A dense cell culture system for aerobic microorganisms using a shaken ceramic membrane flask with surface aeration

Kamoshita, Yuya; Ohashi, Ryo; Suzuki, Takahiro

doi:10.1016/s0922-338x(97)86771-2

Cited by 12 publications

(5 citation statements)

References 5 publications

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“…The accumulation of volatile components in the reaction medium, on the other hand, may be a problem in the shaking¯ask, since the gas exchange due to diffusive mass transfer through the plug closure can be much lower in the shaking¯ask than in the stirred-tank reactor; see also [7,8].…”

Section: Discussionmentioning

confidence: 99%

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Process-engineering characterization of small-scale bubble columns for microbial process development

Weuster‐Botz

Altenbach-Rehm

Hawrylenko³

2001

Bioprocess and Biosystems Engineering

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Volumetric oxygen transfer rates and power inputs were estimated by a model of the formation of primary gas bubbles at the static sparger (sinter plate) of small-scale bubble columns and a common mass-transfer correlation for bubbles rising in a non-coalescent Newtonian electrolyte solution of low viscosity. Estimations were used to assess the dimensioning and possibilities of smallscale bubble column application with an height/diameter ratio of about 1. Estimations of volumetric oxygen transfer rates (<0.16 s ±1 ) and power inputs (<100 W m ±3 ) with a mean pore diameter of the static sparger of 13 lm were con®rmed as function of the super®cial air velocity (<0.6 cm s ±1 ) by measurements using an Escherichia coli fermentation medium. Small-scale bubble columns are thus to be classi®ed between shaking¯asks and stirredtank reactors with respect to the oxygen transfer rate, but the maximum volumetric power input is more than one magnitude below the power input in shaking¯asks, which is of the same order of magnitude as in stirred-tank reactors. A small-scale bubble columns system was developed for microbial process development, which is characterized by handling in analogy to shaking¯asks, high oxygen transfer rates and simultaneous operation of up to 16 small-scale reactors with individual gas supply in an incubation chamber. List of symbolsA cross-sectional area of the sparger (m 2 ) d b primary bubble diameter (m) d b Ã equilibrium bubble diameter (m) D O 2 diffusion coef®cient of oxygen in the liquid phase (m 2 s ±1 ) d p pore diameter of the static sparger (m) f correlation factor g acceleration due to gravity (m s ±2 ) F r surface tension force at the opening of a pore of the static sparger (N) F g resistance force (N) F A buoyancy force of the bubble averaged over the time of the bubbling process (N) F g gas¯ow rate (m 3 s ±1 ) F g,p gas¯ow rate in the pore of the static sparger (m 3 s ±1 ) Fl (=r l 3 áq l 2 ág ±4 áDq ±1 ág ±1 )¯uid number for different forms of bubbles F T inertia force of the displaced liquid (N) H height of the¯uid in the column (m) k L a volumetric oxygen transfer coef®cient (s ±1 ) P/V volumetric power input (N m ±2 s ±1 ) P 0 pressure in the gas bubbles at the surface of the sparged column (N m ±2 ) P B pressure in the gas bubbles formed at the sparger (N m ±2 ) R molar gas constant (R=8.314 kJ K ±1 kmol ±1 ) Re b (=w b ád b áv l ±1 ) Reynolds number of the rising gas bubbles T absolute temperature (K) t p response time of the sensor (s) V M molar volume of ideal gases (V M = 22.41 m 3 kmol ±1 ) w super®cial gas velocity (m s ±1 ) w bbubble rising velocity (m s ±1 ) Dq density difference between gas and liquid (kg m ±3 ) h 90 mixing time for achieving 90% homogeneity (s) q l density of the liquid (kg m ±3 ) gas hold-up in the liquid phase p porosity of the sintered plate g viscosity of the liquid (N m ±2 s) r l surface tension between liquid and gas (N m ±1 ) m l viscosity of the liquid (m 2 s ±1 )

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Section: Discussionmentioning

confidence: 99%

“…[7,15]. The essential advantage of this direct convective gas exchange is the possibility of in¯uencing the oxygen transfer rate into the liquid phase by adjusting the oxygen content in the feed gas.…”

Section: Introductionmentioning

confidence: 99%

Process-engineering characterization of small-scale bubble columns for microbial process development

Weuster‐Botz

Altenbach-Rehm

Hawrylenko³

2001

Bioprocess and Biosystems Engineering

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“…System integrating in situ membrane separation in microbial fermentation process has been shown to have a higher biomass density due to the continuous removal of spent medium and the replenishment with fresh medium (Suzuki 1996;Kamoshita et al 1998a, b). Harmful secreted product, such as ethanol (Ohashi et al 1998) and lactic acid (Kamoshita et al 1998b;Ohashi et al 1999) can be removed in the process and thus enhancing cell viability. It is envisaged that the incorporation of such technology may also benefits plant cell culture in terms of higher yield of metabolite production.…”

Section: Introductionmentioning

confidence: 99%

Improvement of biomass and juvenile hormone III (JH III) production from Cyperus aromaticus cell suspension culture via in situ membrane filtration technology

2010

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The incorporation of membrane filtration technology in plant cell suspension culture was to facilitate in situ medium exchange as well as to eliminate possible introduction of contaminant in the process. The study had been carried out in Cyperus aromaticus cell suspension culture, which is known to produce a bioinsecticide known as juvenile hormone III (JH III). The effect of different volume and flow rate of replenishment toward cell biomass and JH III production were studied. The experimental finding indicates that the flow rate of medium replenishment ranging from 1 to 6 ml min -1 did not affect the cellular structure and morphology the cells. Medium replenishment of 75 v/v% through the membrane system leads to higher cell biomass and metabolite production which correlates to a value as high as 64 and 112%, respectively. The reduction in doubling time, extended exponential growth and increment in specific growth rate were also demonstrated. A 50 v/v% medium replenishment with membrane filtration system was identified to be an economical and feasible application for the production of JH III metabolite at a large scale. The results obtained showed the potential of in situ membrane filtration system for the enhancement of cell biomass and metabolite yield in plant cell culture.

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“…Alternatively, parallel substrate feeding into shake-flasks was achieved with multi-channel syringe pumps feeding up to three shake-flasks [5]. Aerated shake-flasks were designed to overcome the diffusion resistance in the plug and to enhance gas exchange capacity [6,7]. Gas exchange capacity of shakeflasks was improved by rinsing sterile (oxygen-enriched) air through the head space.…”

Section: Introductionmentioning

confidence: 99%

Parallel Bubble Columns with Fed-Batch Technique for Microbial Process Development on a Small Scale

Altenbach-Rehm

Nell

Arnold³

et al. 1999

Chem. Eng. Technol.

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12 small-scale bubble columns, each and every column with a total volume of 500 ml, individually controlled distribution of sterile and humidified air and a pH-probe, were operated in an incubation chamber with temperature control. Intermittent feeding of substrate or inducer, as well as of base for parallel pH-control was achieved by connecting a precision syringe pump to a valve distributor with one 2/2-way miniature valve for each bubble column. This new parallel bio-reactor technique was applied for optimization of the feed profile of the chemical inducer isopropyl-b-D-thiogalactoside (IPTG) to improve the expression of a foreign protein (guanosin-5©-diphosphate-a-D-mannose-pyrophosphorylase, GDP-manPP) under the control of the lac promoter in Escherichia coli. A mean cell density of 8 g l ±1 of recombinant E. coli was achieved in parallel fed-batch fermentations within 9.5 h due to a sufficient oxygen transfer (k L a of up to 0.2 s ±1) and parallel pH-control. The GDP-manPP activity was improved by more than 100 % compared to the standard IPTG pulse within 3 sets of parallel fed-batch fermentations. A Genetic Algorithm was used for optimization of the IPTG feed profile. Scale-up of the optimized fed-batch process into the stirred tank reactor (scale-up factor of 20) was possible, if all reaction conditions were copied exactly with respect to the reactor volume. A final GDP-manPP activity of 715 U l ±1 was measured 3 h after IPTG induction was finished.

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A dense cell culture system for aerobic microorganisms using a shaken ceramic membrane flask with surface aeration

Cited by 12 publications

References 5 publications

Process-engineering characterization of small-scale bubble columns for microbial process development

Process-engineering characterization of small-scale bubble columns for microbial process development

Improvement of biomass and juvenile hormone III (JH III) production from Cyperus aromaticus cell suspension culture via in situ membrane filtration technology

Parallel Bubble Columns with Fed-Batch Technique for Microbial Process Development on a Small Scale

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