Packed-bed Bioreactor Research Articles

Increasing the acetification rate of Acetobacter aceti adsorbed on luffa sponge using recycle of incremental oxygenated medium.

Speeding up the production of vinegar from rice wine by acetification, using a packed-bed bioreactor with a luffa sponge matrix (LSM) as adsorption carrier of acetic acid bacteria (AAB), and the effect of oxygenation of the recycled medium were investigated. The 0.06 L/min recycle of medium resulted in a high oxygen-transfer coefficient, while optimal dissolved oxygen (DO) of the medium maximized planktonic AAB cell growth with no contamination due to high acid in an external reservoir without LSM. The highest acetification rate (ETA) of 2.857 ± 0.1g/L/day was achieved with DO 3.5-4.5ppm at 35 ± 1°C. To increase ETA, the optimized oxygenated medium was externally supplied and recycled at the ratio of 0.1. Therefore, acetification was conducted in both the bioreactor and reservoir resulting in an increased ETA (6 ± 0.2g/L/day). This also aligned with the highest system AAB biomass (confirmed by scanning electron microscopy). Under the recycled oxygenated medium supply consistently high biotransformation yields (average 77.3%) were observed over nine sequential cycles. Meanwhile, an average ETA of 6.3 ± 0.2g/L/day was obtained. This method can have practical applications in improving the efficiency and speeding up small-scale vinegar production.

β-Mannanase Production Using Coffee Industry Waste for Application in Soluble Coffee Processing.

Soluble coffee offers the combined benefits of high added value and practicality for its consumers. The hydrolysis of coffee polysaccharides by the biochemical route, using enzymes, is an eco-friendly and sustainable way to improve the quality of this product, while contributing to the implementation of industrial processes that have lower energy requirements and can reduce environmental impacts. This work describes the production of hydrolytic enzymes by solid-state fermentation (SSF), cultivating filamentous fungi on waste from the coffee industry, followed by their application in the hydrolysis of waste coffee polysaccharides from soluble coffee processing. Different substrate compositions were studied, an ideal microorganism was selected, and the fermentation conditions were optimized. Cultivations for enzymes production were carried out in flasks and in a packed-bed bioreactor. Higher enzyme yield was achieved in the bioreactor, due to better aeration of the substrate. The best β-mannanase production results were found for a substrate composed of a mixture of coffee waste and wheat bran (1:1 w/w), using Aspergillus niger F12. The enzymatic extract proved to be very stable for 24 h, at 50 °C, and was able to hydrolyze a considerable amount of the carbohydrates in the coffee. The addition of a commercial cellulase cocktail to the crude extract increased the hydrolysis yield by 56%. The production of β-mannanase by SSF and its application in the hydrolysis of coffee polysaccharides showed promise for improving soluble coffee processing, offering an attractive way to assist in closing the loops in the coffee industry and creating a circular economy.

Biological treatment of biomass gasification wastewater using hydrocarbonoclastic bacterium Rhodococcus opacus in an up-flow packed bed bioreactor with a novel waste-derived nano-biochar based bio-support material

Wastewater from biomass gasification chiefly contains monocyclic (MAHs) and polycyclic aromatic hydrocarbons (PAHs) that are generated by wet scrubbing process of raw synthetic gas during its cleaning process. These MAHs and PAHs are well-known as toxic, mutagenic, and carcinogenic agents which need to be removed prior to their release into the environment. The present study evaluated the potential of an up-flow packed bed bioreactor (UFPBBR) with immobilized Rhodococcus opacus onto biochar loaded polyurethane foam (PUF) as the packing material for treating biomass gasification wastewater (BGWW). Initially, the bioreactor was operated utilizing only PUF as the support material and a maximum COD removal of 81 ± 2.65% was achieved with 1820 mg L−1 of influent COD concentration and 24 h of HRT. Using biochar loaded PUF as the support material; more than 95 ± 1.27% of COD removal was attained under the same COD loading condition and HRT. Furthermore, the wastewater was detoxified to 96.2% demonstrating the ability of UFPBBR system using the novel biochar based bio-support material for biomass gasification wastewater treatment.

Valorization of Banana Peel for Citric Acid Production under Solid State Fermentation with Aspergillus niger

Valorization of banana peel (BP) through solid state fermentation (SSF) was aimed<br /> in this research. The appropriate conditions of citric acid (CA) production by SSF of<br /> Aspergillus niger were investigated. Firstly, the optimum initial pH of the BP and the<br /> aeration rate were studied by conducting SSF in a 250-mL flask and 2-L glass column,<br /> respectively. It was found that the initial pH of the BP and aeration rates affected the CA<br /> production. The results showed that the initial pH of 5.0 and 1.0 vvm aeration were appropriate for the CA production of A. niger using BP as a substrate. The problem of rising temperature during SSF was found when applying the optimum condition to the SSF<br /> in the 20-L packed bed bioreactor (PBB), which caused a decrease in the CA production<br /> compared to that of the glass column. The cooling air jacket constructed to the PBB to<br /> remove the heat during the SSF helped increase the CA production from that in the PBB.<br /> The maximum CA production in the 20-L air-jacketed PBB was 124.0±19.2 mg g–1DS.

Development of a novel fungal fluidized-bed reactor for gaseous ethanol removal.

Fluidized bed bioreactors can overcome the limitations of packed bed bioreactors such as clogging, which has been observed in the industrial application for decades. The key to establish a gaseous fluidized bed bioreactor for treatment of volatile organic compounds is to achieve microbial growth on a light packing material. In this study, Two fungal species and two bacterial species were isolated to build a fungal fluidized-bed reactor (FFBR). A light packing material with wheat bran coated on expended polystyrene was used. The FFBR was operated for 65 days for gaseous ethanol removal and obtained elimination capacities of 500-1800g∙m-3∙h-1 and removal efficiencies of 20-50%. The pressure drops was well controlled with values around 400Pa∙m-1. Stress tolerant genera including Aureobasidium, Stenotrophomonas and Brevundimonas were dominant. Meyerozyma, whose species were present in an initial inoculated isolate, was detected among the dominant species with 28.70% relative abundance; they were reported to degrade complicated compounds under similarly stressful environments.

Synthesis of 2-ethylhexyl oleate catalyzed by Candida antarctica lipase immobilized on a magnetic polymer support in continuous flow.

This study investigated the synthesis of 2-ethylhexyl oleate catalyzed by Candida antarctica lipase immobilized on magnetic poly(styrene-co-divinylbenzene) particles in a continuous packed-bed bioreactor. Runs were carried out in a solvent-free system at 50°C. The performance of the reactor was evaluated for substrates composed by oleic acid and 2-ethylhexanol at five molar ratios (1:4-4:1), determining its operation limits in terms of substrate flow rate. The system performance was quantified for three different flow rates corresponding to space-time between 3 and 12h. For each condition, the influence of the space-time in the ester formation, esterification yield and productivity was determined. The molar ratio of acid-to-alcohol interfered, in a remarkable way, in the formation of 2-ethylhexyl oleate and the best performance was attained for substrate at equimolar ratio running at 12h space-time. Under this condition, average 2-ethylhexyl oleate concentration was 471.65 ± 2.98gL-1 which corresponded to ester productivity of 23.16 ± 0.49mmolg-1L-1h-1. This strategy also gave high biocatalyst operational stability, revealing a half-life time of 2063h. A model based on the ping-pong Bi-Bi mechanism was developed to describe the kinetics of the esterification reaction and validated using experimental data. The goodness of fit of the model was satisfactory (R2 = 0.9310-0.9952).

Optimization of biodiesel synthesis from nonedible oil using immobilized bio-support catalysts in jacketed packed bed bioreactor by response surface methodology

Abstract The present study has been focussed on biodiesel production from nonedible oil in a jacketed packed bed bioreactor (JPBBR) using immobilized (lipase from Pseudomonas cepacia) biocatalyst under the cleaner production. It has no harmful by products. Experimental data were evaluated by response surface methodology (RSM) integrated desirability functional approach. RSM based on a, 4 factors and 5 levels central composite design (CCD) was employed to obtain the best possible combination which was found to be under the optimized conditions of enzymatic transesterification reactions, i.e. 2-propanol to oil molar ratio_5.93:1, time_24.32 h, enzyme (lipase) loading_9.46% (w/w) and temperature_49.7 °C. The actual experimental yield was 84.58%, which compared well to the maximum predicted value of 83.80% and conversion of biodiesel yield 85.14%. Reusability test shows the bio-support catalyst could retain up to ∼70% of its initial biodiesel yield after 10 cycles. The results showed that the immobilized lipase produced high biodiesel yield after 50 days and retained 78.58%, even after 60 days. This proves the beneficial role at industrial scale.

Surveying Denitrification Efficacy in Up-Flow Packed Bed Bioreactor Operated under Heterotrophic Condition Using Autotrophic Bacteria

Introduction: The biological denitrification process is an interesting cost-effective technique to remove nitrate from water supplies. Acetic acid can be used as a carbon source in this process, but its consumption rate is a critical issue and, in some cases, it is quite different from stoichiometric constants. The current study aimed to investigate the nitrate removal in an up-flow packed bed bioreactor. Furthermore, various parameters affecting this process were investigated and optimized. In this study, the autotrophic bacteria were used for the heterotrophic process. Materials and Methods: Initially, the autotrophic bacteria were cultured and used for the following heterotrophic conditions in distinct reactors. A pilot-scale anoxic up flow bioreactor packed was constructed using the polyethylene media and applied to remove nitrate from the aqueous environment. Consequently, the effects of hydraulic retention times (HRT) and different acetic acid concentrations as carbon source were evaluated. During the study, the amounts of alkalinity, pH, temperature, and nitrate were checked. Results: The designed bioreactor removed an average of over 88% of nitrate, while the acetic acid consumption was 2 mg/mg NO3-N, which was lower than the stoichiometric constant for heterotrophic process. Moreover, in the three studied HRTs (1.5, 3, and 5 h), the Alkalinity increased from 14.2 to 19.8 %. Conclusion: The results of this study showed high efficiency in nitrate removal via heterotrophic denitrification using acetic acid as carbon source for autotrophic bacteria.

A new thiosalt depuration bioprocess for water- recycling in metallic sulphide mineral processing

Economic and environmental imperatives force the metallic sulphide processing industry to re-use liquors that normally contain thiosalts; this processing entails difficulties in operation control and losses in metal-extraction performance. Bio-oxidation presents a promising option for thiosalt removal. Hitherto, this method has been tested to treat synthetic liquors containing only one thiosalt, either tetrathionate or thiosulphate, usually at low concentration (<1 g/L). In this article, bio-oxidation is studied of synthetic and real liquors containing mixtures of these thiosalts at concentrations close to 4.5 g/L. Coexistence of thiosulphate and tetrathionate in an acidic medium leads to strong and irreversible inhibition phenomena (performance losses close to 80%). In order to solve this problem, a previous treatment is proposed consisting of the 8-hour aeration of inlet solution after adding up to 20 mg/L and pH 1.5 of Cu2+ and sulphuric acid, respectively, resulting in the complete removal of thiosulphate and sulphite. Pre-treated real liquors were fed into a 1 L discontinuous stirred tank bioreactor and a 1 L continuous flooded packed-bed bioreactor, which were successfully started up and operated, thereby obtaining bio-oxidation rates close to the highest rates found in the literature with synthetic liquors.

Effect of hydraulic retention time and pH on oxidation of ferrous iron in simulated ferruginous acid mine drainage treatment with inoculation of iron-oxidizing bacteria

The effect of hydraulic retention time (HRT) and pH on the biooxidation of ferrous iron during simulated acid mine drainage (AMD) treatment was investigated. The simulated AMD was highly acidic (pH 2.5), rich in iron (about 1700 mg/L) and copper (about 200 mg/L), and contained high concentrations of sulfate (about 4700 mg/L). The biooxidation of ferrous iron was studied in a laboratory-scale upflow packed bed bioreactor (PBR). The HRT was shortened stepwise from 40 h to 20 h, 13 h, and 8 h under the acidic environment at a pH value of 2.2. Then, the influent pH value was changed from 2.2 to 1.2 at a constant suitable HRT. Physiochemical and microbial community structure analyses were performed on water samples and stuffing collected from the bioreactor under different conditions. The results indicate that the efficiency of ferrous iron oxidation gradually decreased with the decrease of HRT, and when the HRT exceeded 13 h, ferrous iron in AMD was almost completely oxidized. In addition, the best efficiency of ferrous iron oxidation was achieved at the influent pH value of 1.8. Microbial community structure analyses show that Leptospirillum is the predominant genus attached in the bioreactor, and low influent pH values are suitable for the growth of Leptospirillum.

Hemp Fibres as Novel Green Support Material for Immobilization of Acidithiobacillus ferrooxidans

The suitability of hemp fibres as green support material for immobilization of Acidithiobacillus ferrooxidans cells in ferrous iron bio-oxidation process was investigated. A series of experiments were carried out in shake flasks and bioreactor for both cell immobilization and iron bio-oxidation. In flask experiments, five successive batch colonization cycles resulted in 6-fold increase in ferrous iron bio-oxidation rate. Evaluation of immobilized cells activity through two approaches as wet and dry storage demonstrated the higher activity of wet stored bacteria than dry ones after a period of two months. The dry stored bacteria lost almost all their activities after eight weeks. Whereas, the activity of wet stored samples was 30% at the same time. In bioreactor experiments, use of a packed-bed bioreactor with hemp fibres, the maximum ferrous iron bio-oxidation rate of 2.46 kg m-3 h-1 was obtained at dilution rate of 3.97 h-1, and retention time of 0.25 h. Our experimental results could pave a new platform for the application of hemp fibre as an inexpensive and environmentally friendly support material in bacterial cell immobilization, especially in ferrous iron bio-oxidation process using acidophilic bacteria.

Genome-resolved metagenomics links microbial dynamics to failure and recovery of a bioreactor removing nitrate and selenate from mine-influenced water

A packed-bed bioreactor designed for selenium removal from coal waste leachate experienced reduced hydraulic conductivity and loss of performance soon after forward flow of mine-influenced water containing nitrate and selenate as the main contaminants. A metagenomic study was initiated to help diagnose the causes for the bioreactor’s poor operation. Despite being inoculated with a custom consortium of Pseudomonas strains capable of nitrate and selenate reduction, the microbial community shifted towards dominance by Trichococcus- and Candidatus Saccharibacteria- related species. Metagenomic assembled genomes for these revealed that they did not have the metabolic potential for denitrification or selenate reduction, and displayed an affinity for carbohydrate carbon sources through the PTS pathway. We speculate that the sugar-based carbon sources fed to the bioreactor lead to favourable growth of the filamentous forms of these species under the conditions of the bioreactor, which may have allowed for infiltration of oxygen. Decreasing ORP coincided with decrease in relative abundance of the filamentous species and increase in relative abundance of uncharacterized species in the Burkholderiales and Rhodocyclales orders with the metabolic potential for all steps of the denitrification pathway including nitrate reductases closely related to those with an affinity for selenate as well as nitrate.

Continuous Bioconversion of Oleuropein from Olive Leaf Extract to Produce the Bioactive Product Hydroxytyrosol Using Carrier-Immobilized Enzyme

Feasibility and stability were evaluated of a continuous multi-batch process for converting oleuropein (OLE) from olive leaf extract to the bioactive product hydroxytyrosol (HT). Carrier beads made of three different materials (calcium alginate, chitosan with deacetylated α-chitin nanofibers (DEChN), or porous ceramic) were investigated for morphology, thermogravimetric, sorption, and viscoelastic properties. Enzymatic hydrolysis of OLE conducted in a packed bed bioreactor containing cellulase immobilized to carrier beads yielded OLE degradation rates of ~ 90% and an average HT yield of ~ 70% over 20 batches. Ultimately, inorganic porous ceramic beads were less costly and exhibited superior performance relative to organic carriers and thus were deemed most suitable for industrial-scale HT production. Systems utilizing enzyme immobilization within packed bed reactors hold promise for achieving efficient production of valuable bioproducts from discarded biomass materials.

Self-build packed-bed bioreactor for rapid and effective BOD estimation.

This work demonstrated a simple, low-cost, rapid, and effective biochemical oxygen demand (BOD) estimation system based on a packed-bed bioreactor that can be easily self-built on-site at a particular wastewater treatment plant for continuous monitoring of the influent and effluent. The use of natural microbial consortium that were collected from the target wastewater and immobilized on a cheap porous carrier simply by adhesion resulted in an acceptable accuracy of over 95%. The newly developed semi-continuous operating mode with peak-type signals was shown to be able to continuously estimate BOD at a high flow rate to overcome the flow dependence of the oxygen electrode, limit clogging issues, enhance the response time, and lower the limit of detection. The resulting packed-bed bioreactors could work continuously for 22h with a coefficient of variance (CoV) of only 1.8% or for 13h a day for several days with a maximum CoV of 1.4% and their response was observed to be stable over 80 consecutive measurements. They exhibited stable responses at a wide pH range of 6.5-8.5, which is also the recommended range for aerobic wastewater treatment, emphasizing the greater ease of use of natural microorganisms for BOD estimation.

EFFECIENCY OF IMMOBILIZED POLYPHENOL OXIDASE ON SOME TEXTILE DYES DEGRADATION USING BATCH OPERATION SYSTEM BY PACKED BED BIOREACTOR

In the current study, three types of common plants, namely Tomato (Solanum lycopersicum), Cucumber (Cucumis sativus) and Orange (Citrus sinensis) were obtained and screened for their polyphenol oxidase (PPO) activity, Among the three plants, Solanum lycopersicum was chosen with maximum enzymatic activity, it had the highest productivity of the enzyme (23733 U/mg protein).The PPO from Solanum lycopersicum was purified using two steps: concentration by sucrose and gel filtration by using Sephacryl S-200. The results showed an increase in the final purification folds by 2.4 times with an enzyme yield of 32.6%. The immobilization studies showed that PPO was more stable when immobilized on chitosan by covalent linkage with immobilization ratio of 62%, in comparison with agar-agar by entrapment method (36%). The removal efficiency of crude and partial purified PPO was studied with textile dyes, including yellow, red, black and blue dyes at optimum conditions: pH 5, temperature 40oC after 3 hrs. Maximum removal efficiency of dyes observed with crude PPO were 53.9, 81.4, 86.5 and 79.6% respectively. However, purified PPO displayed removal efficiency reached 60.3, 84.3, 84.6 and 77.5% respectively. The potential of immobilized PPO on chitosan was evaluated by decolorization of black textile dye in packed bed bioreactor in batch operation. The results indicated that immobilized PPO in batch operation has the ability to remove 99% of the dye after 2 hrs, and the results showed a positive relationship between the degradation rate and incubation time in batch operation.

Modeling and validating Pseudomonas aeruginosa kinetic parameters based on simultaneous effect of bed temperature and moisture content using lignocellulosic substrate in packed-bed bioreactor

The aim of this study was to propose kinetic parameters of a bacterial growth and its products formation as functions of environmental conditions. Hence, the effect of temperature and initial moisture content on bacterial growth and rhamnolipid biosurfactant production by Pseudomonas aeruginosa were investigated in the first step. A mixture of corn bran and corn germ was used as substrate in small glass packed-bed laboratory bioreactors. Results showed the highest rhamnolipid production rate was observed in stationary phase and the highest growth and rhamnolipid production were at 35°C and initial bed moisture of 70%. Then, kinetic parameters of the bacteria were determined using the fitting of kinetic models to experimental results of cumulative CO2, rhamnolipid amount and metabolic water production. After that, the kinetic parameters mCO2, mRL, mw and YX/W were considered with constant values while μm, Xm, YX/CO2 and YRL/X were presented as functions of environmental conditions, by modifying a model and using 3-dimensional curve fitting. Finally, in order to validate the obtained kinetic parameters and functions, transport phenomena in a packed-bed bioreactor were modeled in 2-dimensions. The model results were verified by the experimental results obtained from a 3-liter glass packed-bed bioreactor and the validation was satisfactory.

Validation of a novel 3D flow model for the optimization of construct perfusion in radial-flow packed-bed bioreactors (rPBBs) for long-bone tissue engineering

Osteogenic cell culture in three-dimensional (3D) hollow cylindrical porous scaffolds in radial-flow packed-bed bioreactors (rPBBs) may overcome the transport limitations of static and axial perfusion bioreactors in the engineering of long-bone substitutes. Flow models of rPBBs help optimize radial flux distribution of medium and tissue maturation in vitro. Only a 2D model is available for steady flow transport in rPBBs with axisymmetric inlet and outlet accounting for the fluid dynamics of void spaces, assessed against literature information. Here, a novel 3D model is proposed for steady flow transport in the three compartments of rPBBs with a more practical lateral outlet. A 3D model of transient tracer transport was developed based on the flow model to predict bioreactor residence time distribution (RTD). Model-predicted flow patterns were validated in terms of RTD against tracer experiments performed with bioreactor prototypes equipped with commercial scaffolds for bone tissue engineering. Bioreactors were challenged with a step change in entering tracer concentration in an optimized set-up under conditions promoting uniform radial flux distribution and typical shunt flows. Model-predicted RTDs agreed well with those experimentally determined. In conclusion, tracer experiments validate the use of the 3D flow model for optimizing construct perfusion in rPBBs to engineer long-bone substitutes.

Transient Lentiviral Vector Production Using a Packed-Bed Bioreactor System.

Scalable lentiviral vector (LV) manufacturing is vital for successful commercialization of LV-based gene and cell therapy products. Accordingly, efforts are currently focused on developing and adapting technologies to address both upstream and downstream production bottlenecks. To overcome the limitations of current upstream processes, researchers are now favoring the use of bioreactors over traditional two-dimensional culture platforms. Bioreactors provide many advantages for manufacturing biomolecules, including process automation, tight regulation of production conditions, reduced labor input, and higher productivity potential. This study describes a transient LV production strategy employing a single-use, packed-bed bioreactor vessel. Functional LV titers in the 106 TU/mL range were achieved, and after concentration yields of up to 109 TU/mL were attained. This proof of principle study demonstrates that LV can be successfully produced in a packed-bed system. With further optimization, a packed-bed bioreactor could offer a potential scale-out solution for LV manufacturing.

Enzymatic Monolith Continuous Microreactors for the Synthesis of Xylooligosaccharides by Controlled Hydrolysis of Xylans

Immobilized enzymes are increasingly used in enzymatic microreactors, to encourage applications in different fields: food industry, medical diagnostics, organic synthesis, drug discovery, biosensors. As a matter of facts, microfluidic systems offer many advantages over conventional packed-bed bioreactors, such as higher efficiency, feasibility of multi-enzyme systems, higher mechanical stability, reduced amounts of reagents required [Pirozzi et al., 2016].In this study s-1,4-xylan obtained from hemicelluloses was used to obtain xylo-oligosaccharides (XOS), the only nutraceutical that can be produced from lignocellulosic biomass. XOS are considered non-digestible food ingredients, and their prebiotic effect of XOS on Bifidobacterium and Lactobacillus species has been recently demonstrated [Aachary et al., 2011; Broekaert et al., 2011; Nacos et al., 2006; Samanta et al, 2015; Vazquez]et al., 2000]. In addition XOS are stable at wide range of pH (2.5–8.0) and temperature (100 °C), they can stimulate the selective growth of the gut microflora, reduce the blood glucose and cholesterol, reduce the pro-carcinogenic enzymes in gastrointestinal tract, and enhance the mineral absorption from large intestine [Aachary et al., 2011; Broekaert et al., 2011; Nacos et al., 2006; Samanta et al, 2015; Vazquez]et al., 2000].In this view, sol-gel methods were used for the physical entrapment of xylanase from T. lanuginosus. Due to the mild conditions usually adopted for the gel synthesis, enzymes could be added to the initial mixture of precursors and so entrapped within monolithic porous layers in capillary tubes [Pirozzi et al., 2016].A suitable combination of sol-gel precursors was found for the simultaneous improvement of the monolith adhesion to the tubing internal surfaces and of the catalytic properties of the entrapped enzymes. Specific attention was devoted to the determination of operating conditions suitable to enhance enzyme stability as well as selectivity in terms of XOS produced.

Investigation of immobilized filamentous fungi for treatment of real textile industry wastewater using up flow packed bed bioreactor

In this study, two filamentous fungi Aspergillus carbonarius M333 and Penicillium glabrum Pg1 were immobilized in macro porous polymeric support (MPPS) to treat real textile industry wastewater. The color and chemical oxygen demand (COD) removal efficiencies were tested in a batch and continuously operated up flow packed bed bioreactor. The protein concentration was also measured for different pH and MPPS amount. Wastewater inlet pH was tested between 4.5 and 6.0 and pH 5.5 was chosen optimum working pH in order to enhance the highest color and COD removal efficiencies. Moreover, MPPS amount was also tested from 1 to 9 pieces and the highest color and COD removal efficiencies were obtained with 9 pieces. The results showed that 98.2% decolorization and 69.8% COD removal efficiencies were obtained by batch experiments. However, 78.8% decolorization and 67.7% COD removal efficiencies were achieved by up flow packed bed bioreactor. The color and COD removal efficiencies were measured 78.8%, 72.1%, 70.2% and 67.7%, 62.5%, 60.0%, respectively, when the up flow packed bed reactor was operated repeatedly for 3 cycles. The immobilized fungi on MPPS were characterized by Scanning Electron Microscopy (SEM).