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1.
The gassing out method was used to study the effect of Pluronic F-68 on oxygen transport in an agitated, sparged bioreactor. At a low air sparge rate (0.07 vvm), the addition of Pluronic F-68 had a minimal effect on KLa in both water and medium at low agitation rates. As the agitation rate was increased at this low sparge rate the relative inhibitory effect of Pluronic F-68 on KLa increased with increasing agitation rate. At an air sparge rate of 0.80 vvm, the addition of Pluronic F-68 resulted in a significant reduction in KLa at all agitation rates studied.  相似文献   

2.
Gas–liquid mass transfer is often rate‐limiting in laboratory and industrial cultures of aerobic or autotrophic organisms. The volumetric mass transfer coefficient kLa is a crucial characteristic for comparing, optimizing, and upscaling mass transfer efficiency of bioreactors. Reliable dynamic models and resulting methods for parameter identification are needed for quantitative modeling of microbial growth dynamics. We describe a laboratory‐scale stirred tank reactor (STR) with a highly efficient aeration system (kLa ≈ 570 h?1). The reactor can sustain yeast culture with high cell density and high oxygen uptake rate, leading to a significant drop in gas concentration from inflow to outflow (by 21%). Standard models fail to predict the observed mass transfer dynamics and to identify kLa correctly. In order to capture the concentration gradient in the gas phase, we refine a standard ordinary differential equation (ODE) model and obtain a system of partial integro‐differential equations (PIDE), for which we derive an approximate analytical solution. Specific reactor configurations, in particular a relatively short bubble residence time, allow a quasi steady‐state approximation of the PIDE system by a simpler ODE model which still accounts for the concentration gradient. Moreover, we perform an appropriate scaling of all variables and parameters. In particular, we introduce the dimensionless overall efficiency κ, which is more informative than kLa since it combines the effects of gas inflow, exchange, and solution. Current standard models of mass transfer in laboratory‐scale aerated STRs neglect the gradient in the gas concentration, which arises from highly efficient bubbling systems and high cellular exchange rates. The resulting error in the identification of κ (and hence kLa) increases dramatically with increasing mass transfer efficiency. Notably, the error differs between cell‐free and culture‐based methods of parameter identification, potentially confounding the determination of the “biological enhancement” of mass transfer. Our new model provides an improved theoretical framework that can be readily applied to aerated bioreactors in research and biotechnology. Biotechnol. Bioeng. 2012; 109: 2997–3006. © 2012 Wiley Periodicals, Inc.  相似文献   

3.
Summary A twin bubble column was used to measure the kLa values for oxygen in model and cultivation media using the steady state method described previously (Adler et al. 1980). Desmophen and soy oil were used as antifoam agents together with model and/or cultivation media for Chaetomium cellulotyticum, Trichoderma reesei, Hansenula polymorpha, Saccharomyces cerevisiae and Escherichia coli. The bubble coalescence behavior is mainly influenced by antifoam agents and somewhat by protein and alcohol additives. In the range investigated (0.01 to 0.1%.), the kLa values are not influenced by the Desmophen concentration and only slighthly by the soy oil concentration (0.5 to 1.5%.). The coalescence behaviour was characterized by the ratio mcorr=(kLa)corr/(kLa)ref. A nutrient salt solution with Desmophen was used as a reference. The kLa measured in the investigated media were corrected by considering the differences in kLa's in the investigated and reference media. These mcorr values can directly be used for bubble columns close to the optimum aeration rate.Symbols a Specific gas/liquid interfacial area - c Concentration - kL Mass transfer coefficient - kLa Volumetric mass transfer coefficient - WSG Superficial gas velocity - EG Relative gas hold-up  相似文献   

4.
The calculation and scale-up of fermentation processes need kLa as one of the most important engineering data. There are two methods to determine kLa depending on power input, aeration rate and the properties of the fermentation broth: static with a balance between air supply and exit, dynamic gassing out with following the changes of dissolved oxygen concentration during periods of air off and a following air on. Within early intervals of fermentation time the data from both methods agree well, while for later time intervals the dynamic method always gives much lower values for kLa than static. The only explanations for this phenomenon are quick changes in the oxygen metabolism or an enzymatic storage of oxygen. For both gassing out and saturation period it is possible to calculate the same absolute amounts of this additional oxygen.  相似文献   

5.
The effects of superficial gas velocity (Ugr), gas entrance velocity (ν), and bubble size on the growth of Tisochrysis lutea was investigated in 600-mL photobioreactors operated with airlift pumps. Superficial gas velocities, calculated from measured air flow rates, ranging from 7 to 93 mm s?1 were created using a 1.6-mm diameter syringe. We tested the effects of sparger velocity over a range of 2.48 to 73.4 m s?1 and the effects of bubble size by using two styles of air stones and an open glass pipette, which created a bubble sizes in the range of 0.5 to 5 mm. We calculated oxygen mass transfer coefficient, kLa, values for all experimental conditions. Cell growth increased linearly with increased superficial gas velocity and decreased with increased sparger velocity. Results indicated that smaller bubble size leads to some initial cell damage, but after time, the increased gas transfer as reflected by the kLa value produced higher growth than larger bubbles. Two mechanisms were observed to correlate with cell damage in T. lutea: increasing velocity at the sparger tip and bubble bursting at the surface. These results demonstrate a method to test sensitivity of T. lutea to aeration, which is important for the design of airlift systems.  相似文献   

6.
Orbitally shaken cylindrical bioreactors [OrbShake bioreactors (OSRs)] without an impeller or sparger are increasingly being used for the suspension cultivation of mammalian cells. Among small volume OSRs, 50‐mL tubes with a ventilated cap (OSR50), originally derived from standard laboratory centrifuge tubes with a conical bottom, have found many applications including high‐throughput screening for the optimization of cell cultivation conditions. To better understand the fluid dynamics and gas transfer rates at the liquid surface in OSR50, we established a three‐dimensional simulation model of the unsteady liquid forms (waves) in this vessel. The studies verified that the operating conditions have a large effect on the interfacial surface. The volumetric mass transfer coefficient (kLa) was determined experimentally and from simulations under various working conditions. We also determined the liquid‐phase mass transfer coefficient (kL) and the specific interfacial area (a) under different conditions to demonstrate that the value of a affected the gas transfer rate more than did the value of kL. High oxygen transfer rates, sufficient for supporting the high‐density culture of mammalian cells, were found. Finally, the average axial velocity of the liquid was identified to be an important parameter for maintaining cells in suspension. Overall these studies provide valuable insights into the preferable operating conditions for the OSR50, such as those needed for cell cultures requiring high oxygen levels. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:192–200, 2017  相似文献   

7.
Despite the increasing importance of airlift fermentors, very little published information is available on how the geometric configurations of the draft tubes and the air-sparging system affect the mixing and oxygen transfer characteristics of the fermentor. A 14-L air-lift fermentor was designed and build with a fixed liquid height to diameter ratio of 1.5 utilizing four equally spaced air jets at the bottom. Two jet orifice sizes were used, 1.27 and 3.81 mm i.d., and for each jet size the following four geometric configurations were used: Single inner concentric draft tube, single outer concentric draft tube, two concentric draft tubes, and no draft tubes where the fermentor was operated as a shallow bubble column. It was found that the presence of draft tubes stabilized liquid circulation patterns and gave systemically higher mixing times than those obtained in the absence of draft tubes. In addition, the double draft tube geometry resulted in higher mixing times than the single draft tubes. For the power unit volume range 20 to about 250 W/m3 the larger 3.81-mm orifices gave systemically higher kL a values than the smaller 1.27-mm i.d. orifices. At 200 W/m3 the use of a single outer draft tube with the 3.81-mm orifices resulted in 94% increase in kL a values over that obtained with no draft tubes. However, the effect of draft tube geometry on kL a values when the 1.27-mm orifices were used was not significant. The air bubble formation characteristics at the jet orifices were found to be different, which reflected the differences observed in mass transfer and mixing characteristics. The power economy for oxygen transfer was found to be depend strongly on the orifice size and less on the geometric configuration of draft tubes.  相似文献   

8.
During the past decade, novel disposable cell culture vessels (generally referred to as Process Scouting Devices or PSDs) have become increasingly popular for laboratory scale studies and seed culture generation. However, the lack of engineering characterization and online monitoring tools for PSDs makes it difficult to elucidate their oxygen transfer capabilities. In this study, a mass transfer characterization (kLa) of sensor enabled static and rocking T‐flasks is presented and compared with other non‐instrumented PSDs such as CultiFlask 50®, spinner flasks, and SuperSpinner D 1000®. We have also developed a mass transfer empirical correlation that accounts for the contribution of convection and diffusion to the volumetric mass transfer coefficient (kLa) in rocking T‐flasks. We also carried out a scale‐down study at matched kLa between a rocking T75‐flask and a 10 L (2 L filling volume) wave bioreactor (Cultibag®) and we observed similar DO and pH profiles as well as maximum cell density and protein titer. However, in this scale‐down study, we also observed a negative correlation between cell growth and protein productivity between the rocking T‐flask and the wave bioreactor. We hypothesize that this negative correlation can be due to hydrodynamic stress difference between the rocking T‐flask and the Cultibag. As both cell culture devices share key similarities such as type of agitation (i.e., rocking), oxygen transfer capabilities (i.e., kLa) and disposability, we argue that rocking T‐flasks can be readily integrated with wave bioreactors, making the transition from research‐scale to manufacturing‐scale a seamless process. Biotechnol. Bioeng. 2012;109: 2295–2305. © 2012 Wiley Periodicals, Inc.  相似文献   

9.
For multiple-use bench scale and larger bioreactors, sintered stainless steel frit spargers are commonly used as microspargers. For bench-scale single-use bioreactors (SUBs), existing microspargers are sintered plastics, such as polyethylene. However, though plastics are readily sterilized by irradiation making them convenient for single use, these designs overlook surface energy properties of the materials of construction. For these sintered plastic spargers, forces at the water-air-surface interface cause bubble coalescence, leading to lower effective mass transfer, higher gas flow rates, and differing pCO2 profiles in cell culture. Alternative materials of construction were evaluated based on contact angle information and bubble formation observations. Sintered glass was chosen over thermoplastic polymers for higher surface wettability as described in the glass/water contact angle, its history as a commonly sintered material, and availability at costs suitable for single use applications. Glass sintered spargers and traditional stainless steel frit spargers were compared by porosity, bubble size, and kLa studies. Mass transfer (kLa) and cell culture performance equal or greater than a standard 20 μm stainless steel microsparger mass transfer efficiency was achieved by a glass frit sparger, of international porosity standard “P40” according to ISO 4793-80, which corresponds to a range of 16–40 μm.  相似文献   

10.
The promotory effects have been studied of the non‐ionic surfactant, Pluronic F‐68, on bud induction/shoot regeneration in epicotyl and cotyledon explants of Citrus depressa and on shoot regeneration from leaf segments of 4–6 week‐old axenic nodal segment‐derived in vitro plants of Passiflora mollissima, P. giberti and P. edulis var. flavicarpa. For epicotyls of C. depressa, supplementation of agar‐solidified MS‐based bud induction/shoot regeneration medium with 0.5% [w/v] Pluronic F‐68 significantly (P < 0.05) increased mean fresh weight gain of cultures, percentage of explants giving shoots and number of shoots per explant. The same Pluronic concentration also enhanced the mean percentage of cotyledons exhibiting bud induction and the number of buds regenerated per cotyledon explant. Fresh weight gain was unaffected across the range of concentrations (0.001–0.5% w/v) of Pluronic F‐68 evaluated for this latter explant source. For leaf explants from axenic shoot cultures of P. mollissima, supplementation of NN‐based medium, containing 3 mg/l 6‐benzyladenine and 2.0 mg/l kinetin with 0.001–0.5% [w/v] Pluronic F‐68, significantly (P < 0.05) increased mean (± s.e.m.) biomass gain by a maximum of 2.7 ± 0.1 g fresh weight (g f.wt.) over the control. Similarly, for leaf explants of P. giberti, 0.001–0.5% [w/v] Pluronic F‐68 in MS‐based medium, containing 1.0 mg/l 6‐BAP and 0.5 mg/l kinetin significantly (P < 0.05) increased mean percentage of explants undergoing shoot regeneration. For P. edulis leaf explants, mean f.wt. gain was also significantly (P < 0.05) higher with Pluronic F‐68 at 0.001–0.5% [w/v].  相似文献   

11.
Oxygen and substrate supply have always been considered physical constraints for the performance and operation of two-phase partitioning bioreactors (TPPB), widely used for the degradation of hydrophobic substrates. In this regard, the potential advantages of static mixers in upgrading the oxygen transfer and liquid-liquid dispersions in TPPB have been highlighted. In the present paper, the concomitant influence of static mixers on the gas-liquid mass transfer coefficient k L a and on substrate bioavailability was examined in TPPB. The static method based on conventional forms was developed to estimate the oxygen volumetric mass transfer coefficient. Over a broad range of liquid and air flow rates, the presence of static mixers was found to significantly enhance k L a relative to a mixer-free mode of operation. For identical conditions, static mixers improved the k L a threefold. In the presence of external aeration supply, the boost in the k L a was associated with an increase of 16% in the phenanthrene biodegradation rate due to bubble break up accomplished by the static mixers. On the other hand, static mixers were efficient in enhancing substrate bioavailability by improving the liquid-liquid interfacial area. This effect was reflected by a threefold increase in the degradation rate in the bioreactors with no external supply of air when equipped with static mixers.  相似文献   

12.
Lignocellulosic biomass such as agri‐residues, agri‐processing by‐products, and energy crops do not compete with food and feed, and is considered to be the ideal renewable feedstocks for biofuel production. Gasification of biomass produces synthesis gas (syngas), a mixture primarily consisting of CO and H2. The produced syngas can be converted to ethanol by anaerobic microbial catalysts especially acetogenic bacteria such as various clostridia species.One of the major drawbacks associated with syngas fermentation is the mass transfer limitation of these sparingly soluble gases in the aqueous phase. One way of addressing this issue is the improvement in reactor design to achieve a higher volumetric mass transfer coefficient (kLa). In this study, different reactor configurations such as a column diffuser, a 20‐μm bulb diffuser, gas sparger, gas sparger with mechanical mixing, air‐lift reactor combined with a 20‐μm bulb diffuser, air‐lift reactor combined with a single gas entry point, and a submerged composite hollow fiber membrane (CHFM) module were employed to examine the kLa values. The kLa values reported in this study ranged from 0.4 to 91.08 h?1. The highest kLa of 91.08 h?1 was obtained in the air‐lift reactor combined with a 20‐μm bulb diffuser, whereas the reactor with the CHFM showed the lowest kLa of 0.4 h?1. By considering both the kLa value and the statistical significance of each configuration, the air‐lift reactor combined with a 20‐μm bulb diffuser was found to be the ideal reactor configuration for carbon monoxide mass transfer in an aqueous phase. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2011  相似文献   

13.
Bioreactors are of interest for gas-to-liquid conversion of stranded or waste industrial gases, such as CO, CH4, or syngas. Process economics requires reduction of bioreactor cost and size while maintaining intense production via rapid delivery of gases to the liquid phase (i.e., high kLa). Here, we show a novel bioreactor design that outperforms all known technology in terms of gas transfer energy efficiency (kLa per power density) while operating at high kLa (i.e., near 0.8 s−1). The reactor design uses a micro-jet array to break feedstock gas into a downward microbubble flow. Hydrodynamic and surfactant measurements show the reactor's advanced performance arises from its bubble breakage mechanism, which limits fluid shear to a thin plane located at an optimal location for bubble breakage. Power dissipation and kL are shown to scale with micro-jet diameter rather than reactor diameter, and the micro-jet array achieves improved performance compared to classical impinging-jets, ejector, or U-loop reactors. The hydrodynamic mechanism by which the micro-jets break bubbles apart is shown to be shearing the bubbles into filaments then fragmentation by surface tension rather than “cutting in half” of bubbles. Guided by these hydrodynamic insights, strategies for industrial design are given. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2710, 2019  相似文献   

14.
The sufficient provision of oxygen is mandatory for enzymatic oxidations in aqueous solution, however, in process optimization this still is a bottleneck that cannot be overcome with the established methods of macrobubble aeration. Providing higher mass transfer performance through microbubble aerators, inefficient aeration can be overcome or improved. Investigating the mass transport performance in a model protein solution, the microbubble aeration results in higher kLa values related to the applied airstream in comparison with macrobubble aeration. Comparing the aerators at identical kLa of 160 and 60 1/h, the microbubble aeration is resulting in 25 and 44 times enhanced gas utility compared with aeration with macrobubbles. To prove the feasibility of microbubbles in biocatalysis, the productivity of a glucose oxidase catalyzed biotransformation is compared with macrobubble aeration as well as the gas‐saving potential. In contrast to the expectation that the same productivities are achieved at identically applied kLa, microbubble aeration increased the gluconic acid productivity by 32% and resulted in 41.6 times higher oxygen utilization. The observed advantages of microbubble aeration are based on the large volume‐specific interfacial area combined with a prolonged residence time, which results in a high mass transfer performance, less enzyme deactivation by foam formation, and reduced gas consumption. This makes microbubble aerators favorable for application in biocatalysis.  相似文献   

15.

Miniaturized bubble columns (MBCs) have different hydrodynamics in comparison with the larger ones, but there is a lack of scientific data on MBCs. Hence, in this study, the effect of gas hold-up, flow regimes, bubble size distribution on volumetric oxygen mass transfer coefficient at different pore size spargers and gas flow rates in MBCs in the presence and absence of microorganisms were investigated. It was found that flow regime transition occurred around low gas flow rates of 1.18 and 0.85 cm/s for small (16–40 µm) and large (40–100 µm) pore size spargers, respectively. Gas hold-up and KLa in MBC with small size sparger were higher than those with larger one, with an increasing effect in the presence of microorganisms. A comparison revealed that the wall effect on the flow regime and gas hold-up in MBCs was greater than bench-scale bubble columns. The KLa values significantly increased up to tenfold using small pore size sparger. In the MBC and stirred tank bioreactors, the maximum obtained cell concentrations were OD600 of 41.5 and 43.0, respectively. Furthermore, it was shown that in MBCs, higher KLa and lower turbulency could be achieved at the end of bubbly flow regime.

  相似文献   

16.
Summary Volumetric mass transfer coefficients (kLa) were measured by a steady state method in a twin bubble column to characterize the coalescence behaviour of the medium. Employing Hansenula polymorpha cultivation broths, kLa values were compared with those measured in model media in the presence and absence of antifoam agents. The ratio of the volumetric mass transfer coefficient in the system investigated to that in water, , was employed to characterize the cultivation medium.Symbols a Specific gas/liquid interfacial area with regard to the liquid volume in reactor - de Dynamical equilibrium bubble diameter - dH Perforated plate hole diameter - dp Primary bubble diameter - dS Sauter bubble diameter - Fv Liquid feed rate - H Bubbling layer height - kL Gas/liquid mass transfer coefficient - kLa Volumetric mass transfer coefficient - m kLa/(kLa)r coalescence index - mcorr Corrected coalescence index [Eq. (3)] - OTR Oxygen transfer rate - PO Dissolved O2-partial pressure in BS2 - P1 Dissolved O2-partial pressure in BS1 - PO PO/PS relative oxygen saturation in BS2 - P1 P1/PS relative oxygen saturation in BS1 - PS Saturation dissolved oxygen partial pressure - Rc dnB/dt coalescence rate - S Substrate concentration - tF Time since the beginning of the cultivation - X Biomass concentration - V1 Liquid volume in BS1 - wSG Superficial gas velocity in BS1 - G Gas holdup in BS1 - 1 V1/Fv mean liquid residence time in BS1 - BS1 O2 absorber column - BS2 O2 desorber column - D Desmophen (antifoam agent) - NS Nutrient salt solution (Table 1)  相似文献   

17.
Orbitally shaken bioreactors (OSRs) are commonly used for the cultivation of mammalian cells in suspension. To aid the geometry designing and optimizing of OSRs, we conducted a three‐dimensional computational fluid dynamics (CFD) simulation to characterize the flow fields in a 10 L cylindrical OSR with different vessel diameters. The liquid wave shape captured by a camera experimentally validated the CFD models established for the cylindrical OSR. The geometry size effect on volumetric mass transfer coefficient (kLa) and hydromechanical stress was analyzed by varying the ratio of vessel diameter (d) to liquid height at static (hL), d/hL. The highest value of kLa about 30 h?1 was observed in the cylindrical vessel with the d/hL of 6.35. Moreover, the magnitudes of shear stress and energy dissipation rate in all the vessels tested were below their minimum values causing cells damage separately, which indicated that the hydromechanical‐stress environment in OSRs is suitable for cells cultivation in suspension. Finally, the CFD results suggested that the d/hL higher than 8.80 should not be adopted for the 10 L cylindrical OSR at the shaking speed of 180 rpm because the “out of phase” state probably will happen there.  相似文献   

18.
Summary Hansenula polymorpha was cultivated in a bubble column loop bioreactor employing ethanol and/or glucose as substrates. By varying the substrate concentration, the cultivations were carried out in non-limited, substrate limited and oxygen transfer limited growth ranges. The influence of the transitions from one range to another on reactor performance (OTR,k L a, a) and cell productivity () were investigated. When employing ethanol as a substrate, the concentration considerably influences the fluid dynamics, mass transfer phenomena and cell productivity. When employing glucose as a substrate, glucose repression occurs. At high glucose concentrations no transition into the oxygen transfer limited growth is possible. The ethanol produced during the glucose repression influences the fluid dynamics, mass transfer phenomena and productivity. With decreasing glucose concentration the glucose repression can be gradually eliminated.  相似文献   

19.
Sugarcane bagasse is a low-cost and abundant by-product generated by the bioethanol industry, and is a potential substrate for cellulolytic enzyme production. The aim of this work was to evaluate the effects of air flow rate (Q AIR), solids loading (%S), sugarcane bagasse type, and particle size on the gas hold-up (ε G) and volumetric oxygen transfer coefficient (k L a) in three different pneumatic bioreactors, using response surface methodology. Concentric tube airlift (CTA), split-cylinder airlift (SCA), and bubble column (BC) bioreactor types were tested. Q AIR and  %S affected oxygen mass transfer positively and negatively, respectively, while sugarcane bagasse type and particle size (within the range studied) did not influence k L a. Using large particles of untreated sugarcane bagasse, the loop-type bioreactors (CTA and SCA) exhibited higher mass transfer, compared to the BC reactor. At higher  %S, SCA presented a higher k L a value (0.0448 s?1) than CTA, and the best operational conditions in terms of oxygen mass transfer were achieved for  %S < 10.0 g L?1 and Q AIR > 27.0 L min?1. These results demonstrated that pneumatic bioreactors can provide elevated oxygen transfer in the presence of vegetal biomass, making them an excellent option for use in three-phase systems for cellulolytic enzyme production by filamentous fungi.  相似文献   

20.
Conversion of D‐xylose to xylitol by Candida boidinii NRRL Y‐17213 was studied under anaerobic and oxygen limited conditions by varying the oxygen transfer coefficient kLa. Shake flask experiments were used to provide the preliminary information required to perform experiments in a bioreactor. The yeast did not grow under fully anaerobic conditions, but anaerobic formations of xylitol, ethanol, ribitol, and glycerol were observed as well as D‐xylose assimilation of 11 %. In shake flasks, with an initial D‐xylose concentration of 50 g/L, an increase in kLa from 8 to 46 h–1 resulted in a faster growth, higher rate of substrate uptake and lower yields of products. The highest xylitol productivity (0.052 g/L h) was attained at kLa = 8 h–1. At kLa = 46 h–1, 98.6 % of D‐xylose was consumed and mainly converted to biomass. Using 130 g/L D‐xylose, kLa was varied in the fermenter from 26 to 78 h–1. The percentage of consumed D‐xylose increased from 31 % at kLa = 26 h–1 to 93–94 % at all other aeration levels. Biomass yield increased with kLa, whereas ethanol, ribitol, and glycerol yields exhibited an opposite dependence on the oxygenation level. The most favorable oxygen transfer coefficient for xylitol formation, in the fermenter, was kLa = 47 h–1 when its concentration (57.5 g/L) surpassed ethanol accumulation by 3.6‐fold, and the glycerol plus ribitol by 10‐fold. Concurrently, xylitol yield and productivity reached 0.45 g/g and 0.26 g/L h, respectively. The volumetric xylitol productivity was affected more by changes in the aeration than the corresponding yield.  相似文献   

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