Optimization of decolorization of methylene blue (MB) dye by lignin peroxidase (LiP) enzyme produced by white-rot fungus Phanerochaete chrysosporium using sewage treatment plant (STP) sludge as a major substrate was carried out in the laboratory. Optimization by the one-factor-at-a-time (OFAT) and statistical approach was carried out to determine the process conditions on optimum decolorization of MB dye using LiP enzyme in static mode. The OFAT method indicated that the optimum conditions for decolorization of MB dye (removal: 14-40%) was at temperature 55 degrees C, pH 5.0 with hydrogen peroxide (H(2)O(2)) concentration 4.0mM, MB dye concentration 20mg/L and LiP activity 0.487U/ml. The addition of veratryl alcohol to the reaction mixtures did not contribute any further increases in decolorization. The initial concentration of MB and the activity of LiP enzyme were further optimized using response surface methodology (RSM). The contour and surface plots suggested that the optimum initial concentration of MB and LiP activity predicted were 15mg/L and 0.687U/ml, respectively for the removal of 65%. The validation of the model showed that the decolorization process gave the higher removal of 90% in agitation mode compared to the static mode with 65% for 60min of incubation time by LiP enzyme.
Biogas is an economical and environmentally friendly renewable energy which can be produced by anaerobic digestion (AD). This biochemical method converts organic compounds (mainly from wastes) into a sustainable source of energy. Anaerobic co-digestion (AcoD) is a method combining more than one substrate to resolve the difficulties faced in a single substrate AD system. Solid wastes increases as the population increase so do the urbanization and industrial industries. Food waste and sewage sludge are examples of one of the solid wastes. Co-digesting of both substrates may improve process stabilization to increase biogas production and overcome the nutrients imbalance. Thus, anaerobic co-digestion has been recognized as a technology that could provide a clean renewable energy source and helps reduce the landfill problem. The objective of this paper is to investigate the recent achievements and perspectives on the interaction of co-digestion between food waste and sewage sludge to improve biogas production. This may provide valuable information on the optimization of combinations of substrates: food waste and sewage sludge and prediction of bioreactor performance.
Recently, the production of renewable biogas such as biohydrogen and biomethane from wastewaters through anaerobic fermentation has gained worldwide attention. In the present study, a mobile bioenergy generation station had been constructed based on a high-efficiency hydrogenesis & methanogenesis technology (HyMeTek) developed by Feng Chia University, Taiwan. The substrate was a beverage wastewater having chemical oxygen demand (COD) concentration of 1200 mg/L. This bioenergy station had a feedstock tank (3.8 m3), a nutrient tank (0.8 m3), an acidogenesis tank (AT, 2 m3), two methanogenesis tanks (MT, 4 m3 for each), a membrane bioreactor and a control room. Biogas production rate, methane concentration, COD removal efficiencies, energy efficiency and economical interest of the plant were assessed. The peak total methane production rates for AT (at hydraulic retention time, HRT, 4 h) and MT (at HRT 8 h) were 430 and 7 mL/L·d, respectively. A strategy of shortening HRT was a promising method to enhance biogas quality and energy efficiency. This mobile bioenergy system has commercial potential because it could bring good economic benefit of initial rate of return (58.84%) and payback time (2.68 y).
Mixed culture sludge has been widely used as a microbial consortium for biohydrogen production. Simple thermal treatment of sludge is usually required in order to eliminate any H2-consuming bacteria that would reduce H2 production. In this study, thermal treatment of sludge was carried out at various temperatures. Electron flow model was then applied in order to assess community structure in the sludge upon thermal treatment for biohydrogen production. Results show that the dominant electron sink was acetate (150-217 e- meq/mol glucose). The electron equivalent (e- eq) balances were within 0.8-18% for all experiments. Treatment at 100 °C attained the highest H2 yield of 3.44 mol H2/mol glucose from the stoichiometric reaction. As the treatment temperature increased from 80 to 100 °C, the computed acetyl-CoA and reduced form of ferredoxin (Fdred) concentrations increased from 13.01 to 17.34 e- eq (1.63-2.17 mol) and 1.34 to 4.18 e- eq (0.67-2.09 mol), respectively. The NADH2 balance error varied from 3 to 10% and the term e-(Fd↔NADH2) (m) in the NADH2 balance was NADH2 consumption (m = -1). The H2 production was mainly via the Fd:hydrogenase system and this is supported with a good NADH2 balance. Using the modified Gompertz model, the highest maximum H2 production potential was 1194 mL whereas the maximum rate of H2 production was 357 mL/h recorded at 100 °C of treatment.
The application of the anammox process has great potential in treating nitrogen-rich wastewater. The presence of Fe (II) is expected to affect the growth and activity of anammox bacteria. Short-term (acute) and long-term effects (chronic) of Fe (II) on anammox activity were investigated. In the short-term study, results demonstrated that the optimum concentration of Fe (II) that could be added to anammox is 0.08 mM, at which specific anammox activity (SAA) improved by 60% compared to the control assay, 0.00 mM. The inhibition concentration, IC50, of Fe (II) was found to be 0.192 mM. Kinetics of anammox specific growth rate were estimated based on results of the batch test and evaluated with Han-Levenspiel's substrate inhibition kinetics model. The optimum concentration and IC50 of Fe (II) predicted by the Han-Levenspiel model was similar to the batch test, with values of 0.07 mM and 0.20 mM, respectively. The long-term effect of Fe (II) on the performance of a sequencing batch reactor (SBR) was evaluated. Results showed that an appropriate Fe (II) addition enhanced anammox activity, achieving 85% NH4+-N and 96% NO2--N removal efficiency when 0.08 mM of Fe (II) was added. Quantitative polymerase chain reaction (qPCR) was adopted to detect and identify the anammox bacteria.
Bacteriocin-like inhibitory substances (BLIS) produced by Lactococcus lactis Gh1 had shown antimicrobial activity against Listeria monocytogenes ATCC 15313. Brain Heart Infusion (BHI) broth is used for the cultivation and enumeration of lactic acid bacteria, but there is a need to improve the current medium composition for enhancement of BLIS production, and one of the approaches is to model the optimization process and identify the most appropriate medium formulation. Response surface methodology (RSM) and artificial neural network (ANN) models were employed in this study. In medium optimization, ANN (R2 = 0.98) methodology provided better estimation point and data fitting as compared to RSM (R2 = 0.79). In ANN, the optimal medium consisted of 35.38 g/L soytone, 16 g/L fructose, 3.25 g/L sodium chloride (NaCl) and 5.40 g/L disodium phosphate (Na2HPO4). BLIS production in optimal medium (717.13 ± 0.76 AU/mL) was about 1.40-fold higher than that obtained in nonoptimised (520.56 ± 3.37 AU/mL) medium. BLIS production was further improved by about 1.18 times higher in 2 L stirred tank bioreactor (787.40 ± 1.30 AU/mL) as compared to that obtained in 250 mL shake flask (665.28 ± 14.22 AU/mL) using the optimised medium.
Waste management in Malaysia remains a persistent economic and environmental challenge. Up to date, more than 80% of Malaysian solid waste disposed at landfills and dumpsites. Therefore, Malaysia is facing an urgent need to move towards a sustainable solid waste management and thus resource recovery from organic solid waste. Hence, this study aims to investigate the feasibility of energy and bio fertilizer recovery from organic fraction municipal solid waste (OFMSW) via anaerobic digestion. The economic and environmental benefit analysis was investigated. Approximate and elementary analysis of OFMSW samples were carried out to estimate the potential production of biogas and bio fertilizer. It was found that organic waste contributes about 45% of the total MSW generated in Malaysia. Anaerobic digestion of 50% of organic waste is expected to produce 3941 MWh/day of electrical energy and 2500 t/day of bio fertilizer. In terms of environmental impacts, 2735 t/day of Carbon dioxide (CO2) emission, 1128 m2/day of landfilling area and 481 m3/day of leachate can be avoided. A net revenue of 3300 million RM (1 US Dollar ≈ 4.15 RM) can be generated by the sales of electricity via Feed-in-Tariff (FiT), sales of biofertilizer to local agricultural industries and inclusive of the saving generated from the reduction of OFMSW landfilling operations and leachate treatment at landfills. Economic development can go hand-in-hand with environmental sound practices in the field of waste management.
In this study, a selected γ-aminobutyric acid (GABA)-rich Malaysian strain Aspergillus oryzae NSK was collected from soy sauce koji. The strain was used to explore the effect of using renewable native sugar syrup, sugarcane, nipa, and molasses as fermentable substrates for developing a novel functional GABA soy sauce. We evaluated the strain using the chosen native sugars for 7 days using shake flask fermentation at 30 °C. The results showed optimum GABA concentration was achieved using cane molasses as the fermentable substrate (354.08 mg/L), followed by sugarcane syrup (320.7 mg/L) and nipa syrup (232.07 mg/L). Cane molasses was subsequently utilized as a substrate to determine the most suitable concentration for A. oryzae NSK to enhance GABA production and was determined as 50% g/L of glucose standard cane molasses. Our findings indicate that cane molasses can be used as a GABA-rich ingredient to develop a new starter culture for A. oryzae NSK soy sauce production.
The in vitro shoot proliferation of endemic Begonia pavonina in three culture conditions i.e semisolid medium (SM), liquid culture medium (LM) and in temporary immersion bioreactor system (RITA®) was analyzed in this study. To minimize contamination rates, seeds were surface sterilized and cultured on MS basal media. The clean raised shoots were then used as explants for inoculation onto the tested culture conditions. In this experiment, the explants were maintained in MS medium supplemented with 0.1mgL-1 BAP for shoot multiplication. After 4 weeks of incubation, higher regeneration rates were observed in TIM as compared to other medium conditions. The maximum shoot number was obtained from TIM system with a mean of 5.30 shoots per explant, followed by LM (2.47 shoots per explant) and SM (1.2 shoots per explant). Shoot hyperhydration was also lowest in a TIM system. Overall, TIM was shown to produce higher shoot multiplications combined with healthy morphological characteristics of plantlets. Shoot cultures from the all cultures were successfully rooted in vitro and acclimatized well in the greenhouse.
For the anaerobic biological treatment of saline wastewater, Anaerobic Digestion (AD) is currently a possibility, even though elevated salt concentrations can be a major obstacle. Anaerobic consortia and especially methanogenic archaea are very sensitive to fluctuations in salinity. When working with Upflow Sludge Blanket Reactor (UASB) technology, in which the microorganisms are aggregated and retained in the system as a granular biofilm, high sodium concentration negatively affects aggregation and consequently process performances. In this research, we analysed the structure of the biofilm and granules formed during the anaerobic treatment of high salinity (at 10 and 20 g/L of sodium) synthetic wastewater at lab scale. The acclimated inoculum was able to accomplish high rates of organics removal at all the salinity levels tested. 16S rRNA gene clonal analysis and Fluorescence In Situ Hybridization (FISH) analyses identified the acetoclastic Methanosaeta harundinacea as the key player involved acetate degradation and microbial attachment/granulation. When additional calcium (1 g/L) was added to overcome the negative effect of sodium on microbial aggregation, during the biofilm formation process microbial attachment and acetate degradation decreased. The same result was observed on granules formation: while calcium had a positive effect on granules strength when added to UASB reactors, Methanosaeta filaments were not present and the degradation of the partially acidified substrate was negatively influenced. This research demonstrated the possibility to get granulation at high salinity, bringing to the forefront the importance of a selection towards Methanosaeta cells growing in filamentous form to obtain strong and healthy granules.
Optimization of fermentation processes requires monitoring the species composition of starter cultures and their growth during fermentation. Most starter cultures contain closely related species. Nowadays, high-resolution melting (HRM) analysis is extensively used for multiplex identification of closely related species. In the present paper, we applied real-time polymerase chain reaction (PCR) with HRM analysis for the detection and differentiation of Lactobacillus sakei and L. curvatus. A primer pair was selected for the site of the rpoA gene of Lactobacillus spp. Eleven starter cultures and fifteen fermented sausages with a known bacterial composition were successfully tested using real-time PCR with HRM analysis with the developed primer pair.
Pseudomonas sp. LAB-08 was isolated from a phenol-fed bioreactor constructed with contaminated aquifer soil as the inoculum. Strain LAB-08 utilized phenol as a sole carbon and energy source. Here, we report the genome sequence and annotation of Pseudomonas sp. LAB-08.
The extensive amount of available information on global warming suggests that this issue has become prevalent worldwide. Majority of countries have issued laws and policies in response to this concern by requiring their industrial sectors to reduce greenhouse gas emissions, such as CO2. Thus, introducing new and more effective treatment methods, such as biological techniques, is crucial to control the emission of greenhouse gases. Many studies have demonstrated CO2 fixation using photo-bioreactors and raceway ponds, but a comprehensive review is yet to be published on biological CO2 fixation. A comprehensive review of CO2 fixation through biological process is presented in this paper as biological processes are ideal to control both organic and inorganic pollutants. This process can also cover the classification of methods, functional mechanisms, designs, and their operational parameters, which are crucial for efficient CO2 fixation. This review also suggests the bio-trickling filter process as an appropriate approach in CO2 fixation to assist in creating a pollution-free environment. Finally, this paper introduces optimum designs, growth rate models, and CO2 fixation of microalgae, functions, and operations in biological CO2 fixation.
The production of ethanol, from glucose in batch and fed batch culture, was investigated. In the fed batch culture, the glucose feeding was added into the culture at 16th hour of fermentation. The effects of different glucose concentration feeding rates on ethanol fermentation were investigated for fed batch culture. The 2gL-1hr-1 glucose concentration feeding rate was found to give higher ethanol yield (2.47 g ethanol g glucose-1), with respect to substrate consumed as compared to 8 gL-1hr-1 (0.23 g ethanol g glucose-1) and 4 gL-1hr-1 (0.20 g ethanol g glucose-1). The ethanol yield with respect to substrate consumed obtained in batch culture was 0.81 g ethanol g glucose-1. The fed batch culture at 2 gL-1hr-1 glucose concentration feeding rate was proven to be a better fermentation system than the batch culture. The specific growth rate, specific glucose consumption rate and specific ethanol production rate for the fed batch fermentation, at 2 gL-1hr-1 glucose concentration feeding rate, were 0.065 hr-1, 1.20 hr-1 and 0.0009 hr-1, respectively.
Water-immiscible substrate, diesel, was supplied as the main substrate in the fermentation of Pseudomonas aeruginosa USM-AR2 producing rhamnolipid biosurfactant, in a stirred tank bioreactor. In addition to the typical gas-aqueous system, this system includes gas-hydrocarbon-aqueous phases and the presence of surfactant (rhamnolipid) in the fermentation broth. The effect of diesel dispersion on volumetric oxygen transfer coefficient, k L a, and thus oxygen transfer, was evaluated at different agitations of 400, 500 and 600 rpm. The oxygen transfer in this oil-water-surfactant system was shown to be affected by different oil dispersion at those agitation rates. The highest diesel dispersion was obtained at 500 rpm or impeller tip speed of 1.31 m/s, compared to 400 and 600 rpm, which led to the highest k L a, growth and rhamnolipid production by P. aeruginosa USM-AR2. This showed the highest substrate mixing and homogenization at this agitation speed that led to the efficient substrate utilization by the cells. The oxygen uptake rate of P. aeruginosa USM-AR2 was 5.55 mmol/L/h, which showed that even the lowest k L a (48.21 h-1) and hence OTR (57.71 mmol/L/h) obtained at 400 rpm was sufficient to fulfill the oxygen demand of the cells. The effect of rhamnolipid concentration on k L a showed that k L a increased as rhamnolipid concentration increased to 0.6 g/L before reaching a plateau. This trend was similar for all agitation rates of 400, 500 and 600 rpm, which might be due to the increase in the resistance to oxygen transfer (k L decrease) and the increase in the specific interfacial area (a).
Oleochemicals industry effluence mainly contains a high chemical oxygen demand (COD) in a range of 6000-20,000 ppm. An effective biological wastewater treatment process must be carried out before wastewater is discharged into the environment. In this study, a submerged bed biofilm reactor (SBBR) was adapted to the biological oleochemical wastewater treatment plant observed in the present study. The effect of wastewater flow rate (100-300 mL/min), Cosmoball® percentage in the SBBR system (25-75%), and percentage of activated sludge (0-50%) were investigated in terms of COD reduction. The Box-Behnken design was used for response surface methodology (RSM) and to create a set of 18 experimental runs, which was needed for optimising the biological oleochemical wastewater treatment. A quadratic polynomial model with estimated coefficients was developed to describe COD reduction patterns. The analysis of variance (ANOVA) shows that the wastewater flow rate was the most effective factor in reducing COD, followed by activated sludge percentage and Cosmoball® carrier percentage. Under the optimum conditions (i.e., a wastewater flow rate of 103.25 mL/min a Cosmoball® carrier percentage of 71.94%, and an activated sludge percentage of 40.50%) a COD reduction of 98% was achieved. Thus, under optimum conditions, as suggested by the BBD, SBBR systems can be used as a viable means of biological wastewater treatment in the oleochemicals industry.
The present study intends to evaluate the potential of co-digestion for utilizing Organic fraction of Municipal Solid Waste (OFMSW) and sewage sludge (SS) for enhanced biogas production. Metagenomic analysis was performed to identify the dominant bacteria, archaea and fungi, changes in their communities with time and their functional roles during the course of anaerobic digestion (AD). The cumulative biogas yield of 586.2 mL biogas/gVS with the highest methane concentration of 69.5% was observed under an optimum ratio of OFMSW:SS (40:60 w/w). Bacteria and fungi were found to be majorly involved in hydrolysis and initial stages of AD. Probably, the most common archaea Methanosarsina sp. primarily followed the acetoclastic pathway. The hydrogenotrophic pathway was less followed as indicated by the reduction in abundance of syntrophic acetate oxidizers. An adequate understanding of microbial communities is important to manipulate and inoculate the specific microbial consortia to maximize CH4 production through AD.
The novel immobilized microbial granules (IMG) shows a significant effect of nitrification for freshwater aquaculture. However, there is lack of evaluation study on the performance of nitrification at high salinity due to the concentration of recycled water or seawater utilization. A laboratory scale moving bed bioreactor (MBBR) with IMG was tested on recycled synthetic aquaculture wastewater for the nitrification at 2.5 mg/L NH3-N daily. The results indicated that IMG showed a high salinity tolerance and effectively converted ammonia to nitrate up to 92% at high salinity of 35.0 g/L NaCl. As salinity increased from near zero to 35.0 g/L, the microbial activity of nitrite oxidation bacteria (NOB) in the IMG decreased by 86.32%. The microbial community analysis indicated that salinity significantly influenced the community structure. It was found that Nitrosomonas sp. and Nitrospira sp. were the dominant genera for ammonia oxidation bacteria (AOB) and NOB respectively at different salinity levels.
Anammox bacteria can easily undergo starvation due to fluctuations in feed flowrate and concentration in wastewater treatment plants. In this study, we analyzed the effects of different types of storage conditions (presence of ammonium (Ra), nitrite (Rn), hydrazine (Rh), and no substrate (Rc)) in aiding the viability of anammox bacteria during starvation and recovery. After starvation, the bacteria were subjected to a 15-week recovery period. Anammox bacteria showed better results during starvation and recovery in Rh as compared to other conditions. Decay rate values obtained after starvation in Ra, Rn, Rh, and Rc were 0.032/day, 0.042/day, 0.019/day, and 0.037/day, respectively. Meanwhile, µmax values obtained in Rh, Ra, Rn, and Rc on the 15th week of recovery were 0.092, 0.075, 0.011, and 0.067 d-1, respectively. This indicated that the availability of hydrazine helps to reduce the mortality rate of anammox bacteria during starvation and enhances the recovery of anammox process.
Acetone-butanol-ethanol (ABE) fermentation from Palm Oil Mill Effluent (POME) by C. acetobutylicum NCIMB 13357 in an oscillatory flow bioreactor was investigated. Experimental works were conducted in a U-shaped stainless steel oscillatory flow bioreactor at oscillation frequency between 0.45-0.78 Hz and a constant amplitude of 12.5 mm. Fermentations were carried out for 72 hr at 35oC using palm oil mill effluent and reinforced clostridia medium as a growth medium in batch culture. Result of this investigation showed that POME is a viable media for ABE fermentation and oscillatory flow bioreactor has an excellent potential as an alternative fermentation device.