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Biochemical Engineering Journal 2 (1998) 197±205

Citric acid production and morphology of Aspergillus niger as functions of the mixing intensity in a stirred tank and a tubular loop bioreactor
Department of Bioscience and Biotechnology, University of Strathclyde, Glasgow G1 1XW, UK Received 20 April 1998; accepted 11 September 1998

M. Papagianni1, M. Mattey, B. Kristiansen2,*

Abstract The relationship between Aspergillus niger morphology and citric acid production was investigated in two reactor systems with different con®gurations, a tubular loop and a stirred tank bioreactor, with operating volumes of 6 and 8 dm3, respectively. Morphology was quanti®ed by image analysis. In each system, morphology, characterized by the parameter P (mean convex perimeter of clumps), and citric acid production, were agitation-dependent and closely linked. Increased agitation caused a reduction of clump sizes and results when both reactors demonstrate that the parameter P should not exceed a threshold value in order to achieve increased productivities. The results obtained from the two reactors were in agreement, both qualitatively and quantitatively. Reducing the fundamentally different mixing conditions of the two bioreactors to the order of the dimensionless mixing parameter relative mixing time ( m), results showed that the loop simulated the stirred tank. Also, relationships valid for one system accurately described the results obtained from the other system, demonstrating the validity of the relationship between morphology and productivity for the particular fermentation, regardless of the reactor type. Previous attempts to evaluate the use of loop con®gurations as scale-up tools and their performance as bioreactors, neglected the morphology of the producer micro-organisms. This study demonstrated the close link between morphology and productivity for citric acid production by A. niger, and identi®ed a morphology parameter that was used successfully to characterize the process performance. # 1998 Elsevier Science S.A. All rights reserved.
Keywords: Aspergillus niger; Morphology; Citric acid; Loop bioreactor; Relative mixing time

1. Introduction In submerged fermentations, agitation is necessary for good mass and heat transfer rates. Mechanical forces resulting from the rotation of impellers can affect ®lamentous micro-organisms in various ways, causing rate variations of growth and product formation, development of different morphological forms and breakage of ®laments. Since ®lamentous micro-organisms are of great industrial importance, the relationship between mechanical parameters, morphology and productivity has attracted the attention of many investigators [1±3]. Particular morphological forms, often agitation induced, have been considered as more suitable for certain fermentations. Several authors have noted shorter
*Corresponding author. Tel.: +47-691-18972; fax: +47-691-18610; e-mail: 1 Department of Biosystems and Agricultural Engineering, College of Engineering, University of Kentucky, Lexington, KY 40546, USA. 2 Borregaard Industries, P.O. Box 162, N-1701 Sarpsborg, Norway.

and wider cells in fungal ®laments, with greater branching frequencies and reduced aggregate sizes under conditions of high agitation [4±7]. However, optimum stirrer speeds have been reported for many processes, while, side reactions and unwanted morphologies characterized stirrer speeds out of the ranges of interest [8,9]. Studying the behavior of three citric acid producing Aspergillus niger strains, Ujcova et al. [8] observed that higher speeds resulted in thicker, densely branched ®laments, while production was optimum within a narrow range of speeds and a drop in productivity resulted from higher stirrer speeds. In recent years, much effort has focused on the discovery of new metabolites and on improvement of the respective production methods. Information is, therefore, needed about the fundamental growth behavior with respect to morphology. The characterization of the morphology of ®lamentous micro-organisms started many years ago [6]. Early attempts to link mechanical parameters with morphology and production, were limiting by the lack of the means to quantify morphology. Nowadays, methods using automated image

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when the validity of such correlations was tested under different operation scales.5 dm3 sÀ1. the circulation frequency around the impeller. downcomer and upper transverse sections was 5 cm and 2. 0. Fermentations carried out at seven different pump settings ± 100. especially built for scale-up purposes.5. circulation (tc) and mixing times (tm) were estimated and results compared on the basis of the dimensionless relative mixing time  m (tm/tc). pH 6. Process The loop reactor (APV Chemical Machinery). KH2PO4. the power dissipation rate. over a range of circulation pump settings and stirrer speeds. 200. 0.198 M.ES10 (Braun Biotech International).428. Fig.14]. / Biochemical Engineering Journal 2 (1998) 197±205 analysis permit the extraction of quantitative information on morphology parameters from ®lamentous fermentations. The mean convex perimeter of mycelial clumps served as the characteristic morphology parameter (P) to compare fermentations from the two systems. with wide use in the food and pharmaceutical industries [17]. 300.75 Â 10À3. 0. MgSO4Á7H2O. Although. the effect of agitation on citric acid production and A. spores were produced on molasses agar plates (300 kg mÀ3 cane molasses.3. Citric acid is one of the most important organic acids. In the present work. ED . 0.1. large mass transfer rates for oxygen and substrates at low energy inputs ± attracted research for its suitability as a bioreactor [15. The complexity of the problem increases when ®lamentous micro-organisms are employed.11]. 3. in terms of bulk production. Papagianni et al. non-existing in the literature. 2.14]. based on quanti®ed morphology information. respectively. 0. comparisons were based on growth kinetic parameters and production levels. NH4NO3. as in the works of van Suijdam and Metz [5]. 0. 0. For inoculum purposes. niger morphology was investigated in a stirred tank (STR) and a tubular loop (TLR) bioreactor. 18 kg mÀ3 agar. The total length was 4 m. Micro-organism An industrial strain of A. As a result. are examples of parameters which been correlated with production rates and morphology parameters. FeSO4Á7H2O. is a well known reactor type in the chemical process technology. makes scale-up an exceedingly dif®cult task in fermentation technology. The internal diameter of the culture vessel was 22 cm and the height to diameter ratio 3 : 1. was ®tted with a hydromechanical ¯ow drive (pump) and a total operating volume of 6 dm3. Materials and methods 2. with cells of different ages and subsequently different physiological stages.4. [12] and Makagiansar et al. morphology was either not given any consideration or was assessed only qualitatively. The circulation pump was situated at the lower transverse part of the loop.8) after 7 days of incubation at 308C. 1 and 1.7.1. to our knowledge. The impeller tip speed. However. niger [18]. 350. The behaviour of micro-organisms is dif®cult to predict under the constantly changing conditions in large-scale bioreactors and this.6.8. a number of interesting correlations have been published the last few years.5 Â 10À3. The loop reactor. ZnSO4Á7H2O. 1 shows a diagram of the loop reactor used in this study. Morphology in this study was quanti®ed with image analysis. 1. KCl. 0. in batch fermentations. it was either met with apparent success [2] or it was found dif®cult to prove the generality of the proposed relationships [12]. 0. In these works.5 cm for the lower transverse sections. Its characteristics ± simple design.25. 400 and 500 rpm ± which covered the entire range of pump passing frequencies practical for this reactor. Spores harvested from the plates in sterile water and the spore suspensions were used as inocula for shake ¯asks at a level of 1 Â 107 spore mlÀ1. [2]. The fermentation inoculum was 300 ml of a 40 h-old shake ¯ask culture at 308C and 200 rpm for the STR and 200 ml for the TLR. D-Glucose. The composition of the medium for shake ¯asks and bioreactor runs was as follows (kg mÀ3): 150. 150. loop reactors have been introduced as alternative fermenter designs with regard to scaleup of fermentation processes [13. the internal diameter in the riser. de®ned ¯ow paths. morphology was rarely quanti®ed and also comparable data from different reactor systems is. CaCl2Á6H2O. representing a useful tool for the design of modern model-based concepts of control engineering [10. . 2.252. The Fig. When ®lamentous micro-organisms were used. Smith et al. Since the two bioreactors had very different mixing regimes. The corresponding approximate ¯ow rates for the whole reactor were: 0. especially for the penicillin fermentation.16] and reports have shown that loop con®gurations simulated successfully stirred tanks [13. niger (PM1) was used in this study. early observations suggested links between certain morphological types and citric acid production by A. The STR was a 10 l BIOSTAT. In order to simplify the highly complex ¯ow patterns and general ¯uid dynamics in a stirred tank bioreactor. Schematic representation of the TLR. 4.

The ®ne structure of the mycelium seemed also perfectly preserved. however. to achieve a signi®cant change in response curves a larger number of titrations were required with the fermentation broth. The alkali was injected from the top of the reactor and pH measurements were made at two different placements of the pH probe. 300. two times per day for a total of 168 h. In the loop reactor the tracer (2 ml. was regarded the time required until the response oscillation was damped with all the subsequent measurements within 10% of the ®nal response. Dynamic Data Links. Foam control was by addition of polypropylene glycol (MW 2000).. Fixed samples remained unchanged for prolonged periods. In this study. [23] (13 ml 40% formaldehyde and 5 ml glacial acetic acid added to 200 ml of 50% w/v ethanol) and used for image analysis measurements by Tucker et al. The reactor was also equipped with four baf¯es. 4. 3 show the time courses for citric acid. Cambridge). was regarded the time difference between two adjacent peaks. until the response of the pulse was completely damped and each experiment was carried out in triplicate. Citric acid was determined by the method of Marier and Boulet [19] and glucose was determined with an assay kit by Boehringer Mannheim. Examination of fresh and ®xed samples showed that the ®xation method did not affect the morphology. Results and discussion In each bioreactor. 200. The CCD camera captured images of 512 Â 512 pixels. a CCD camera (Sony XC77CE). the ®rst two titrations were not considered since they did not give reproducible results. Papagianni et al. as described by Righelato et al. The length of the perimeters was measured for at least 50 clumps per sample by joining the tips of the peripheral ®laments in each clump. a PC with a frame grabber and the image analysis software (Aquitas. 3 M NaOH) was injected to the top of the downcomer section with the pH probe placed immediately below the injection point. The determination of mixing parameters tc and tm were determined using tracer response techniques. There was no difference of the estimated mixing parameters between the two. as repetitive measurements on the characteristics of vacuoles (e. Samples for assays were taken at inoculation time and thereafter two times per day to 168 h in total. 5. As mixing time. These are based on the fact that if a pulse of tracer is injected to the ¯ow a decaying sinusoidal type of response is detected downstream of the injection point [20]. as functions of the circulation pump setting (rpm) and the stirrer speed (rpm) in the TLR and the STR.M. Samples for morphological characterization were taken from 24 h. [21]. A similar procedure was used for the estimation of mixing parameters in the STR. Viscosity measurements were made on broth samples at the end of the runs (168 h) using a Ferranti model VL concentric cylinder type viscometer.1 by addition of 2 M NaOH solution. Samples were immediately ®xed with an equal volume of ®xative. 500 and 600 rpm.25 cm diameter. washing with two volumes distilled water and drying in a microwave oven (600 W) on low power for 20 min and left in a dessicator for 24 h before reweighing. The ®xed sample was further diluted with ®xative prior to measurements and the dilution was adjusted to separate the mycelial clumps on the microscope slide. The pH was not controlled in TLR fermentations. / Biochemical Engineering Journal 2 (1998) 197±205 199 agitation system consisted of three disc turbine impellers (diameter 8 cm) with six ¯at blades. no direct effect of agitation on biomass . [24]. citric acid production and morphology were clearly dependent on culture conditions. Tracer experiments carried out with the reactor ®lled with a solution of 50 mM potassium chloride in tap water (to create an adequate buffer for pH measurements) and also with the fermentation broth. Dry-weights were determined by ®ltering 20 ml of broth through preweighed glass micro®lters (grade GF/C. pH changes were recorded in both riser and downcomer sections. This was the `convex perimeter' of clumps according to the de®nition given by Paul and Thomas [11]. In both bioreactors. not taking the ®rst peak into account because of the incomplete radial distribution of the pulse during the ®rst circulation. UK).1. In all fermentations the bulk of the mycelium was in the form of clumps from the ®rst 24 h following inoculation. Six stirrer speeds were applied: 100. although in the STR the same ®nal citric acid concentrations obtained from fermentations carried out at 400. In all cases. 2 and Fig. Response curves were drawn [22] and as tc. 4. while the response was monitored on line. Quantification of morphology Morphology was quanti®ed using an image analysis system consisting of a phase contrast microscope (Olympus CH). UK). Life Science Labs. Increasing the agitation intensity citric acid production increased. Fig. respectively. 500 and 600 rpm and the operating volume was 8 dm3. A magni®cation of Â100 was used for this work and mean values were estimated for each sample. Whatman. Dissolved oxygen tension (DOT) was monitored in both reactors (Ingold steam sterilizable probe. with a grayness level from 0 to 255. 4. Since in this reactor type the air bubble distribution (through the sparger) is more even compared to the loop reactor. alkali was used as tracer and the procedure followed was based on the work of Verlaan et al. results for the mixing time were veri®ed ± to ®nd no difference ± by using a dye as a tracer and measuring the absorbance from samples taken at regular intervals.g. 400. where a single air inlet point exists. In both reactors process temperature was 288C and the air ¯ow rate 1 volume air per volume culture per minute. while in the STR it was maintained at 2. diameters and volumes) showed very good reproducibility for over a year (unpublished results).

. Research has shown that variations in DOT were not accompanied by signi®cant changes in morphology [5. There are cases where growth rates and productivities were linked to morphology. High yields were obtained without pellets. The characteristic morphology parameter P decreased as circulation frequency through the pump increased in the TLR and as stirrer speed increased in the STR.3±2. the characteristic morphology of the citric acid producing ®lament was apparent. Biomass concentrations were rather low. 5Fig. / Biochemical Engineering Journal 2 (1998) 197±205 Fig. as in the work of Belmar±Beiny and Thomas [1]. while the lowest was 5. where no link was observed. thick densely branched ®laments and it is a general belief that pelleted morphology is required for a successful citric acid fermentation [26]. as in the work of Ayazi Shamlou et al.2 kg mÀ3. 7 shows the plots of the ®nal citric acid concentrations versus the corresponding convex perimeters of clumps from fermentations carried out in both bioreactors. however. in the STR run at 300 rpm. morphology does not seem to be directly in¯uenced. Since in all fermentations in this study the DOT never fell below 40%. Gomez et al. however. Fig.8. In the STR fermentations the pH maintained at 2. 6. niger morphology responded to agitation changes and a range of different clump sizes obtained in both systems.27. each corresponding to a certain degree of agitation. The impeller damage to mycelia was also investigated by Justen et al. Many investigators have discussed effects of agitation on morphology and biosynthesis. DOT were lower ± but never fell below 40% ± at low agitation levels in both bioreactors. niger showed that no difference in morphology for pellets and ®laments could be ascribed to dissolved oxygen levels although production was enhanced by an increase in dissolved oxygen. Fig.1 within 72 h and remained unchanged thereafter. fragmentation resulted from increased shear stress applied to the mycelium was characteristic at high stirrer speeds. as discussed by Marckl et al [29]. as particles of intertwining ®laments around a small core. [31] and it was shown to be dependent on the impeller type and agitation intensity. The pH in the TLR was not controlled by titrants. In all fermentations in this study the bulk of the mycelium was in the form of clumps. Time courses for citric acid production in the STR at different stirrer speeds. Moreover. the lower yields that was obtained at low rpm could be regarded as the result of the certain morphology developed under such agitation conditions and not as the result of any mass transfer limitations. Often. when left uncontrolled it dropped to 1. for this fermentation morphology and productivity were closely linked. For the particular fermentation aeration is an important parameter for a good yield [26]. The highest biomass concentration obtained was 7. fragmentation data. It is obvious that the size of clumps should not exceed a given value in order to have maximum productivity of citric acid. levels was observed (not shown). This effect is shown in Fig.200 M. A. [28] in their work on citric acid production by A. Early studies by Clark [18]. Fig. 4 shows the characteristic morphology of the fungus at the end (168 h) of the STR run at 500 rpm. The authors showed that hyphal breakage was the dominant mechanism that determined the hyphal length of Penicillium chrysogenum under conditions of intensive agitation in 7 and 150 dm3 mechanically agitated bioreactors. Time courses for citric acid production in the TLR at different circulation pump speeds. Papagianni et al. 2. 3. while others. so.28]. Obviously this is not the case for the strain used in this study.2 kg mÀ3. showed that increased citric acid productivities were associated with a certain morphological type: small pellets and short.1 during fermentations since. in the TLR run with a pump speed of 200 rpm. not having the compact structure of what is usually referred to as pellets [25] and not macroscopically visible. since from pH 3 at the inoculation time it dropped to around 2. [30].

However. The effect of agitation on morphology in the TLR. The broader standard deviations of mean values of P at high rpm can be an indication of fragmentation (Figs. was successfully correlated with the energy dissipation/ circulation function given by the authors. . at different scales. The effect of agitation on morphology in the STR. as in the work of Smith et al. derived from mean projected areas and total hyphal lengths. Fig. 5. mean convex perimeter of clumps (P) at different stirrer speeds (168 h fermentation). 4. for example. mean ®lament length and vacuole diameters. niger in the STR at 500 rpm and 168 h of fermentation. Photograph of a mycelial clump: typical morphology of A. compared to other studies [12] and no direct effect was noticed upon the speci®c rate of citric acid production and the ®nal product yield.M. microscopic observations and plots of time courses of clump perimeters (not shown here) did not support any particular harmful pump effect on morphology [22]. [12]. In the present study. fragmentation is likely to have occurred at increased agitation levels in both bioreactors. Fig. 6. 4 and 5). the damage to ®laments should have been small. For the range of pump passing frequencies practical for the TLR. unpublished results on the timecourses of morphology parameters. mean convex perimeter of clumps (P) at different circulation pump speeds (168 h fermentation). Papagianni et al. / Biochemical Engineering Journal 2 (1998) 197±205 201 Fig. Lower productivities have been reported at high agitation rates and were attributed to cell damage. Also. suggest that fragmentation controlled morphology at stirrer speeds higher than 400 rpm.

7 while the corresponding experimentally estimated were 8. as described in the previous section. 4±6 show. 8((a) and (b)) show that on the basis of  m results from the two systems agree even quantitatively. Bo ˆ 200 corresponds to a medium longitudinal mixing while a Bo ˆ 50 to a high one. As Figs. 45. For the two bioreactors with dissimilar con®gurations and ¯ow regimes used in this study. Circulation times for the TLR (for 500±100 rpm) were: 4. As Fig. the relative mixing times calculated with the equation of Verlaan et al. 5.4.165 m3 and height 3. In loop reactors two fundamentally different mixing effects are superimposed on each other: longitudinal mixing and bachmixing due to recycling of the circulation ¯ow. investigating the in¯uence of liquid mixing on the respiration of micro-organisms in an airlift bioreactor found that the respiratory quotients were affected by the local mixing behaviour.1.2. equal to the tm divided by its corresponding tc. 14. Mixing in bioreactors determines the process performance. 6.23 m. Mixing intensity could be such a parameter. 60 and 40. This dimensionless parameter has been used by Seipenbush and Blenke [15] on studies of the effect of ¯ow pattern on productivity in yeast cultivations in a loop bioreactor. was calculated and found to be between four and eight in both bioreactors. to ®nd a parameter with which the performance of both reactor systems could be compared. while for the STR: 8. [21]. 50. The equation derived numbers were 9. The relationship between mean convex perimeter of clumps and citric acid production in the TLR and the STR. accordingly. 8(a) shows that citric acid production increases with the relative mixing time. 16 and 22 s. using the concentric cylinder viscometer in samples taken at 168 h. Papagianni et al. showed in all cases Newtonian behaviour and this can be attributed to the low biomass concentrations. only a limited number of detailed investigations on . Unfortunately. it can be expected that the mixing properties will in¯uence the morphology and ± given that morphology and production are closely linked production. It also becomes clear that agitation controls production through morphology.3. In a system where changes in agitation intensity re¯ect to the morphology of the fungus. were very close to those obtained from the experimental data. studying the axial dispersion in an airlift loop reactor with a working volume of 0. small edies. According to Blenke [16]. 8. 8(b) shows this indeed to be the case. 36. / Biochemical Engineering Journal 2 (1998) 197±205 Fig.202 M.5 and 3. the results suggest that the loop reactor simulates the STR behavior for the given conditions and scales of operation. relative mixing times in the same range strongly indicate that an equilibrium between the effects of micro and macro-mixing was the case. Verlaan et al. As also mentioned in the previous section. 6. 14 and 18 s and for the STR (for 600 to 100 rpm): 1. Both tc and tm values were smaller in the STR. 61 and 72 s. 8(a)). 10. For the tc of 4. which expresses the degree of axial mixing. For example. It would be interesting. irrespective of the reactor type. In an attempt to ®nd a parameter that brings together the results from the two systems in a form more suitable for direct comparison. In the present study. To characterize mixing. 80. Correlating the dimensionless  m with the ®nal citric acid concentrations. The model is characterized by the Bodenstein number (Bo ˆ VL/Dacca). For the characterization of ¯ow in tubular loop reactors the diffusion model has often been used [16].8 and 6 s.093 Bo. correlated the relative mixing time with the Bo to obtain the empirical correlation:  m ˆ 0. the curves obtained from the two bioreactors resemble each other and there are no fundamental differences in the results from two systems with different con®gurations. 6. tc and mixing times tm were estimated for both bioreactors. 2. the relative mixing time was used as the parameter that characterizes the mixing intensity. With the Bo obtained for the TLR in the present study. 7. Comparing the performance of the two bioreactors. 7. therefore. which were indicative of high longitudinal mixing intensities.  m. 5 and 4 and this suggests that the relative mixing time is a parameter that can be used with con®dence in characterization of mixing in the TLR. A comparison of the rheologies of fermentation broths. It appears that the amount of citric acid produced is a function of the relative mixing time and the size of clumps. 7. ¯ow through tubes is described as plug ¯ow with diffusion like disturbance caused by molecular diffusion. dead spaces and radial velocity gradients. The corresponding mixing times for the TLR were: 32. 12. Bodenstein numbers were calculated from the tracer response experimental data for the downcomer section of the loop bioreactor [22]. 15. Tubular loop reactors have been introduced in some cases as alternative fermenter designs that may have advantages with regard to scale-up of fermentation processes. the curves obtained from the two bioreactors were very similar (Fig. Fields and Slater [32]. there was no difference in the estimated mixing parameters when the tracer experiments were carried out with tap water and fermentation broth. 3. 1. 3. 10 and 18 s the corresponding Bo were 100.8. In this model. the clump perimeter should decrease as the relative mixing time increases and Fig. 40. Fig.

as Fig. it appeared that the loop behaved as the corresponding STR representing in this way a valuable tool in scale-up and down of fermentation processes. Russel et al. (5)) was formulated from the experimental data of TLR fermentations. morphology was poorly assessed and any observations made were not quanti®ed. loop bioreactors are available in the literature. Papagianni et al. (a) Citric acid concentrations and (b) mean perimeters of clumps as functions of the relative mixing time in the TLR and the STR (168 h). This ®gure suggests that lnp can be regarded as a linear function of tm described by the equation: p ˆ e3:4‡0:21m r ˆ 0:88 (1) The relationship between the morphology parameter P and lnp for the TLR may also be regarded as a linear function. The relationship between  m and lnp (p is the product of citric acid concentration kg mÀ3) for that reactor is shown in Fig. Loop bioreactor: correlation of lnp with the relative mixing time. was the test micro-organism in studies of the performance of TLR and STR. Such studies have shown that small loops can behave like stirred tanks. in the work of Kristiansen and McNeil [14]. Both ®gures show a good ®t between correlation and experimental derived points. suggesting that the correlation could describe the data points for experiments performed under different culture conditions and that the relative mixing time can be used successfully in comparison of two systems with (3) (4) Fig. Another mycelial microorganism. In both reactors cells experience similar changes in their macro and micro environment producing a response which could be independent of the reactor type. depending on the  m and P. Results from the two reactor types were comparable. were plotted against the relative mixing time and the mean convex perimeter of clumps. Growth and product formation of three metabolite producing mycelial organisms were studied and compared in TLR and STR by Ziegler et al. In Fig. tested its performance with Saccharomyces cereviciae and outlined a design procedure for a large scale tubular reactor which had considerable advantages over the more complex scale-up of a STR. . The same growth kinetics results obtained in the loop and a conventional STR. 11 and Fig. From comparative experiments in 10 and 200 dm3 STR fermentations.M. the two equations were multiplied: p2 ˆ e3:4‡0:21m e5:34À0:001P or À Á0:5 p ˆ e8:74‡0:21m À0:001P a simpler form of which is: p ˆ e4:37‡0:105m À0:005P (5) The above (Eq. Fig. / Biochemical Engineering Journal 2 (1998) 197±205 203 For this purpose a set of empirical correlations has been formulated and plotted for the TLR. along with values calculated with the Eq. 12 experimental values of lnp from both bioreactors. 8. So far. r ˆ 0:92 (2) To obtain one expression for p. Aurobasidium pullulans. however. (5) for known tm and P from STR fermentations. morphology has not been used in such comparisons and results in the present study suggest that for the particular fermentation it represents a valuable parameter in the comparison of the two reactors. [33]. 9. 10 shows. [13] designed a tubular loop reactor. 9. with the equation describing it being: p ˆ e5:34À0:001P .

The loop reactor in this study was developed as a scale-up tool [34] and the present results show it does simulate the STR behavior.204 M. the morphology parameter P (mean size of clumps) and  m. even quantitatively. experimental data and correlation derived points Fig. the results from two systems with different con®gurations were of the same trend. dependent and closely linked. 10. The above expressions are of value only for the particular strain and the considered scale of operation and for this case they support our conclusion that the loop simulates the stirred tank. the present study demonstrates how. 6. 7. niger PM1 were agitation . 11. Papagianni et al. has been stressed by many authors. the bioreactors performance compared on the basis of the dimensionless mixing parameter relative mixing time. in a fermentation in which morphology and productivity are linked. 12. different con®gurations. It appeared that the loop reactor simulates the stirred tank reactor's behavior. Loop bioreactor: correlation of lnp with the morphology parameter P (mean convex perimeter of clumps). Both. / Biochemical Engineering Journal 2 (1998) 197±205 Fig. Conclusions In each bioreactor used in the present study. were used successfully as parameters for comparison and empirical correlations derived from the loop reactor fermentations showed a good ®t when plotted along with the experimental data from stirred tank fermentations. Relationship between lnp and  m in the TLR and the STR. morphology can be used to characterize process performance. morphology and citric acid production of A. List of symbols Bo D D Dax L N p P STR TLR tc tm m V Bodenstein number (-) Impeller diameter (m) Pipe diameter (m) Axial dispersion coefficient (m2 sÀ1) length (m) Stirrer speed (rpm) Product concentration (kg mÀ3) Convex perimeter of clumps (mm) Stirred tank reactor Tubular loop reactor Circulation time (s) Mixing time (s) Relative mixing time (-) Velocity (m sÀ1) Fig. The need of quanti®ed morphology information in studies of process development and scale-up. Since. experimental data and correlation derived points. Relationship between lnp and the morphology parameter P in the TLR and the STR.

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