Scale-up synthesis of lipase-catalyzed palm esters in stirred-tank reactor

Lipase-catalyzed production of palm esters by alcoholysis of palm oil with oleyl alcohol in n-hexane was performed in 2L stirred-tank reactor (STR). Investigation on the performance of reactor operation was carried out in batch mode STR with single impeller mounted on the centrally located shaft. Rushton turbine (RT) impellers provide the highest reaction yield (95.8%) at lower agitation speed as compared to AL-hydrofoil (AL-H) and 2-bladed elephant ear (EE) impellers. Homogenous enzyme particles suspension was obtained at 250 rpm by using RT impeller. At higher impeller speed, the shear effect on the enzyme particles caused by agitation has decreased the reaction performance. Palm esters reaction mixture in STR follows Newtons' law due to the linear relation between the shear stress (tau) and shear rate (dupsilon/dy). High stability of Lipozyme RM IM was observed as shown by its ability to be repeatedly used to give high percentage yield (79%) of palm esters even after 15 cycles of reaction. The process was successfully scale-up to 75 L STR (50 L working volume) based on a constant impeller tip speed approach, which gave the yield of 97.2% after 5h reaction time.     

Improved bacterial nanocellulose production from glucose without the loss of quality by evaluating thirteen agitator configurations at low speed

Thirteen agitator configurations were investigated at low speed in stirred-tank reactors (STRs) to determine if improved crude bacterial nanocellulose (BNC) productivity can be achieved from glucose-based media while maintaining high BNC quality using Komagataeibacter xylinus ATCC 23770 as a model organism. A comparison of five single impellers showed the pitched blade (large) was the optimal impeller at 300 rpm. The BNC production was further increased by maintaining the pH at 5.0. Among the single helical ribbon and frame impellers and the combined impellers, the twin pitched blade provided the best results. The combined impellers at 150 rpm performed better than the single impellers, and after optimizing the agitation conditions, the twin pitched blade (large) and helical ribbon impellers performed the best at 100 rpm. The performances of different agitators at low speed during BNC production were related to how efficiently the agitators improved the oxygen mass transfer coefficient. The twin pitched blade (large) was verified as providing the optimum performance by an observed crude BNC production of 1.97 g (L×d)⁻¹ and a BNC crude yield of consumed glucose of 0.41 g g⁻¹, which were 2.25 and 2.37 times higher than the initial values observed using the single impeller respectively. Further characterization indicated that the BNC obtained at 100 rpm from the STR equipped with the optimal agitator maintained high degree of polymerization and crystallinity.     

Cultivation of plant cells in a stirred vessel: effect of impeller design

Suspension cultures of Nicotiana tabacum were grown in a batch fermentor using different agitation systems. The effects of the impeller type, size, and agitation speed on the productivity of cell mass and secondary metabolites (phenolics) have been investigated. The use of a large, flat-bladed impeller (diameter 7.6 cm; width 14.0 cm) improved culture growth significantly over systems using a regular, flat-bladed impeller (diameter 5.6 cm; width 1.5 cm). An impeller of the same dimensions as the 14.0-cm-wide, large, flat-bladed impeller with sail cloth blades yielded a higher maximum growth rate in the exponential phase but resulted in a longer lag phase. Overall (intracellular and extracellular) phenolics concentration showed a direct relationship to culture growth rate whereas extracellular concentrations were a function of agitation conditions. Power consumption and flow pattern studies were also completed to further characterize the different impellers tested.     

Mixing design for enzymatic hydrolysis of sugarcane bagasse: methodology for selection of impeller configuration

One of the major process bottlenecks for viable industrial production of second generation ethanol is related with technical–economic difficulties in the hydrolysis step. The development of a methodology to choose the best configuration of impellers towards improving mass transfer and hydrolysis yield together with a low power consumption is important to make the process cost-effective. In this work, four dual impeller configurations (DICs) were evaluated during hydrolysis of sugarcane bagasse (SCB) experiments in a stirred tank reactor (3 L). The systems tested were dual Rushton turbine impellers (DIC1), Rushton and elephant ear (down-pumping) turbines (DIC2), Rushton and elephant ear (up-pumping) turbines (DIC3), and down-pumping and up-pumping elephant ear turbines (DIC4). The experiments were conducted during 96 h, using 10 % (m/v) SCB, pH 4.8, 50 °C, 10 FPU/g_biomass, 470 rpm. The mixing time was successfully used as the characteristic parameter to select the best impeller configuration. Rheological parameters were determined using a rotational rheometer, and the power consumptions of the four DICs were on-line measured with a dynamometer. The values obtained for the energetic efficiency (the ratio between the cellulose to glucose conversion and the total energy) showed that the proposed methodology was successful in choosing a suitable configuration of impellers, wherein the DIC4 obtained approximately three times higher energetic efficiency than DIC1. Furthermore a scale-up protocol (factor scale-up 1000) for the enzymatic hydrolysis reactor was proposed.     

Effect of number of turbine impellers on surface aeration in laboratory fermentor

The effect of one and two four-flat-bladed turbine impellers on the surface aeration intensity in a laboratory tormentor was investigated at different agitation speeds and sparge rates. The surface aeration was found to be generally more intensive with two impellers for the same other operating conditions because of higher power consumption and the position of the upper impeller closer to the free liquid surface. The surface aeration intensity was successfully correlated in terms of power consumption and sparge rate. The aeration number alone cannot be used for general predicting surface aeration intensities.     

Mixing studies in bioreactors

Blend times and power consumptions were determined for different arrangements of two equal diameter impellers, a high efficiency A310 and a “Dumbo Ear” impeller with three large, “elephant ear” blades designed for low shear agitation. A 9 l round-bottomed, unbaffled bioreactor was used in these studies. Blend times were taken as the time for the disappearance of the pink color of a basic solution of phenolphthalein on neutralization by excess acid, and the power consumption was obtained from torque measurements. The mixing results show that the Dumbo Ear impeller gives shorter blend times than the A310?at equal rotational speeds for most of the conditions studied. As expected, the Dumbo Ear impeller consumes more power than the A310?at the same rotational speed, due to its large area blades. However, the Dumbo Ear impeller also gives shorter blend times than the A310?at equal power consumptions.     

kLa of stirred tank bioreactors revisited

By means of improved feedback control k_La measurements become possible at a precision and reproducibility that now allow a closer look at the influences of power input and aeration rate on the oxygen mass transfer. These measurements are performed online during running fermentations without a notable impact on the biochemical conversion processes. A closer inspection of the mass transfer during cultivations showed that at least the number of impellers influences mass transfer and mixing: On the laboratory scale, two hollow blade impellers clearly showed a larger k_La than the usually employed three impeller versions when operated at the same agitation power and aeration rate. Hollow blade impellers are preferable under most operational conditions because of their perfect gas handling capacity. Mixing time studies showed that these two impeller systems are also preferable with respect to mixing. Furthermore the widths of the baffle bars depict a significant influence on the k_La. All this clearly supports the fact that it is not only the integral power density that finally determines k_La.     

Combined effect of hydrodynamic and interfacial flow parameters on lysozyme deactivation in a stirred tank bioreactor

The dynamic environment within a bioreactor and in the purification equipment is known to affect the activity and yield of enzyme production. The present research focuses on the effect of hydrodynamic flow parameters (average energy dissipation rate, maximum energy dissipation rate, average shear rate, and average normal stress) and the interfacial flow parameters (specific interfacial area and mass transfer coefficient) on the activity of lysozyme. Flow parameters were estimated using CFD simulation based on the k-epsilon approach. Enzyme deactivation was investigated in 0.1, 0.3, 0.57, and 1 m i.d. vessels. Enzyme solution was subjected to hydrodynamic stress using various types of impellers and impeller combinations over a wide range of power consumption (0.03 < P(G)/V < 7, kW/m3). The effects of tank diameter, impeller diameter, blade width, blade angle, and the number of blades on the extent of deactivation were investigated. At equal value of P(G)/V, epsilon(max), and gamma(avg), the extent of deactivation was dramatically different for different impeller types. The extent of deactivation was found to correlate well with the average turbulent normal stress and the mass transfer coefficient.     

A novel centrifugal impeller bioreactor. II. Oxygen transfer and power consumption

The effects of the impeller configuration, aeration rate, and agitation speed on oxygen transfer coefficient K(L)a were studied in a newly designed centrifugal impeller bioreactor (5-L). The oxygen transfer rates in the novel bioreactor were also compared with those in a cell-lift bioreactor with comparable dimensions. The cell-lift impeller produced much higher surface oxygen transfer rates than the centrifugal one at an agitation speed over 200 rpm. This result was in good agreement with our observation that the cell-lift impeller produced much higher unfavorable turbulence. In addition, the experiments using granulated agar particles as pseudo plant cells indicated that the K(L)a value decreased steadily with an increase in agar particle concentration, and the centrifugal impeller still demonstrated a larger K(L)a than the cell lift up to a high pseudo cell concentration of 19.5 g dry weight (DW)/L (under 150 rpm and 0.20 vvm) or 22.3 g DW/L (under 200 rpm and 0.20 vvm). Furthermore, the correlation between power number and impeller Reynolds number for both the centrifugal and the cell-lift impellers was successfully obtained, which could be used for predicting the power input required by each impeller. From the results obtained, the centrifugal impeller bioreactor is expected to have great potential in its application to shear-sensitive biological systems.     

Carbon monoxide mass transfer for syngas fermentation in a stirred tank reactor with dual impeller configurations

This study compares the power demand and gas-liquid volumetric mass transfer coefficient, kLa, in a stirred tank reactor (STR) (T = 0.211 m) using different impeller designs and schemes in a carbon monoxide-water system, which is applicable to synthesis gas (syngas) fermentation. Eleven different impeller schemes were tested over a range of operating conditions typically associated with the "after large cavity" region (ALC) of a Rushton-type turbine (D/T = 0.35). It is found that the dual Rushton-type impeller scheme exhibits the highest volumetric mass transfer rates for all operating conditions; however, it also displays the lowest mass transfer performance (defined as the volumetric mass transfer coefficient per unit power input) for all conditions due to its high power consumption. Dual impeller schemes with an axial flow impeller as the top impeller show improved mass transfer rates without dramatic increases in power draw. At high gas flow rates, dual impeller schemes with a lower concave impeller have kLa values similar to those of the Rushton-type dual impeller schemes but show improved mass transfer performance. It is believed that the mass transfer performance can be further enhanced for the bottom concave impeller schemes by operating at conditions beyond the ALC region defined for Rushton-type impellers because the concave impeller can handle higher gas flow rates prior to flooding.     

CFD simulation of mixing for high-solids anaerobic digestion

A computational fluid dynamics (CFD) model that simulates mechanical mixing for high-solids anaerobic digestion was developed. Numerical simulations of mixing manure slurry which exhibits non-Newtonian pseudo-plastic fluid behavior were performed for six designs: (i) one helical ribbon impeller; (ii) one anchor impeller; (iii) one curtain-type impeller; (iv) three counterflow (CF-2) impellers; (v) two modified high solidity (MHS 3/39°) impellers; and (vi) two pitched blade turbine impellers. The CFD model was validated against measurements for mixing a Herschel-Bulkley fluid by ribbon and anchor impellers. Based on mixing time with respect to mixing energy level, three impeller types (ribbon, CF-2, and MHS 3/39°) stand out when agitating highly viscous fluids, of these mixing with two MHS 3/39° impellers requires the lowest power input to homogenize the manure slurry. A comparison of digestion material demonstrates that the mixing energy varies with manure type and total solids concentration to obtain a given mixing time. Moreover, an in-depth discussion about the CFD strategy, the influences of flow regime and impeller type on mixing characteristics, and the intrinsic relation between mixing and flow field is included.     

Sorption of Acid Blue 9 Using Hydrilla verticillata Biomass—Optimization,Equilibrium, and Kinetics Studies

In the present study, the parameters, temperature, adsorbent dose, contact time, adsorbent size, and agitation speed were optimized for Acid Blue 9 removal from aqueous medium by Hydrilla verticillata biomass using response surface methodology (RSM). The optimum conditions for maximum removal of Acid Blue 9 from an aqueous solution of 100 mg/L were as follows: temperature 30.63°C, adsorbent dose 2.88 g/L, contact time 180 min, adsorbent size 120 mesh (0.124 mm), and agitation speed 237.39 rpm. At these optimized conditions, batch adsorption experiments were conducted to study the effect of pH and initial dye concentration for the removal of Acid Blue 9 dye. The optimum initial pH and initial dye concentration values for Acid Blue 9 removal were found to be 3.0 and 100 mg/L, respectively. Kinetic and equilibrium studies were carried out for the experimental results. From the kinetic studies it was found that pseudo-second-order kinetics suits the system well. From the equilibrium studies, the Freundlich isotherm fits the data well.     

Decolorization of Orange II dye by white-rot fungi

Agitation, temperature, inoculum size, initial pH and pH of buffered medium affected the decolorization of Orange II dye byCoriolus versicolor andFunalia trogii. The optimum temperature and initial pH value for decolorization were 30°C and 6.5–7.0, respectively; pH 4.5 was the most efficient in buffered cultures. High decolorization extents were reached at all agitation rates. At an inoculum size of more than 1 mL, the extent of decolorization changed only slightly. High extents were obtained using immobilized fungi at repeated-batch mode.     

A mathematical model for agitation-induced fragmentation of Penicillium chrysogenum

The morphology of filamentous organisms in submerged cultures varies between the pelleted and the dispersed forms depending on the strain of organism and the culture conditions. The dispersed form consists of branched and unbranched hyphae (freely dispersed form) and clumps (filamentous material in aggregates). In agitated systems, the choice of impeller geometry as well as the total power input determines the mechanical forces that might affect the morphology of filamentous species (e.g. by fragmentation) with simultaneous effects on their growth and productivity. To find out more about fragmentation of Penicillium chrysogenum caused by mechanical forces of different impeller types and agitation intensities, a population balance model has been developed. The projected area measured by image analysis was used to characterise the morphology (size) of the mycelia. In the model, the kinetics of mycelial fragmentation were expressed by a breakage rate constant K, which was assumed to be only dependent on the agitation conditions. The fragmentation rate was considered to follow a first order process in size (area) which was based on assumptions made for the mechanism of mycelial break-up, and work reported in the literature. Previously published mean and distributional data from off-line fragmentation experiments in ungassed vessels of sizes from 1.4 to 180?l were used to validate the model. For the first time a model has been found that is capable of fitting changes in mycelial morphology caused by mechanical forces generated by different impellers at various power inputs and scales. Besides the mean projected areas of the mycelia, the model allowed simulations of the projected area distributions, and changes in those distributions because of the agitation. At the small scale (1.4?l), the breakage rate constant K could be correlated well with either impeller tip speed or the “energy dissipation/circulation function”, which is based on mycelial circulation through the impeller region. The simpler but commonly used power input per unit tank volume did not correlate K adequately. The scale up data showed that only the “energy dissipation/circulation function” correlated mycelial fragmentation well. The dependence of K on biomass concentration, and its detailed dependence (if any) on the fermentation conditions at sampling, which might indicate likely breakage mechanisms, remain to be elucidated.     

Characterization of stirrers for screening studies of enzymatic biomass hydrolyses on a milliliter scale

The evaluation of mixing quality is an important factor for improving the geometry of stirred-tank reactors and impellers used in bioprocess engineering applications, such as the enzymatic hydrolysis of plant materials. Homogeneity depends on different factors, including the stirrer type and the reactor type (e.g., ratio of diameter/height, ratio of impeller tip diameter/reactor diameter) with or without baffles. This study compares two impellers for enzymatic hydrolysis of suspensions of biomass particles on a milliliter scale. Both impellers were derived from industrially relevant geometries, such as blade and grid stirrers, although the geometry of the second stirrer was slightly modified to an asymmetric shape. The stirrers were investigated with different stirrer–reactor configurations. This was done experimentally and with the aid of computational fluid dynamics. The flow field, mixing numbers, power characteristics and initial conversion rates of sugars were considered to compare the two stirrers. The simulated mixing numbers and power characteristics in baffled and unbaffled milliliter-scale reactors were found to be in good agreement with the measured mixing times and power consumption. The mixing numbers required to reach homogeneity were much higher for the symmetric impeller and remained at least twice as high as the mixing numbers required when using the asymmetric impeller. The highest initial sugar releases from milled corn stover suspensions were achieved with the asymmetric impeller shape. Regardless of the differences in the flow fields or mixing times, diverging enzymatic sugar releases could be confirmed for Newtonian media only.     

Dependence of mycelial morphology on impeller type and agitation intensity

The influence of the agitation conditions on the morphology of Penicillium chrysogenum (freely dispersed and aggregated forms) was examined using radial (Rushton turbines and paddles), axial (pitched blades, propeller, and Prochem Maxflow T), and counterflow impellers (Intermig). Culture broth was taken from a continuous fermentation at steady state and was agitated for 30 min in an ungassed vessel of 1.4-L working volume. The power inputs per unit volume of liquid in the tank, P/V(L), ranged from 0.6 to 6 kW/m(3). Image analysis was used to measure mycelial morphology. To characterize the intensity of the damage caused by different impellers, the mean total hyphal length (freely dispersed form) and the mean projected area (all dispersed types, i.e., also including aggregates) were used. [In this study, breakage of aggregates was taken into account quantitatively for the first time.]At 1.4-L scale and a given P/V(L), changes in the morphology depended significantly on the impeller geometry. However, the morphological data (obtained with different geometries and various P/V(L)) could be correlated on the basis of equal tip speed and two other, less simple, mixing parameters. One is based on the specific energy dissipation rate in the impeller region, which is simply related to P/V(L) and particular impeller geometrical parameters. The other which is developed in this study is based on a combination of the specific energy dissipation rate in the impeller swept volume and the frequency of mycelial circulation through that volume. For convenience, the function arising from this concept is called the "energy dissipation/circulation" function.To test the broader validity of these correlations, scale-up experiments were carried out in mixing tanks of 1.4, 20, and 180 L using a Rushton turbine and broth from a fed-batch fermentation. The energy dissipation/circulation function was a reasonable correlating parameter for hyphal damage over this range of scales, whereas tip speed, P/V(L), and specific energy dissipation rate in the impeller region were poor. Two forms of the energy dissipation/circulation function were considered, one of which additionally allowed for the numbers of vortices behind the blades of each impeller type. Although both forms were successful at correlating the data for the standard impeller designs considered here, there was preliminary evidence that allowing for the vortices would be valuable. (c) 1996 John Wiley & Sons, Inc.     

Comparison of biotin production by recombinant Sphingomonas sp. under various agitation conditions

Biotin production by fermentation of recombinant Sphingomonas sp./pSP304 was investigated. A complex medium containing 60g/l of glycerol and 30g/l of yeast extract was suitable for biotin production. Biotin was produced in the late logarithmic or stationary phase after glycerol starvation. The optimum pH value for biotin production was 7.0. When the dissolved oxygen concentration (DO) was controlled at a constant level, the biotin concentration produced after 120h was significantly lower than that obtained in a test tube culture. Therefore, a batchwise jar-fermentor culture with a constant agitation speed and without DO control was conducted for investigating the effect of agitation conditions on biotin production. Six types of impeller were tested: turbine-blade type, turbo-lift type, rotating mesh type (EGSTAR((R))), screw with draft tube type, Maxblend((R))type, and anchor type. With some impellers, agitation speed was also changed. Both the maximum cell concentration and biotin production varied depending on agitation conditions. Relatively high cell concentrations were attained with four of the impeller types, turbine-blade type, rotating mesh type, Maxblend((R)) type, and anchor type. Among these impellers, the turbine-blade impeller with sintered sparger was suitable for biotin production. After 120h, the cell concentration reached an OD(660) of 43 and a biotin concentration of 66mg/l was obtained, which was comparable with the results from the test tube culture. Morphological variation was also observed depending on the agitation conditions: oval-shaped, rod-shaped, and elongated-shaped cells. Biotin production was relatively high in slightly long rod-shape cells but low in elongated cells. The difference in morphology appeared to depend on the shear stress. It was found that biotin production was strongly correlated with cell length and the oxygen transfer coefficient (k(L)a); cell lengths in the range 4-7μm and k(L)a values in the range 1.5-2.0/min were found to be suitable for biotin production in jar-fermentor culture.     

Aerobic decolorization of Acid Brilliant Scarlet GR by microbial community and the community dynamics during sequencing batch processes

In this research, aerobic decolorization of Acid Brilliant Scarlet GR by microbial community was studied. Effects of conditions and dye concentraion on decolorization processes were investigated. Additionally, continuous decolorization was evaluated through sequencing batch tests and the microbial dynamics during this process was analyzed by polymerase chain reaction-denaturing gradient gel electrophoresis. The results showed that 100 mg l^?1 of the dye was completely decolorized within 12 h, which was mainly caused by biodegradation. The optimal decolorization conditions were as follows: inoculation size 2.07 g l^?1 (wet cell pellet), rotation speed 150 r min^?1, pH 5.0–7.0 and 30 °C. The processes were well described by zero-order kinetics, and more than 700 mg l^?1 of the dye would inhibit the activity of the consortium. Furthermore, the microbial community exhibited high efficiency in sequencing batch processes for continuous decolorization. Microbial community structure shifted obviously when exposed to higher concentration of the dye (500 mg l^?1), and all the dominant microorganisms were affiliated with four different phyla of Actinobacteria, Bacteroidetes, Proteobacteria and Firmicutes.     

Studies on transfer processes in mixing vessels: effect of gas on solid-liquid hydrodynamics using modified Rushton turbine agitators

The effect of gas on solid-liquid hydrodynamics in mixing vessels was studied to determine the agitation speed required to just completely suspend all the particles under gassed conditions, N _jsg and by measurement of the power consumption associated with this agitation speed. The solid particles have a mean diameter between 15–1000 μm. For their mixing are used standard and modified Rushton turbine agitators positioned singly or doubly on the same shaft. The modified turbine with a surface fraction of the perforations equal to 0.353 (TP3) was obtained through increase in the blade height of the Rushton turbine simultaneously with the perforation of the blade surface. The filled surface of the modified blade is equal to the blade surface of the standard Rushton turbine.     

Mass transfer and mixing rates in fermentation vessels

The effects on mass-transfer and overall mixing rates of varying impeller geometry and operating speed have been studied for flat-bladed turbines in laboratory fermentors, in aerated aqueous solutions, and in unaerated and aerated suspensions (1.6% w/v) of paper pulp. In the absence of suspended solid, oxygen absorption rates could be correlated directly with power input. In the pulp suspension, oxygen absorption at a given power input was influenced by impeller geometry and operating speed. The data for the three-phase system can be correlated by a dimensionless equation relating oxygen-transfer rates and mixing times to the geometrical and operating parameters of the impellers.