Countercurrent multistage fluidized bed reactor for immobilized biocatalysts: III. Hydrodynamic aspects

In Parts I and II of this series we described the modelling, design, and operation of a multistage fluidized bed reactor (MFBR) for immobilized biocatalysts. This article deals with those aspects of the MFBR which are different from single-stage fluidized beds which are operated in batch mode with respect to the solids. The semicontinuous transport of the particles requires perfect mixing of the particles in the reactor compartments, because particles are mainly transported from the bottom of these compartments. A large spread in the physical properties of the biocatalyst particles, especially of both size and density, may cause the particles to segregate into layers with different diameter and/or density. This affects the efficient use of the biocatalyst. The properties of the particles are dependent on the immobilization method. The suitability of different methods for possible future application in the MFBR is therefore compared. Because of segregation, successful use of a biofilm catalyst with a nonuniform thickness of the biofilm is doubtful. Experiments in a small scale reactor (+/- 0.1 m diameter) demonstrated that perfect particle mixing is possible using commercially available biocatalyst particles of uniform density. Co-immobilization of the biocatalyst with glass powder in a gel is a simple and effective method of increasing gel density. High density particles allow high liquid flow rates, and thus an improved external mass transfer can be achieved.The distributor plates, which separate the reactor compartments, must allow unhindered transport of particles. Therefore, the holes in these plates must have a diameter of at least 4.5 times that of the largest particles which are present in the particle mixture used. Furthermore, the plates must be designed such that, when scaling-up the reactor, a uniform liquid distribution over the cross-sectional area of the reactor occurs. Large-scale experiments were not carried out, but published correlations, indicate that particle mixing and a uniform liquid distribution can be accomplished in a large-scale reactor under similar flow conditions.     

Countercurrent multistage fluidized bed reactor for immobilized biocatalysts: I. Modeling and simulation

For the application of immobilized enzymes, fixed bed reactors are used almost exclusively. Fixed bed reactors have specific disadvantages, especially for processes with a deactivating catalyst. Therefore, we have studied a novel reactor type with continuous transport of the immobilized biocatalyst. Flow of biocatalyst is countercurrent to the substrate solution. Because of a stagewise reactor design, back-mixing of biocatalyst is very limited and transport is nearly plug flow. The reactor operates at a constant flow rate and conversion, due to constant holdup of catalytic activity. The reactor performance is compared with a configuration of fixed bed reactors. For reactions in the first-order regime, enzyme requirements in this new reactor are slightly less than for fixed bed processes. The multistage fluidized bed appears to be an attractive reactor design to use biocatalyst to a low residual activity. However, nonuniformity of the particles might affect plug flow transport of the biocatalyst. A laboratory scale reactor and experiments are described in Part II(1) of this series. Hydrodynamic design aspects of a multistage fluidized bed are discussed in more detail in Part III.(2).     

Isopropanol biodegradation by immobilized Paracoccus denitrificans in a three-phase fluidized bed reactor

A three-phase bed bioreactor including a mix of immobilized microbes was used to degrade isopropanol (IPA). The immobilization method was studied and cells immobilized with calcium alginate, polyvinyl alcohol, activated carbon, and SiO₂ were demonstrated to be the best immobilization method for the degradation of 90% of 2?g/L IPA in just 4 days, 1 day earlier than with free cells. Acetone was monitored as an indicator of microbial IPA utilization as the major intermediate of aerobic IPA biodegradation. The bioreactor was operated at hydraulic retention time (HRT) values of 32, 24, 16, 12, and 10?hr, which correspond to membrane fluxes of 0.03, 0.04, 0.06, 0.08, and 0.10?L/m²/hr, respectively. The chemical oxygen demand (COD) removal efficiencies were maintained at 98.0, 97.8, 89.1, 80.6, and 71.1% at a HRT of 32, 24, 16, 12, and 10?hr, respectively, while the IPA degradations were 98.6, 98.3, 90.3, 81.6, and 73.3%, respectively. With a comprehensive consideration of COD removal and economy, the optimal HRT was 24?hr. The results demonstrate the potential of immobilized mixed bacterial consortium in a three-phase fluidized bed reactor system for the aerobic treatment of wastewater containing IPA.     

Citric acid production by immobilized Aspergillus niger in a fluidized bed reactor

Summary Citric acid production by immobilized of Aspergillus niger in a fluidized bed reactor was performed, evaluating the productivity and the stability of the process when pulsing device was used. The application of a pulsing flow to fluidized bed reactor and the feed nitrogen limited allow to control of bioparticles morphology avoiding bed compactation. When operated at optimum pulsation frequency (0.3 s^–1) the stability of the bioreactor was maintained for more than 30 days, increasing the citric acid production in more than 52.2%.     

Operation of a three-phase biofilm fluidized sand bed reactor for aerobic wastewater treatment

A biofilm fluidized sand bed column reactor (14 L) has been operated in the three-phase mode on a soluble glucose-yeast hydrolysate substrate in which the biofilm-sand phase (1-2.5 L) was suspended by direct aeration of the bed. Within two weeks a tight biofilm was formed whose activity resulted in a 90% reduction, with loads of 10.7 kg TC/m(3)day. The residence time was 1 h. The biofilm remained intact during operation with high residence times (up to 23 h) over three weeks. Oxygen transfer coefficients varied with aeration rate and sand quantity between 0.02 and 0.04 s(-1) during non growth conditions; they decreased with increasing amounts of clean sand and were higher and relatively independent of the sand fraction with biofilm-covered sand. Aeration rates used in the 14 L reactor were 23-40 L/min (2.4-4.1 cm/s) and were sufficient to suspend 78-92% f the biofilm-covered sand. Clean sand was 50-75% suspended. Oxygen uptake rates varied between 15.4 and 23.1 mol/m(3) h.     

Degradation kinetics of phenol by immobilized cells of Candida tropicalis in a fluidized bed reactor

Degradation kinetics of phenol by free and agar-entrapped cells of Candida tropicalis was studied in batch cultures. The initial phenol degradation rate achieved with free cells was higher than that obtained with immobilized cells, when phenol concentrations up to 1000 mg l^–1 were used. However, at higher phenol concentrations, the behaviour was quite different. The initial degradation rate of the immobilized yeast cells was about 10 times higher than that of the free cells, at a phenol concentration of 3500 mg l^–1. The semicontinuous and continuous degradation of phenol by immobilized yeast cells was also investigated in a multi-stage fluidized bed reactor. The highest phenol removal efficiencies and degradation rates as well as the lowest values of residual phenol and chemical oxygen demand were obtained in the semicontinuous culture when phenol concentrations up to 1560 mg l^–1 were used.     

Application of immobilized chymotrypsin in a multistage fluidized-bed reactor

The stereospecific hydrolysis of D ,L -phenylalanine methylester with immobilized α-chymotrypsin was carried out as a model reaction for the racemate resolution of aromatic amino acids in a five staged fluidized-bed reactor (FBR). Owing to ester hydrolysis, a pH shift occurred along the reactor. Because of the pH-dependent enzyme activity a particular longitudinal pH profile had to be enforced by a proper entrance pH in order to gain an optimum conversion. In the FBR with optimum pH profile, higher conversions were achieved than in a continuous stirred tank reactor (CSTR) at the pH optimum and at the same contact time. By the application of a proton balance and the results of kinetic measurements a model was developed for the prediction of the optimum longitudinal pH profile with regard to the maximum conversion.     

Ethanol fermentation in a magnetically fluidized bed reactor with immobilized Saccharomyces cerevisiae in magnetic particles

Ethanol fermentation by immobilized Saccharomyces cerevisiae cells in magnetic particles was successfully carried out in a magnetically stabilized fluidized bed reactor (MSFBR). These immobilized magnetic particles solidified in a 2 % CaCl(2) solution were stable and had high ethanol fermentation activity. The performance of ethanol fermentation of glucose in the MSFBR was affected by initial particle loading rate, feed sugar concentration and dilution rate. The ethanol theoretical yield, productivity and concentration reached 95.3%, 26.7 g/L h and 66 g/L, respectively, at a particle loading rate of 41% and a feed dilution rate of 0.4 h(-1) with a glucose concentration of 150 g/L when the magnetic field intensity was kept in the range of 85-120 Oe. In order to use this developed MSFBR system for ethanol production from cheap raw materials, cane molasses was used as the main fermentation substrate for continuous ethanol fermentation with the immobilized S. cerevisiae cells in the reactor system. Molasses gave comparative ethanol productivity in comparison with glucose in the MSFBR, and the higher ethanol production was observed in the MSFBR than in a fluidized bed reactor (FBR) without a magnetic field.     

Hydrogen sulfide removal by immobilized Thiobacillus novellas on SiO2 in a fluidized bed reactor

The removal of hydrogen sulfide (H2S) from aqueous media was investigated using Thiobacillus novellas cells immobilized on a SiO2 carrier (biosand). The optimal growth conditions for the bacterial strain were 30 degrees C and initial pH of 7.0. The main product of hydrogen sulfide oxidation by T. novellus was identified as the sulfate ion. A removal efficiency of 98% was maintained in the three-phase fluidized-bed reactor, whereas the efficiency was reduced to 90% for the two-phase fluidized-bed reactor and 68% for the two-phase reactor without cells. The maximum gas removal capacity for the system was 254 g H2S/m3/h when the inlet H2S loading was 300 g/m3/h (1,500 ppm). Stable operation of the immobilized reactor was possible for 20 days with the inlet H2S concentration held to 1,100 ppm. The fluidized bed bioreactor appeared to be an effective means for controlling hydrogen sulfide emissions.     

Torrefaction of sawdust in a fluidized bed reactor

In the present work, stable fluidization of sawdust was achieved in a bench fluidized bed with an inclined orifice distributor without inert bed materials. A solids circulation pattern was established in the bed without the presence of slugging and channeling. The effects of treatment severity and weight loss on the solid product properties were identified. The decomposition of hemicelluloses was found to be responsible for the significant changes of chemical, physical and mechanical properties of the torrefied sawdust, including energy content, particle size distribution and moisture absorption capacity. The hydrophobicity of the torrefied sawdust was improved over the raw sawdust with a reduction of around 40 wt.% in saturated water uptake rate, and enhanced with increasing the treatment severity due to the decomposition of hemicelluloses which are rich in hydroxyl groups. The results in this study provided the basis for torrefaction in fluidized bed reactors.     

Mathematical model for the fluidized bed biofilm reactor

A mathematical model is proposed for the fluidized bed biofilm reactor (FBBR). For individual biofilm-covered particles (bioparticles) within the reactor, an analysis of intrabiofilm mass transfer and simultaneous intrinsic zero order reaction yields an effectiveness factor expression which is a function of the modified, zero order Thiele modulus, Φ_0,m. This expression is linked to a one-dimensional reactor flow model and a fluidization model to yield an overall reactor model describing convective transport and simultaneous biochemical conversion of substrate within a FBBR. For Φ_0,m<1.15, FBBR is mass transfer limited and 0.45 order kinetics are observed. For Φ_0,m<1.15, mass transfer limitations are insignificant and intrinsic zero order kinetics are observed. A sensitivity analysis using the proposed mathematical model indicates that biofilm thickness and media size are the two most important operating parameters. These two parameters can be optimized simultaneously for a specific application. The proposed model provides a rational approach for FBBR design.     

Overloading responses of a glucose-fed anaerobic fluidized bed reactor

The responses of a glucose-fed anaerobic fluidized bed (AFB) reactor towards single- and multiple-pulse overloadings are evaluated. Under the conditions tested (i.e., pulse magnitude: 3-fold, pulse frequencies: 1 and 3; and pulse durations: up to 60 h), the AFB reactor is capable of sustaining good total organic carbon (TOC) removal (i.e., >89%) during overloading periods and achieving quick recoveries (i.e., <1 week) without displaying any detrimental symptoms. This performance is a result of using porous media particles for biomass retention, which enhances the process stability by minimizing excessive washout of bacterial cells during overloading periods when biogas production intensifies. The completely mixed conditions maintained in the liquid phase also help to mitigate the overloading impacts. However, the experimental data suggest that the glucose-fed AFB reactor is more susceptible towards multiple-pulse overloadings.     

Biodegradation of sodium benzoate by a Gram-negative consortium in a laboratory-scale fluidized bed bioreactor

Gram-negative bacteria with the potential to metabolize n-alkanes and cyclic hydrocarbons were isolated from local soils and identified using 16S rDNA sequence analysis. Three isolates (CS1CO, GL1CO, GCI1CO) were identified as strains of Pseudomonas (P.) aeruginosa and a further strain (DSS2) as P. putida. Isolates were co-cultured in a laboratory-scale fluidized bed biofilm bioreactor (FBBR) utilizing sodium benzoate as the sole carbon source, under two batch and/or one continuous growth conditions. Biofilm and planktonic bacterial growth dynamics were monitored by plate counts, and optical density measurements (230 nm) determined benzoate biodegradation. Overall higher attached and planktonic bacterial counts, and benzoate depletion, were determined under batch compared to continuous conditions, and the bioreactor performed better during the second batch phase when compared to the first batch phase. It thus appeared that both the planktonic and biofilm components of the system were necessary for the most successful sodium benzoate degradation in this system.     

Biotreatment of s-triazine-containing wastewater in a fluidized bed reactor

Mixed cultures of microorganisms immobilized on sand were used to degrade s-triazine-containing industrial wastewater in a fluidized bed reactor. Immobilized cell concentrations of up to 18 g/L volatile suspended solids could be achieved with the s-triazines as sole nitrogen source for growth and carbon sources added at a C--N ratio of about 12. Maximal removal efficiencies of 80% of the s-triazines could be maintained only if (a) the bio-film thickness was limited to avoid oxygen deficiency and (b) the carbon source and complete wastewater (/=20-25 h.     

Hydrolysis of cellulose in a cellulase-bead fluidized bed reactor

Cellulase was immobilized in a collagen fibril matrix, and no leakage of cellulase from the collagen fibril matrix was observed. The immobilized cellulase was more stable than the native cellulase. The substrate cellulose was hydrolyzed quantitatively with immobilized cellulase. The final reaction product was identified as glucose. Immobilized cellulase was used in a fluidized bed reactor where the pressure drop of the fluidized bed reactor was low and constant. Cellulose was hydrolyzed to glucose by the cellulase-bead fluidized bed reactor. The minimum flow velocity (U_mf) was 0.5 cm/sec and the optimum flow velocity of the cellulose hydrolysis was 1 cm/sec.     

Biodiesel production in a magnetically-stabilized, fluidized bed reactor with an immobilized lipase in magnetic chitosan microspheres

Biodiesel production by immobilized Rhizopus oryzae lipase in magnetic chitosan microspheres (MCMs) was carried out using soybean oil and methanol in a magnetically-stabilized, fluidized bed reactor (MSFBR). The maximum content of methyl ester in the reaction mixture reached 91.3 (w/v) at a fluid flow rate of 25 ml/min and a magnetic field intensity of 150 Oe. In addition, the MCMs-immobilized lipase in the reactor showed excellent reusability, retaining 82 % productivity even after six batches, which was much better than that in a conventional fluidized bed reactor. These results suggested that a MSFRB using MCMs-immobilized lipase is a promising method for biodiesel production.     

Removal of aqueous phenol using immobilized enzymes in a bench scale and pilot scale three-phase fluidized bed reactor

The main objective of this work was to investigate the removal of aqueous phenol using immobilized enzymes in both bench scale and pilot scale three-phase fluidized bed reactors. The enzyme used in this application was a fungal tyrosinase [E.C. 1.14.18.1] immobilized in a system of chitosan and alginate. The immobilization matrix consisted of a chitosan matrix cross-linked with glutaraldehyde with an aliginate-filled pore space. This support matrix showed superior mechanical properties along with retaining the unique adsorptive characteristics of the chitosan. Adsorption of the o-quinone product by the chitosan reduced tyrosinase inactivation that is normally observed for this enzyme under these conditions. This approach allowed reuse of the enzyme in repeated batch applications. For the bench scale reactor (1.2-l capacity) more than 92% of the phenol could be removed from the feed water using an immobilized enzyme volume of 18.5% and a residence time of the liquid phase of 150 min. Removal rates decreased with subsequent batch runs. For the pilot scale fluidized bed (60 l), 60% phenol removal was observed with an immobilized enzyme volume of 5% and a residence time of the liquid phase of 7 h. Removal decreased to 45% with a repeat batch run with the same immobilized enzyme.     

Continuous production of chiral 1,3-butanediol using immobilized biocatalysts in a packed bed reactor: promising biocatalysis method with an asymmetric hydrogen-transfer bioreduction

An asymmetric hydrogen-transfer biocatalyst consisting of mutated Rhodococcus phenylacetaldehyde reductase (PAR) or Leifsonia alcohol dehydrogenase (LSADH) was applied for some water-soluble ketone substrates. Among them, 4-hydroxy-2-butanone was reduced to (S)/(R)-1,3-butanediol, a useful intermediate for pharmaceuticals, with a high yield and stereoselectivity. Intact Escherichia coli cells overexpressing mutated PAR (Sar268) or LSADH were directly immobilized with polyethyleneimine or 1,6-diaminehexane and glutaraldehyde and evaluated in a batch reaction. This system produced (S)-1,3-butanediol [87% enantiomeric excess (e.e.)] with a space time yield (STY) of 12.5 mg h⁻¹ ml⁻¹ catalyst or (R)-1,3-butanediol (99% e.e.) with an STY of 60.3 mg h⁻¹ ml⁻¹ catalyst, respectively. The immobilized cells in a packed bed reactor continuously produced (R)-1,3-butanediol with a yield of 99% (about 49.5 g/l) from 5% (w/v) 4-hydroxy-2-butanoate over 500 h.