from a previously presented multi-scale kinetic model polynomial built upon the multi-stage growth hypothesis for bio-hydrogen production prediction. The original model was applied to batch and continuous stirred tank reactor studies for their model evaluation, this study evaluated the newly derived model for studies of membrane bioreactors. Due to their increased production yield, membrane bioreactors are an emerging field in biohydrogen production. Although the previous study was mainly applied for batch dark fermentations consisting of various microorganisms, the results presented in this study indicate that it is also applicable for continuous and photo fermentation systems. The original model results reported significant fitness accuracy among different datasheets compared to conventional models like the modified Gompertz model, considering essential factors impacting biohydrogen production suggested in the original model, this paper investigated eleven case studies of dynamic membrane bioreactors with modeling fitness of 99% for most cases. This study reports even higher fitness accuracy compared to the original model, even with different operating conditions.     

Ozone pretreatment of wheat straw for enhanced biohydrogen production

Ozonation was tested as a pretreatment method for enhanced biohydrogen production from wheat straw. Ozone pretreatment effectively degraded wheat straw lignin, and the delignification increased with increase in the applied ozone dose. Results of reducing sugar measurement showed that under our experimental conditions ozone pretreatment significantly increased reducing sugar yields. A simultaneous enzyme hydrolysis and dark fermentation experiment was then conducted using a mixed anaerobic consortium, and the results demonstrated that ozone pretreatment significantly increased biohydrogen production. Compared to the untreated one, hydrogen production in the samples ozonated for 15, 30, 45 and 90 min increased 107%, 134%, 158% and 138%, respectively. Slight inhibitory effect on the dark fermentation was observed with the sample ozonated for 90 min, and the inhibitory effect was due to prolonged ozonation. These results proved that enhancement of biohydrogen production from lignocellulosic biomass using ozone as a pretreatment method is technically feasible.     

Effects of process,operational and environmental variables on biohydrogen production using palm oil mill effluent (POME)

A batch study for biohydrogen production was conducted using raw palm oil mill effluent (POME) and POME sludge as a feed and inoculum respectively. Response Surface Methodology (RSM) was used to design the experiments. Experiments were conducted at different reaction temperatures (30–50 °C), inoculum size to substrate ratios (I:S) and reaction times (8–24 h). An optimum condition of biohydrogen production was achieved with COD removal efficiency of 21.95% with hydrogen yield of 28.47 ml H₂ g^?1 COD removed. The I:S ratio was 40:60, with reaction temperature of 50 °C at 8 h of reaction time. The study showed that a lower substrate concentration (less than 20 g L^?1) for biohydrogen production using pre-settled POME was achievable, with optimum HRT of 8 h under thermophilic condition (50 °C). This study also found that pre-settled POME is feasible to be used as a substrate for biohydrogen production under thermophilic condition.     

Conjoint effect of microwave irradiation and metal nanoparticles on biogas augmentation from anaerobic digestion of green algae

Pretreatment of biomass is a commonly applied technique for improving its biodegradability; however, such methods are energy intensive, which affects the overall efficiency. This study aims to provide an energy efficient solution by combining microwave (MW) pretreatment of algal biomass (Enteromorpha) and metal nanoparticles (NPs). The MW pretreatment of the biomass was in the form of a slurry (liquid:solid 20:1), while pretreatment time and MW pretreatment power were 6min and 600 W, respectively. Nickel (Ni) and Cobalt (Co) NPs with a concentration of 1 mg/L were used. Batch-wise anaerobic digestion was carried out for a period of 264 h. The results showed that MW pretreatment initiates early hydrolysis of green algae thus reducing lag time. NPs had a positive influence in biogas production at the later stages of anaerobic digestion. The highest total biogas production of 53.60ml/gTS was attained by Co NPs + MW pretreatment whereas maximum biohydrogen of 59.52% (v/v) was produced by Ni NPs + MW pretreatment group. Energy analysis showed that combined utilization of MW pretreatment and metal NPs produced added energy while consuming less input energy than MW pretreatment alone. The kinetic parameters were calculated by using modified Gompertz and Logistic function model for each experimental case.     

Production and optimization of high grade cellulase from waste date seeds by Cellulomonas uda NCIM 2353 for biohydrogen production

Production of high grade cellulolytic enzymes from waste agricultural biomass would valorise these wastes to valuable products as well as avoid the pollution problems associated with landfilling of the biomass. In the present study, waste date palm (Phoenix dactylifera) seeds were valorised for cellulase production from Cellulomonas uda NCIM 2353 and for its subsequent usage in biohydrogen production. Optimization of key operational parameters such as date seed concentration, xylose, casein and initial media pH were performed using central composite design to obtain the maximum enzyme yield. The optimum values obtained were (g/L): date seed concentration 30.65, xylose concentration 0.55, casein 7.00 and pH 7.40 for a determination coefficient of 0.999. The results demonstrated a higher prediction accuracy of response surface methodology as the cellulase activity increased six fold (175.96 IU/mL) after optimization. The optimum pH and temperature of purified cellulase was 7 and 50 °C respectively where the enzyme retained nearly 80% of activity upto 180 min. Enzymatic hydrolysis studies showed that a high saccharification efficiency of 60.5% was obtained for acid pretreated sugarcane bagasse by the indigenous cellulase, equivalent to the performance of commercial cellulase. Further, the as-obtained reducing sugars were decomposed by Clostridium thermocellum to produce biohydrogen of maximum concentration 187.44 mmol/L at end of 24 h of fermentation. Results show that date seed substrate based cellulase protein can be employed for industrial processes of biohydrogen production.     

Sulfate- and pH-driven metabolic flexibility in sugarcane vinasse dark fermentation stimulates biohydrogen evolution,sulfidogenesis or homoacetogenesis

Dark fermentation of sugarcane vinasse can be used as a “cleaning” step to remove sulfate prior to methanogenesis because sulfidogenic conditions can be successfully established in parallel with biohydrogen production. Using a 2² central composite rotational design (CCRD) and response surface methodology (RSM), this study assessed the impacts of bicarbonate and sulfate availability on the establishment of sulfidogenesis in the thermophilic (55 °C) fermentation of vinasse in batch reactors, equally assessing the impacts on biohydrogen evolution. CCRD-RSM results indicated the favoring of biohydrogen production at the lowest sulfate and bicarbonate concentrations, whilst the opposite was observed for sulfidogenesis. Glycerol, lactate, and hydrogen were the preferential electron donors utilized by sulfate-reducing bacteria (SRB), whilst ethanol was markedly consumed only at high sulfate concentrations. SRB were inhibited by sodium when dosing excess NaHCO₃ and Na₂SO₄. Complementary tests revealed maximum biohydrogen production (2.40 mmol) out of the CCRD, at pH exceeding 7.5 with no interference of sulfidogenesis. Non-efficient biohydrogen production was observed at low pH (<5.0; ～1.90 mmol) because the uptake of lactate was inhibited. Meanwhile, homoacetogenesis was established under intermediate pH range (5.5–6.5), as revealed by the accumulation of acetate (up to 2.5 g L^?1). 16S rRNA gene amplicon sequencing further revealed the genera Thermoanaerobacterium/Pseudoclostridium, Desulfotomaculum/Desulfohalotomaculum and Sporomusaceae/Moorella as the main biohydrogen-producing, sulfate-removing and biohydrogen-consuming (homoacetogens) microbial groups, respectively. Hence, using a single inoculum source, vinasse may provide a butyrate-rich (along with biohydrogen-rich biogas) or a sulfate-free and acetate-rich fermented effluent, depending mainly on proper pH control.     

Biohydrogen from food waste: Modeling and estimation by machine learning based super learner approach

This study demonstrated the application of a hybrid Bayesian algorithm (BA) and support vector regression (SVR) as a potential super-learner tool (BA-SVR) to predict biohydrogen production from food waste-originated feedstocks. The novelty of the present approach, as compared to the existing response surface methodology (RSM), includes (i) hybridization of BA with SVR for modeling of biohydrogen production and minimization of biomethane formation, (ii) performance evaluation and comparison of the developed BA-SVR models with the existing RSM models based on the several indicators such as coefficient of determination (R²), relative error (RE), mean absolute error (MAE), mean absolute percentage error (MAPE), and root mean square error (RMSE), (iii) analysis of the robustness of the model and (iv) testing generalization ability. The calculated values of these indicators suggested that the proposed super leaner models demonstrated better performance predicting the biohydrogen and biomethane (products) responses than those using the existing RSM models - as reported in Rafieenia et al. 2019 [45]. The estimated low errors for biohydrogen: MAE = 0.5919, RMSE = 0.592, MAPE = 11.1387; for biomethane: MAE = 0.2681, RMSE = 0.2688, MAPE = 0.3708, signifie the reliable model predictions. The BA-SVR model also provided high adj R² (>0.99 for both biohydrogen and biomethane), indicating an excellent fitting of the model. Concerning the MAPE, the proposed BA-SVR models for both the biohydrogen and biomethane responses showed superior performances (as compared to the RSM models) with a performance enhancement of 64.16% and 98.81%, respectively.     

Effect of silica-core gold-shell nanoparticles on the kinetics of biohydrogen production and pollutant hydrogenation via organic acid photofermentation over enhanced near-infrared illumination

A biological photoinduced fermentation process provides an alternative to traditional hydrogen productions. In this study, biohydrogen production was investigated at near IR region coupled to a near-field enhancement by silica-core gold-shell nanoparticles (NPs) over a range of acetate concentrations (5–40 mM) and light intensities (11–160 W/m²). The kinetic data were modeled using modified Monod equations containing light intensity effects. The yields of H₂ and CO₂ produced per acetate were determined as 2.31 mol-H₂/mol-Ac and 0.83 mol-CO₂/mol-Ac and increased to 4.38 mmol-H₂/mmol-Ma and 2.62 mmol-CO₂/mmol-Ma when malate was used. Maximum increases in H₂ and CO₂ productions by 115% and 113% were observed by adding NPs without affecting the bacterial growth rates (6.1–8.2 mg-DCM/L/hour) while the highest hydrogen production rate was determined as 0.81 mmol/L/hour. Model simulations showed that the energy conversion efficiency increased with NPs concentration but decreased with the intensity. Complete hydrogenation application was demonstrated with toxic 2-chlorobiphenyl using Pd catalysts.