One-time ultrasonication pre-treatment of Rhodobacter sphaeroides was evaluated for improving biohydrogen production via photofermentation. Batch experiments were performed by varying ultrasonication amplitude (15, 30, and 45%) and duration (5, 10, and 15 min) using combined effluents from palm oil as well as pulp and paper mill as a single substrate. Experimental data showed that ultrasonication at amplitude 30% for 10 min (256.33 J/mL) achieved the highest biohydrogen yield of 9.982 mL H2/mLmedium with 5.125% of light efficiency. A maximum CODtotal removal of 44.7% was also obtained. However, when higher ultrasonication energy inputs (>256.33 J/mL) were transmitted to the cells, biohydrogen production did not improve further. In fact, 20.6% decrease of biohydrogen yield (as compared to the highest biohydrogen yield) was observed using the most intense ultrasonicated inoculum (472.59 J/mL). Field emission scanning electron microscope images revealed the occurrence of cell damages and biomass losses if ultrasonication at 472.59 J/mL was used. The present results suggested that moderate ultrasonication pre-treatment was an effective technique to improve biohydrogen production performances of R. sphaeroides.
Although a significant amount of brewery wastewater (BW) is generated during beer production, the nutrients in the BW could be reused as a potential bio-resource for biohydrogen production. Therefore, improvements in photofermentative biohydrogen production due to a combination of BW and pulp and paper mill effluent (PPME) as a mixed production medium were investigated comprehensively in this study. The experimental results showed that both the biohydrogen yield and the chemical oxygen demand removal were improved through the combination of BW and PPME. The best biohydrogen yield of 0.69 mol H2/L medium was obtained using the combination of 10 % BW + 90 % PPME (10B90P), while the reuse of the wastewater alone (100 % BW and 100 % PPME) resulted in 42.3 and 44.0 % less biohydrogen yields than the highest yield, respectively. The greatest light efficiency was 1.97 % and was also achieved using the combination of both wastewaters at 10B90P. This study revealed the potential of reusing and combining two different effluents together, in which the combination of BW and PPME improved the nutrients and light penetration into the mixed production medium.
The enhancement of lignocellulose hydrolysis using enzyme complexes requires an efficient pretreatment process to obtain susceptible conditions for the enzyme attack. This study focuses on removing a major part of the lignin layer from kenaf (Hibiscus cannabinus) while simultaneously maintaining most of the hemicellulose. A two-stage pretreatment process is adopted using calcium hydroxide, Ca(OH)₂, and peracetic acid, PPA, to break the recalcitrant lignin layer from other structural polysaccharides. An experimental screening of several pretreatment chemicals, concentrations, temperatures and solid-liquid ratios enabled the production of an optimally designed pretreatment process for kenaf. Our results showed that the pretreatment process has provide 59.25% lignin removal while maintaining 87.72% and 96.17% hemicellulose and cellulose, respectively, using 1g of Ca(OH)₂/L and a 8:1 (mL:g) ratio of liquid-Ca(OH)₂ at 50 °C for 1.5 h followed by 20% peracetic acid pretreatment at 75 °C for 2 h. These results validate this mild approach for aiding future enzymatic hydrolysis.
It is widely believed that reducing the lignocellulosic biomass particle size would improve the biomass digestibility by increasing the total surface area and eliminating mass and heat transfer limitation during hydrolysis reactions. However, past studies demonstrate that particle size influences biomass digestibility to a limited extent. Thus, this paper studies the effect of particle size (milled: 2 mm, 5 mm, cut: 2 cm and 5 cm) on rice straw conversion. Two different Ammonia Fiber Expansion (AFEX) pretreament conditions, AFEX C1 (low severity) and AFEX C2 (high severity) are used to pretreat the rice straw (named as AC1RS and AC2RS substrates respectively) at different particle size.
E. coli has been engineered to produce xylitol, but the production faces bottlenecks in terms of production yield and cell viability. In this study, recombinant E. coli (rE. coli) was immobilized on untreated and treated multiwalled carbon nanotubes (MWCNTs) for xylitol production. The immobilized rE. coli on untreated MWCNTs gave the highest xylitol production (5.47 g L-1) and a productivity of 0.22 g L-1 h-1. The doubling time for the immobilized cells increased up to 20.40 h and was higher than that of free cells (3.67 h). Cell lysis of the immobilized cells was reduced by up to 73 %, and plasmid stability improved by up to 17 % compared to those of free cells. Xylitol production using the optimum parameters (pH 7.4, 0.005 mM and 29 °C) achieved a xylitol production and productivity of 6.33 g L-1 and 0.26 g L-1 h-1, respectively. A seven-cycle repeated batch fermentation was carried out for up to 168 h, which showed maximum xylitol production of 7.36 g L-1 during the third cycle. Hence, this new adsorption immobilization system using MWCNTs is an alternative to improve the production of xylitol.
Adsorption technology has led to the development of promising techniques to purify biogas, i.e., biomethane or biohydrogen. Such techniques mainly depend on the adsorbent ability and operating parameters. This research focused on adsorption technology for upgrading biogas technique by developing a novel adsorbent. The commercial coconut shell activated carbon (CAC) and two types of gases (H2S/N2 and H2S/N2/CO2) were used. CAC was modified by copper sulfate (CuSO4), zinc acetate (ZnAc2), potassium hydroxide (KOH), potassium iodide (KI), and sodium carbonate (Na2CO3) on their surface to increase the selectivity of H2S removal. Commercial H2S adsorbents were soaked in 7 wt.% of impregnated solution for 30 min before drying at 120°C for 24 h. The synthesized adsorbent's physical and chemical properties, including surface morphology, porosity, and structures, were characterized by SEM-EDX, FTIR, XRD, TGA, and BET analyses. For real applications, the modified adsorbents were used in a real-time 0.85 L single-column adsorber unit. The operating parameters for the H2S adsorption in the adsorber unit varied in L/D ratio (0.5-2.5) and feed flow rate (1.5-5.5 L/min) where, also equivalent with a gas hourly space velocity, GHSV (212.4-780.0 hour-1) used. The performances of H2S adsorption were then compared with those of the best adsorbent that can be used for further investigation. Characterization results revealed that the impregnated solution homogeneously covered the adsorbent surface, morphology, and properties (i.e., crystallinity and surface area). BET analysis further shows that the modified adsorbents surface area decreased by up to 96%. Hence, ZnAc2-CAC clarify as the best adsorption capacity ranging within 1.3-1.7 mg H2S/g, whereby the studied extended to adsorption-desorption cycle.
A two-stage fermentation process consisting of dark and photo-fermentation periods was carried out in a batch reactor. In the first stage, glucose was fermented in the dark stage using Clostridium saccharoperbutylacetonicum N1-4 (ATCC 13564; CSN1-4) to produce acetate, CO2 and H2. The acetate produced in the first stage is fermented to H2 and CO2 by Rhodobacter sphaeroides NCIMB 8253 for further hydrogen production in the second, illuminated stage. The yield of hydrogen in the first stage was about 3.10 mol H2 (mol glucose)(-1) at a glucose concentration of 10 g L(-1), pH 6 +/- 0.2 and 37 degrees C and the second stage yield was about 1.10-1.25 mol H2 (mol acetic acid)(-1) at pH 6.8 +/- 0.2 and 32 degrees C, without removal of the Clostridium CSN1-4. The overall yield of hydrogen in the two-stage process, with glucose as the main substrate was higher than single-stage fermentation.
Box-Wilson design (BWD) model was applied to determine the optimum values of influencing parameters in anaerobic fermentation to produce hydrogen using Clostridium saccharoperbutylacetonicum N1-4 (ATCC 13564). The main focus of the study was to find the optimal relationship between the hydrogen yield and three variables including initial substrate concentration, initial medium pH and reaction temperature. Microbial growth kinetic parameters for hydrogen production under anaerobic conditions were determined using the Monod model with incorporation of a substrate inhibition term. The values of micro(max) (maximum specific growth rate) and K, (saturation constant) were 0.398 h(-1) and 5.509 g L(-1), respectively, using glucose as the substrate. The experimental substrate and biomass-concentration profiles were in good agreement with those obtained by the kinetic-model predictions. By varying the conditions of the initial substrate concentration (1-40 g L(-1)), reaction temperature (25-40 degrees C) and initial medium pH (4-8), the model predicted a maximum hydrogen yield of 3.24 mol H2 (mol glucose)(-1). The experimental data collected utilising this design was successfully fitted to a second-order polynomial model. An optimum operating condition of 10 g L(-1) initial substrate concentration, 37 degrees C reaction temperature and 6.0 +/- 0.2 initial medium pH gave 80% of the predicted maximum yield of hydrogen where as the experimental yield obtained in this study was 77.75% exhibiting a close accuracy between estimated and experimental values. This is the first report to predict bio-hydrogen yield by applying Box-Wilson Design in anaerobic fermentation while optimizing the effects of environmental factors prevailing there by investigating the effects of environmental factors.
A Pichia pastoris transformant carrying the cutinase cDNA of Glomerella cingulata was over-expressed in a 5L bioreactor (2.0L working volume) under fed-batch conditions. Bioreactor experiments rely on varying selected parameters in repeated rounds of optimisation: here these included duration of induction, pH and temperature. Highest cell densities (320gL(-1) wet cell weight) with a cutinase production of 3800mgL(-1) and an activity of 434UmL(-1) were achieved 24h after induction with methanol in basal salt medium (at pH 5 and 28°C). Characterisation of the cutinase showed that it was stable between pH 6 and pH 11, had an optimum pH of 8.0 and retained activity for 30min at 50°C (optimum temperature 25°C).The preferred substrates of G. cingulata cutinase were the medium- to long-chain ρ-nitrophenyl esters of ρ-nitrophenylcaprylate (C8), ρ-nitrophenyllaurate (C12) and ρ-nitrophenylmyristate (C14), with the highest catalytic efficiency, kcat/Km of 7.7±0.7mM(-1)s(-1) for ρ-nitrophenylcaprylate. Microscopic analyses showed that the G. cingulata cutinase was also capable of depolymerising the high molecular weight synthetic polyester, polyethylene terephthalate.
The performances of various anhydrous and aqueous choline chloride-dicarboxylic acid based deep eutectic solvents (DESs) were evaluated for furfural production from oil palm fronds without any additional catalyst. The effects of different carbon chain length dicarboxylic acids and water content in each DES on furfural production were investigated. Oil palm fronds, DES and water (0-5 ml) were mixed and reacted in an oil bath (60-300 min). Reacted oil palm fronds had the potential to be reused as cellulose-rich-valuable by-products. At 100 °C, aqueous choline chloride-oxalic acid (16.4 wt% H2O) produced the highest furfural yield of 26.34% and cellulose composition up to 72.79% in the reacted oil palm fronds. Despite operating at suitable reaction duration for dicarboxylic acid with longer carbon chain length, aqueous choline chloride-malonic acid and aqueous choline chloride-succinic acid performed poorly with furfural yield of less than 1%.
Deep eutectic solvents (DESs) have received considerable attention in recent years due to their low cost, low toxicity, and biodegradable properties. In this study, a sequential pretreatment comprising of a DES (choline chloride:urea in a ratio of 1:2) and divalent inorganic salt (CuCl2) was evaluated, with the aim of recovering xylose from oil palm fronds (OPF). At a solid-to-liquid ratio of 1:10 (w/v), DES alone was ineffective in promoting xylose extraction from OPF. However, a combination of DES (120°C, 4h) and 0.4mol/L of CuCl2 (120°C, 30min) resulted in a pretreatment hydrolysate containing 14.76g/L of xylose, remarkably yielding 25% more xylose than the CuCl2-only pretreatment (11.87g/L). Characterization studies such as FE-SEM, BET, XRD, and FTIR confirmed the delignification of OPF when DES was implemented. Thus, the use of this integrated pretreatment system enabled xylose recoveries which were comparable with other traditional pretreatments.