RoumanianBiotechnologicalLetters Copyright2008BucharestUniversity RoumanianSocietyofBiologicalSciences

Vol.13,No.4,2008,pp.38213836 PrintedinRomania.Allrightsreserved ORIGINALPAPER

Comparativeevaluationofradialimpellersefficiencyforbioreactorswith stirredbedofimmobilizedcells 1.Dispersersawtoothandsmithturbine

Receivedforpublication,June1,2008 Accepted,July5,2008
1) 2) ANCAMARCELA LUPTEANU , ANCAIRINA GALACTION , 1) DAN CACAVAL * 1) TechnicalUniversity"Gh.Asachi"ofIasi,FacultyofChemicalEngineeringand EnvironmentalProtection,Dept.ofBiochemicalEngineering,71D.MangeronAvenue, 700050Iasi,Romania,email: dancasca@ch.tuiasi.ro 2) UniversityofMedicineandPharmacyGr.T.PopaofIasi,FacultyofMedical Bioengineering,Dept.ofBiotechnology,913M.KogalniceanuStreet,700454Iasi,Romania, email: galact@from.ro * thecorrespondingauthor

Abstract
Theinfluencesofthemainfactorsonmixingefficiencyanddistributionforabioreactorwith stirred bed of S. cerevisiae immobilized cells in alginate (biocatalysts with 4, 4.6 and 5.2 mm diameters) have been comparativelyanalyzedfor three radialimpellers: disperser sawtooth,Smith turbine and Rushton turbine. The most efficient impeller, from the viewpoint of intensity and uniformityofsuspensioncirculation,wasfoundtobetheSmithturbine.Theoptimumsizeofalginate particleswasof4.6mm,becauseitcompensatesthefrictionspecifictothesmallerparticlesandthe deposition of the bigger particles, the values of mixing time recorded for this size of biocatalysts usingtheSmithturbinebeingthelowestones.

Keywords: bioreactor, stirred bed, immobilized cells, yeasts, mixing, mixing time, radial impeller,Rushtonturbine,dispersersawtooth,Smithturbine.

Introduction
The spectacular applications of the immobilized biocatalysts determined the design and construction of some proper bioreactors, specific or derived from the classical ones. Although these bioreactors are derived from the classical bioreactors and, therefore, their constructiveandfunctionalcharacteristicsarerathersimilarwiththesecondones,theyoffer importantadvantages,namelyas:theincreaseofthethermal,chemicalandtotheshearforces resistanceofthe biocatalysts,the increaseofthenumberoftherepeatedbiosynthesiscycles usingthesameparticlesofbiocatalysts,theeasierrecoveryofthebiocatalystsfromthefinal broths,thediminutionoravoidanceoftheinhibitionprocesses[14]. Thebioreactorsusingimmobilizedbiocatalystcanbedesignedascolumn,stirred,gas lift or membrane bioreactors. They are operated in batch, continuous or semicontinuous systems, with fixed, mobile/stirred, expanded or fluidized bed [5]. Among them, the bioreactorswithstirred/mobilebedofimmobilizedbiocatalystsaresomeofthemoststudied and applied bioreactors, owing to their very similar constructive and operational characteristics to those of the wellknown stirred bioreactors. The main difference between theconstructionsofthetwotypesofbioreactorsconsistsonthepresenceatthebottomofthe formeronesofasievewhichavoidsthebiocatalystsparticleswashout.Themodelsdescribing the flow or the heat and mass transfer in stirred bioreactors, as well as their design and
3821

ANCAMARCELA LUPTEANU, ANCAIRINA GALACTION, DAN CACAVAL

optimizationcanbeeasilyadaptedforthestirredbedbioreactors.But,thesemodelsarevalid onlyforthecontinuousphasefromthebioreactor[5].Duetothedepositiontendencyofthe solidphaseatthebioreactorbottom,totheinternaldiffusionofthesubstrateorproductinto thebiocatalystparticle,themixingand,consequently,theflowofthesesuspensions,aswell asthe mechanismandkineticsoftheprocessesoccurring intothesolidphase become more complexthaninthehomogeneoussystems,thusnewmodelshavingtobeestablishedforthe biocatalystphase[5,6]. Theperformancesofthefermentationprocessesthatarecarriedoutinthebioreactors with stirred bed of biocatalysts are influenced by specific or general factors(the size of the particles[7],geometricalandoperationalcharacteristicsofthevessel [711],concentrationof enzymes/cells into the particles [8,12,13], feed strategy[9,14,15]), among them the mixing efficiencyanditsdistributionbeingthemostimportant[16]. These bioreactors have been used for production of pharmaceuticals [1,17,18], chemicals[19],solventsandbiofuels[20,21],whereasthecurrentstudiesaremainlyfocused onthetreatmentofindustrialormunicipalwastewater [8,9,1214]. Because mixing constitutes one of the main factors controlling these bioreactors performances,being in itsturn influenced by manyconstructiveandoperationalparameters, the analysis and quantification of these influences on mixing efficiency and distribution are requiredforprocessoptimization. One of the most useful criteria for characterization of the mixing intensity is the mixingtime,tm,definedasthetimeneededtoreachagivenmixingintensityatagivenscale, whenstartingfromthecompletelysegregatedsituation[1,23].Thisparameteroffersspecific informationsconcerningthebulkmixinginthesystem(macromixing),respectivelytheflow insidethewholestudiedsystem,butitcannotallowrigorousquantificationofthemesoand micromixing[23].Itcanindicatetheoptimumhydrodynamicregime,thestirrertypethathas tobeused,orcanpredictthemodificationofmixingefficiencyinducedbyscalingup[24,25]. Althoughtheradial impellers,especiallytheRushtonturbine,arewidelyused inthe largescale stirred bioreactors, their applications are limited by the high viscosity and non Newtonian behavior of the broths. Thus, by comparing the information concerning the distribution of circulation intensity, power consumption or shear stress for different double radialstirrers,thefollowingoptimumcombinationsofimpellerswereselectedforsimulated broths: disperser sawtoothand paddle with six blades for water, pitched bladed turbine and Rushtonturbineforbrothswithviscosityupto30cP,pumpermixeranddispersersawtooth forbrothswithhigherviscosity[26]. Bymeansofthepreviousresults,thedevelopmentofacomparativeevaluationofthe mixing induced by different radial impellers for systems containing immobilized cells is required. For selecting the optimum impellers combination, the data on the suspension circulation inside the bioreactors, power consumption and shear effect on biocatalysts particleshavetobetakenintoaccount. Inthiscontext,theaimofourexperimentsistocomparativelystudytheefficiencyof mixing fora bioreactorwithstirred bedof immobilized yeastscells equippedwithdifferent radialimpellers.Thisanalysiswillbemadebymeansofthemixingtimedistribution obtained by vertically changing the position of the pHsensor into the broth, in correlation with the energy consumption. Using the experimental data, the most efficient impeller or impeller combinationwillbeselectedforacertainfermentationbroth. Duetothelargeamountofexperimentaldata,thisstudyisconstitutedinfourparts.In thisone,theresultsobtainedforthedispersersawtoothandSmithturbinearediscussed.

3822

Roum.Biotechnol.Lett.,Vol.13,No.4,38213836(2008)

Comparativeevaluationofradialimpellersefficiencyforbioreactorswithstirredbedofimmobilizedcells 1.Dispersersawtoothandsmithturbine

Materialsandmethod
Theexperimentshavebeencarriedoutin5l(4lworkingvolume,ellipsoidalbottom) laboratory bioreactor (Biostat A, B. Braun Biotech International), with computercontrolled and recorded parameters. The bioreactor characteristics and operating parameters have been presentedinthepreviouspapers[27]. Themixingsystemconsistsofadoublestirrerandthreebaffles.Twotypesofradial impellershavebeenused(Figure1),theexperimentaldatabeingcomparedwiththeprevious onesobtainedfortheRushtonturbine[28].

DisperserSawtooth

SmithTurbine

Figure1. Theradialimpellersusedinexperiments.

The diameter of the two impellers on the shaft, d, was of 64 mm. The inferior impeller has been placed at 64 mm from the bioreactor bottom. The superior impeller was placed on the shaft at a distance of 32 mm from the inferior one, this being the optimum distance form the Ruston turbine, as it was demonstrated in the previous works [28]. The rotation speed was maintained between 500 and 300 rpm, domain that avoids the cave formationatthebrothssurfaceandmechanical disruptionofthebiocatalystsparticles. In the experiments, S. cerevisiae cells immobilized on alginate have been used. The immobilization have been carried out by cells inclusion into the alginate matrix, according withthemethodgiveninliterature[29].Thefollowingdiametersofthebiocatalystspherical particleshavebeenobtained:4,4.6and5.2mm.Thevolumetricfractionoftheimmobilized cellsintothemediavariedbetween7and40%. o The experiments have been carried out at a temperature of 25 C. Any mechanical damageofthebiocatalystduetotheshearforceswasrecordedduringtheexperiments. The mixingefficiency has beenanalyzed by meansofthe mixingtime values,using thetracersmethod[30].Thus,formixingtimedetermination,asolutionof2NKOHhasbeen used as tracer, being recorded the time needed to the media pH to reach the value corresponding to the considered mixing intensity. In this case, the following homogeneity criterionformixing,I,hasbeenconsidered[31]: pH - 0 . 5 DpH I= x 100= 99 % pH where:pH pHvaluecorrespondingtoperfectmixing DpH alloweddeviationfromperfectmixing(DpH=0.02).

Roum.Biotechnol.Lett.,Vol.13,No.4,38213836(2008)

3823

ANCAMARCELA LUPTEANU, ANCAIRINA GALACTION, DAN CACAVAL

The tracer volume was of 0.5 ml, the tracer being injected atthe opposite diametral positiontothepHelectrode(HA405MettlerToledo),at65mmfromthestirrershaftand10 mm from the liquid surface. Because the tracer solution density is close tothe liquid phase density,thetracersolutionflowfollowstheliquidflowstreamsandtherearenoerrorsdueto tracerbuoyancy.ThepHelectrodewasintroducedatfourdifferentpositions,placedvertically from thebioreactorbottomasfollows(Figure2): position1:at20mm position2:at70mm position3:at120mm position4:at170mm. The pH variations were recorded by the bioreactor computerrecorded system and wereanalyzedtocalculate themixingtime.

Resultsanddiscussion
The previous studies on mixing inside of the bioreactors with stirred bed of S. cerevisiae immobilized cells in alginate indicated that the mixing efficiency and its distributionarecontrolledbythesizeandvolumetricfractionofthebiocatalystparticles[28]. In function of the characteristics of biocatalyst particles and operational parameters of the bioreactor,theuniform mixing inthewhole bulkofthesuspensioncould bereached.Thus, for particles with 4 mm diameter and volumetric fraction up to 15%, the optimum rotation speedisof100rpm,increasingto200rpmforparticleswith5.2mmdiameterandthesame domainofsuspensionconcentration.Forbiocatalystswithintermediarysize(particlediameter of 4.6 mm) the uniform circulation of the suspension have been obtained even for more concentrated suspension (of maximum 20% vol.), the optimum rotation speed varying from 150to200rpmwith thevolumetricfractionincreasefrom7to20% [28].

4 3 2 1

Figure2. Theconsideredpositionsinsidethebioreactor.

Theseexperimentsarecarriedoutinthesimilarmannerforanothertwotypesofradial impellers, the disperser sawtooth and the Smith turbine, for selecting the optimum mixing systemforsystemscontainingsuspensionsofimmobilizedyeasts.
3824

Roum.Biotechnol.Lett.,Vol.13,No.4,38213836(2008)

Comparativeevaluationofradialimpellersefficiencyforbioreactorswithstirredbedofimmobilizedcells 1.Dispersersawtoothandsmithturbine

1.Dispersersawtooth FromFigures35importantdifferencescanbeseenbetweenthevariationsofmixing time recorded for the four considered positions inside the bioreactor. Indifferent of the particles size, the dependences can be grouped in two categories for volumetric fraction of biocatalystsupto25%,overthislevelthreetypesofvariationsbeingobserved.
f =0,07
90 80 70 60 50 40 30
40 140

f =0,15
Position1 Position2 Position3 Position4
Position1 Position2 Position3 Position4

t ,s m

t ,s m

120 100 80 60

20 10 0 50 100 150 200 250 300

20 0 50 100 150 200 250 300

Rotationspeed,rpm

Rotationspeed,rpm

f =0,25
220 Position1 Position2 Position3 Position4

f =0,40
300 Position1 Position2 Position3 Position4

t ,s m

t ,s m

200 180 160 140 120

270 240 210 180 150

100

120
80 60 50 100 150 200 250 300

90 50 100 150 200 250 300

Rotationspeed,rpm

Rotationspeed,rpm

Figure3. Theinfluencesofrotationspeedonmixingtimeatdifferentsensorpositionsandbiocatalysts concentrationforthedispersersawtooth(particlediameterof4mm).

Thus, for lower biocatalysts concentration, the shape of the obtained variations is similar fortheinferiorpositions1and2,respectively forthesuperiorones3and4.Forthe inferiorregions,theincreaseofrotationspeedleadstotheinitialreductionofmixingtime,to alevelcorrespondingto150rpm,followedbyitsincreasingintherotationspeeddomainof 150200rpm.Over200rpmthemixingtimeisagainreduced.Thistrendismaintainedalso formoreconcentratedsuspensionsofbiocatalysts,buttheminimumandmaximumofmixing time become less evident. Moreover, by increasing the volumetric fraction of alginate particles, the rotation speed which correspond to the two extremes are moved to higher values. Consequently, for 40% vol. immobilized cells the maximum level of mixing time cannotbereachedfortheexperimentalrotationspeeddomain.

Roum.Biotechnol.Lett.,Vol.13,No.4,38213836(2008)

3825

ANCAMARCELA LUPTEANU, ANCAIRINA GALACTION, DAN CACAVAL f =0,07

120 Position1 Position2 Position3 Position4
180 t ,s m 150 120 90 60 30 0 Position1 Position2 Position3 Position4

f =0,15

t ,s m

100 80 60 40 20 0 50 100 150 200

250

300

50

100

150

200

250

300

Rotationspeed,rpm

Rotationspeed,rpm

f =0,25
t ,s m
270 Position1 Position2 Position3 Position4

f =0,40
320 280 240 Position1 Position2 Position3 Position4

t ,s m

240 210 180 150 120 90

200 160 120

60 50 100 150 200 250 300

50

100

150

200

250

300

Rotationspeed,rpm

Rotationspeed,rpm

Figure4. Theinfluencesofrotationspeedonmixingtimeatdifferentsensorpositionsandbiocatalysts concentrationforthedispersersawtooth(particlediameterof4.6mm).

For biocatalysts concentration below 15% vol., the intensification of rotation speed continuouslyimprovesthesuspensioncirculationatthebioreactortop.Theincreaseofthesolid phaseamountleadstothestrongerdifferentiationofthemixingtimevariationforthepositions3 and4.Therefore,the shapeoftherecorded curves forposition3remains similar fortheentire domain of alginate particles concentration. But, for the superior position 4 the obtained results indicate a minimum level for mixing time, this parameter increasing then. The rotation speed which corresponds to the maximum efficiency of mixing is displaced to lower values by concentrating the suspension. Consequently, for biocatalysts volumetric fraction over 40% and rotationspeedover200rpmthemixingtimeforposition4isratherequalwiththatforposition3, becominghigherforthealginateparticleswithdiameterof4.6mm.
f =0,07
140 Position1 Position2 Position3 Position4
200 t ,s m

f =0,15
Position1 Position2 Position3 Position4

t ,s m

120 100 80 60 40

160

120

80

40

20 50 100 150 200 250 300

50 100 150 200 250 300

Rotationspeed,rpm

Rotationspeed,rpm

3826

Roum.Biotechnol.Lett.,Vol.13,No.4,38213836(2008)

Comparativeevaluationofradialimpellersefficiencyforbioreactorswithstirredbedofimmobilizedcells 1.Dispersersawtoothandsmithturbine f =0,25

300 Position1 Position2 Position3 Position4 360

f =0,40
Position1 Position2 Position3 Position4

t ,s m

270 240 210

t ,s m

320 280 240

180 200 150 120 90 50 100 150 200 250 300 160 120 50 100 150 200 250 300

Rotationspeed,rpm

Rotationspeed,rpm

Figure5. Theinfluencesofrotationspeedonmixingtimeatdifferentsensorpositionsandbiocatalysts concentrationforthedispersersawtooth(particlediameterof5.2mm).

The above presented data differ significantly from the previous ones obtained for simulated fermentation broths without solid phase [26]. The presence of the solid phase modifies the system behavior, owing to the appearance of a supplementary phenomenon, namely its deposition. In this case, the mixing has to avoid the deposition tendency of the biocatalystsandtouniformly dispersethebrothscomponents. Thehighestconcentrationofsolidphaseisintheinferiorregionofthebioreactor.For this reason, the biggest values of mixing time have been recorded for positions 1 and 2, respectively.Theparticularvariationofmixingtimecorrespondingtotheinferiorregioncan betheresultoftheinterferenceofthestreamsinducedbytheimpellersplacedat0.5donthe shaft, phenomenon that is amplified by the bottom effect and solid phase collision. Therefore,the hindrance of suspension circulation is more importantthan for the simulated brothswithoutsolidphase.Theincreaseoftherotationspeedoverthelevelneededtoreach the maximum mixing time diminishes these negative effects, thus leading to the intensification of the suspension circulation in this region. The accumulation of solid phase diminishestheinfluenceoftherotationspeedand,consequently,themagnitudeoftheabove discussedphenomenaisreducedathigherbiocatalystsconcentration. Althoughthepositions3and4areplacedatahigherdistancefromtheimpellersand, therefore, from the region in which the turbulence is generated, the mixing time in these regions is lower than that for the inferior positions, due to the lower concentration of biocatalysts. The influence of the rotation speed in the superior region is reduced by the increase of the solid phase amount, due to its dispersion in the whole bulk volume of the media with the mixing intensification. But, for particles concentration over 15% vol., the dependence between the mixing time and rotation speed differs from position 3 to 4. According to the above discussed results, the particular variation recorded for position 4 is due especially to the deposition of solid phase and less to the friction forces between the alginate particles, effect promoted by the low pumping capacity of the disperser sawtooth. The increase of particles size from 4 to 5.2 mm amplifies the magnitude of the variations obtained for positions 1 and 2 and attenuates those recorded for positions 3 and 4, this demonstratingtheaboveconclusion. Byanalyzingcomparativelythe influenceofthe rotationspeedon mixingefficiency anditsdistributioninsuspensionsofimmobilizedS.cerevisiaeinalginateforadoublestirrer

Roum.Biotechnol.Lett.,Vol.13,No.4,38213836(2008)

3827

ANCAMARCELA LUPTEANU, ANCAIRINA GALACTION, DAN CACAVAL

ofRushtonturbinetype[28]andoneofdispersersawtoothtype,importantdifferenceshave beenobserved,asfollows: Positions 1 and 2: indifferent of the size and concentration of the biocatalysts particles, the Rushton turbine is more efficient for the entire rotation speed domain used, owing to the lower dispersing capacity and to the more pronounced streams interferenceofthedispersersawtooth. Position 3: the disperser sawtooth induces a more intense mixing for biocatalysts volumetric fractionbelow15%,butonly forrotationspeed valuesdirectlyrelatedto the alginate particles size and concentration. Thus, for biocatalysts particles with 4 mmdiameterand7% vol.concentration,thedispersersawtooth is moreefficient for theentiredomainofrotationspeed,andonlyforrotationspeedover100rpmformore concentratedsuspensions.Forparticleswithdiameterof4.6 mm,this stirrer is more efficientonlyforrotationspeedhigherthan150rpmforbiocatalystsconcentrationup to 7% vol., and only for rotation speed over 200 rpm for higher concentration of biocatalysts. In the case of the biggest particles, the disperser sawtooth generates a more intense circulation only for rotation speed over 200 rpm, if the suspension concentration is below 15% vol. For solid phase concentration over 15% vol., the Rushton turbinebecomesmoreefficientnotdependingon particlessize. Position4:inallcases,thedispersersawtoothpromotesthemostintensecirculation. But,theseconclusionsshouldbeprudentlyanalyzed,becausethemixingpromotedby the disperser sawtoothfor the superior positions is apparently more intense, due to the low amountofsolidphasedispersedinthesuperiorregion. TheFigure6doesntindicateanypossibilitytoreachanuniformmixinginthewhole bulk volume of the suspension if the disperser sawtooth is used. Contrary, the uniform dispersion of the solid phase can be obtained for the Rushton turbine at optimum rotation speedvaluesof100200rpm,butonlyforbiocatalystsconcentrationbelow15%vol.[28]. The increase of the biocatalysts particles size exhibits a negative influence on the mixing time. Therefore, for the disperser sawtooth it can be concluded that the solid phase depositioncontrolstheefficiencyofmixing. 2.Smithturbine This type of impeller disperses the gases better than the Rushton turbine, being recommendedforaerobicfermentationprocesses. As in the case of disperser sawtooth, the correlations between the mixing time and rotation speed are of two types, due to the deposition tendency of biocatalysts. The differencesbetweenthevariationscorrespondingtothefourpositionsarediminishedwiththe increaseofthesolidphaseconcentration(Figures79). For lower volumetric fraction of biocatalysts, the mixing time initially decreases by acceleratingtherotationspeedto100rpm, increases forrotationspeed varying from100to 150rpm,decreasingstronglythen.Thisvariationhasbeenrecordedforpositions1,2and3, butitsamplitudediffersfromonepositiontoanother.Therefore,thehighestvalueofmixing time, corresponding to 150 rpm, is reached for position 2, as the result of the streams interference and solid phase collision with the baffles or bioreactor wall. The relative importance of these phenomena is maximum at 150 rpm, for the rest of the rotation speed domain the mixing intensity for position 2 being closer or superior to that obtained for position1.

3828

Roum.Biotechnol.Lett.,Vol.13,No.4,38213836(2008)

Comparativeevaluationofradialimpellersefficiencyforbioreactorswithstirredbedofimmobilizedcells 1.Dispersersawtoothandsmithturbine f =0,07

80
50rpm 100rpm 150rpm 200rpm 250rpm 300rpm

dP =4mm
t ,s m
270 240 210 180 150 120

f =0,40
50rpm 100rpm 150rpm 200rpm 250rpm 300rpm

t ,s m

70 60 50 40 30 20 10 0 1 2 3

Position

Position

f =0,07
110
50rpm 100rpm 150rpm 200rpm 250rpm 300rpm

dP =4.6 mm
320

f =0,40

t ,s m

t ,s m

100 90 80 70 60 50 40 30 20 10 1 2 3

280

240

50rpm 100rpm 150rpm 200rpm 250rpm 300rpm

200

160

120

Position

Position

f =0,07
t ,s m
140 120 100 80 60 40 20 1 2 3 4
50rpm 100rpm 150rpm 200rpm 250rpm 300rpm

dP =5.2 mm
t ,s m
360 330 300 270 240 210 180 150 120

f =0,40
50rpm 100rpm 150rpm 200rpm 250rpm 300rpm

Position

Position

Figure6. Variationofmixingtimewiththepositioninsidethebiocatalystssuspensionforthedisperser sawtooth.

Roum.Biotechnol.Lett.,Vol.13,No.4,38213836(2008)

3829

ANCAMARCELA LUPTEANU, ANCAIRINA GALACTION, DAN CACAVAL

f =0,07
40 Position1 Position2 Position3 Position4 50

f =0,15
Position1 Position2 Position3 Position4

t ,s m

t ,s m
40 30 20 10 50 100 150 200

30

20

10

50

100

150

200

250

300

250

300

Rotationspeed,rpm

Rotationspeed,rpm

f =0,25
t ,s m
90 Position1 Position2 Position3 Position4
120

f =0,40
Position1 Position2 Position3 Position4

t ,s m

80

110 100 90 80

70

60

70 60 50

50

40 50 100 150 200 250 300

40 50 100 150 200 250 300

Rotationspeed,rpm

Rotationspeed,rpm

Figure7. Theinfluencesofrotationspeedonmixingtimeatdifferentsensorpositionsandbiocatalysts concentrationforthe Smithturbine (particlediameterof4mm).

The variation of mixing time for position 3 is similar, but the influence of the mentioned phenomena is significantly attenuated, owing to the distance from the bottom regionwithconcentrateddepositofsolidphase.Byintensifyingtheagitation,thebiocatalysts are dispersed also in the superior region, and, consequently, their volumetric fraction increasesinregion3.For rotationspeedhigherthan200rpm,thisprocess,cumulatedwiththe lower turbulence transmitted in position 3, leads to the increase of mixing time over the valuesrecordedforpositions1and2.
f =0,07
Position1 Position2 Position3 Position4
40

f =0,15
Position1 Position2 Position3 Position4

30

t ,s m

t ,s m
30 20 10 50 100 150 200

20

10

0 50 100 150 200 250 300

250

300

Rotationspeed,rpm

Rotationspeed,rpm

3830

Roum.Biotechnol.Lett.,Vol.13,No.4,38213836(2008)

Comparativeevaluationofradialimpellersefficiencyforbioreactorswithstirredbedofimmobilizedcells 1.Dispersersawtoothandsmithturbine

f =0,25
t ,s m
80 Position1 Position2 Position3 Position4
110 100 90 80 70

f =0,40

70

60

Position1 Position2 Position3 Position4

50
60 50 40

40

30 50 100 150 200 250 300

t ,s m

30 50 100 150 200 250 300

Rotationspeed,rpm

Rotationspeed,rpm

Figure8. Theinfluencesofrotationspeedonmixingtimeatdifferentsensorpositionsandbiocatalysts concentrationforthe Smithturbine (particlediameterof4.6mm).

Themixingfortheposition4iscontinuouslyimprovedbyrotationspeedincreasing, butitbecomeslessefficientcomparedwiththepositions1and2forrotationsspeedover200 rpm,duetothedispersionofthesolidphasealsointhesuperiorregion.Althoughtheposition 4isplacedatthehighestdistancefromthesuperiorimpeller,themixingtimefortheposition 4 was inferior tothat related tothe position 3 for the entire domain of rotation speed. This resultcanbeattributedeithertothelowerconcentrationofbiocatalystsinthisregion,andto thepropagation intheposition3ofthe negative effectof streams interferenceandparticles collision.
f =0,07
40 Position1 Position2 Position3 Position4 50

f =0,15
Position1 Position2 Position3 Position4

t ,s m

t ,s m
40 30 20

30

20

10 50 100 150 200 250 300

50

100

150

200

250

300

Rotationspeed,rpm

Rotationspeed,rpm

Roum.Biotechnol.Lett.,Vol.13,No.4,38213836(2008)

3831

ANCAMARCELA LUPTEANU, ANCAIRINA GALACTION, DAN CACAVAL f =0,25

t ,s m
100 90 80 70 60 50 40 50 100 150 200 250 300 Position1 Position2 Position3 Position4 130

f =0,40
Position1 Position2 Position3 Position4

t ,s m

120 110 100 90 80 70 60 50 50 100 150 200

250

300

Rotationspeed,rpm

Rotationspeed,rpm

Figure9. Theinfluencesofrotationspeedonmixingtimeatdifferentsensorpositionsandbiocatalysts concentrationfortheSmithturbine(particlediameterof5.2mm).

Thesevariationsaremaintainedforbiocatalystsconcentrationupto15%vol.But,the rotation speed corresponding to the maximum mixing time for the positions 1, 2 and 3 is moved to higher values with the particles concentration increase, becoming of 200 rpm. Moreover,thesuspensioncirculationintheposition3ismoreintensethanthatintheposition 1 or 2 for a larger domain of rotation speed, this underlining that the alginate particles depositionandcollision,aswellasthestreamsinterference,significantlyreducetherelative efficiencyofthemixingintheinferiorregionofthebioreactorforconcentratedsuspensions of biocatalysts. For the same reason, the mixing time obtained for position 4 continuously decreases with rotation speed acceleration and reaches the lower values comparatively with theotherthreepositions. Forbiocatalystsvolumetric fractionover15%,theshapeofthe variationsdescribing the rotation speed influence on the mixing time in the considered regions inside the bioreactors become closer, the intensity of the suspension circulation increasing with the acceleration of rotation speed. The lowest values of mixing time have been recorded for position 4. On the other hand, for rotation speed below 200 rpm the highest mixing time values have been obtained for position 1, over this level of rotation speed for position 2. These results, as well as the comparison with the mixing induced by Smith turbine in simulatedfermentationbrothswithoutsolidphase[26],suggestthatthecirculationvelocityof the suspension is reduced by the alginate particles collision with the baffles, phenomenon amplifiedinmoreconcentratedsuspensions. Althoughthevariationsofmixingtimeforthefourpositionsaresimilar,therelative importance of the friction between the alginate particles and of their deposition can induce modification of these variations shapes. Thus, due to the equilibrium existing between the friction forces, specific to smaller particles, and the deposition to the bioreactor bottom, specific to the bigger ones, the minimum and maximum of mixing time are attenuated for particleswiththeintermediarydiameterof4.6mm.Inthecaseofalginateparticleswith5.2 mmdiameter,thepronouncedtendencyofparticlesdepositiontothebioreactorbottomleads totheamplificationoftheseextremespointsandtothemoreevidentdifferentiationbetween thepositions1and2comparedwithpositions3and4.Forthesmallestbiocatalysts(diameter of 4 mm), the deposition tendency is considerably diminished, therefore the variation recordedforposition3israthersimilartothosecorrespondingtopositions1and2. The comparative analysis of the mixing promoted by Smith and Rushton turbines indicatesthattheformeroneoffershigherefficiencyinthemostofthestudiedcases:
3832

Roum.Biotechnol.Lett.,Vol.13,No.4,38213836(2008)

Comparativeevaluationofradialimpellersefficiencyforbioreactorswithstirredbedofimmobilizedcells 1.Dispersersawtoothandsmithturbine

Position1:indifferentofthebiocatalystssize,theSmithturbineismoreefficientfor particlesconcentrationbelow25%vol.Formoreconcentratedsuspensions,theSmith turbine induces a more intense circulation only for biocatalysts with 4 and 5.2 mm diametersandrotationspeedupto150200rpm,thenthemixingefficiencybecoming similartothatofRushtonturbine.But,forthebiocatalystswithintermediarysize(4.6 mm), the Rushton turbine is recommended for rotation speed over 100 rpm and volumetricfractionhigherthan25%. Position2:inthecaseofbiocatalystswithdiameterof4mm,theSmithturbineoffers a more efficient mixing for the entire considered domain of rotation speed of solid phaseconcentration.Forthebiggestalginateparticles,thisimpellerinducesanintense mixingonlyforsolidphaseconcentrationover25%vol.androtationspeedbelow150 rpm, the mixing efficiency being rather similar to that promoted by the Rushton turbineintherest.But,theefficiencyoftheSmithturbineforparticleswithdiameter of4.6mmisthesameasintheposition1. Positions3and4:inallcases,theSmithturbinepromotesthemostefficientmixing. Contrarytothedispersersawtooth,theuseoftheSmithturbineoffersthepossibility toreach the uniform circulation into the whole bulk volume of the suspension for a certain domain of particles concentration and rotation speed values, both correlated with the biocatalysts size, as in the case of Rushton turbine (Figure 10). Therefore, for biocatalysts withdiametersof4and4.6mm,theuniformmixingisobtainedonlyforvolumetricfraction upto15%,at100and200rpmforconcentrationof7%vol.,respectivelyat250rpmformore concentratedsuspensions.Theuniformdispersionofthesolidphase isreached forparticles with5.2mmdiameteronlyforconcentrationbelow7%at250rpm. Becausethebiocatalystwithintermediarydiameterof4.6mmdiminishesthefriction forces between the particles compared with the smaller biocatalysts, and the deposition tendencycomparedwiththebiggerones,thelowestvaluesofmixingtimehavebeenrecorded forthesesizeof alginateparticles(Figure11).

Conclusions
Byanalyzingcomparativelythemixingintensityanditsdistributionintoabioreactor with stirred bed of yeast cells immobilized in alginate (particles with 4, 4.6 and 5.2 mm diameters) using three radial impellers (disperser sawtooth, Smith turbine vs. Rushton turbine),thefollowingconclusionscanbedrawn: 1. The less efficient impeller was the disperser sawtooth, especially due to the low pumping capacity which cannot avoid the solid phase deposition at the bioreactor bottom. Therefore, the increase of the biocatalysts size led to the significant reduce of the mixing efficiency. 2. The Smith turbine offers the most efficient mixing for a large domain of biocatalystsconcentrationandrotationspeed.Itcanalsoinduceanuniformcirculationofthe suspensionforcertainvaluesofrotationspeedandbiocatalystsvolumetricfractionupto15%, similartotheRushtonturbine. The most efficient mixing has been obtained for biocatalyst particles with 4.6 mm diameter, due to the equilibrium existing between the friction forces, specific to smaller particles,anddepositionto thebioreactorbottom,specifictothebiggerones. These studies will be developed for other radial impellers that could be used for bioreactorswithstirredbedofimmobilizedcells. Roum.Biotechnol.Lett.,Vol.13,No.4,38213836(2008)
3833

ANCAMARCELA LUPTEANU, ANCAIRINA GALACTION, DAN CACAVAL

f =0,07

dP =4mm
50rpm 100rpm 150rpm 200rpm 250rpm 300rpm

f =0,40
50rpm 100rpm 150rpm 200rpm 250rpm 300rpm

120 t ,s m 110 100 90 80

40 t ,s m 30 20

70 60
10

50
0 1 2 3 Position 4

40 1 2 3 Position 4

f =0,07
30 25 20 15 10 5

dP =4.6 mm
50rpm 100rpm 150rpm 200rpm 250rpm 300rpm

f =0,40
50rpm 100rpm 150rpm 200rpm 250rpm 300rpm

110 t ,s m 100 90 80 70 60 50 40

t ,s m

0 1 2 3 Position 4

Position

f =0,07
40 35 30 25 20 15 10 1 2 3 Position 4

dP =5.2 mm
50rpm 100rpm 150rpm 200rpm 250rpm 300rpm 130 t ,s m 120 110 100 90 80 70 60 50 40 1 2

f =0,40
50rpm 100rpm 150rpm 200rpm 250rpm 300rpm

t ,s m

Position

Figure10.VariationofmixingtimewiththepositioninsidethebiocatalystssuspensionfortheSmithturbine.

3834

Roum.Biotechnol.Lett.,Vol.13,No.4,38213836(2008)

Comparativeevaluationofradialimpellersefficiencyforbioreactorswithstirredbedofimmobilizedcells 1.Dispersersawtoothandsmithturbine

f =0,07
t ,s m 35 30 25 100rpm 200rpm 300rpm

Position1
110 t ,s m 100 90

f =0,40
100rpm 200rpm 300rpm

20 15 10 5 60 0 4.0 4.4 4.8 dP,mm 5.2 4.0 4.4 4.8 d ,mm P 5.2 80 70

f =0,07
35 t ,s m 30 25 20 15 10 100rpm 200rpm 300rpm

Position2
100 t ,s m

f =0,40
100rpm 200rpm 300rpm

90

80

70 5 0 4.0 4.4 4.8 dP,mm 5.2 60 4.0 4.4 4.8 d ,mm P 5.2

f =0,07
30 t ,s m 100rpm 200rpm 300rpm

Position3
90

f =0,40
100rpm 200rpm 300rpm

25

t ,s m
80 70 60 50 40

20

15

10

5 4.0 4.4 4.8 d ,mm P 5.2

4.0

4.4

4.8

d ,mm P

5.2

f =0,07
25 t ,s m 100rpm 200rpm 300rpm

Position4
75 t ,s m 70 65 60

f =0,40
100rpm 200rpm 300rpm

20

15

55 50

10

45 40

5 4.0 4.4 4.8 dP,m m 5.2

35 4.0 4.4 4.8 d ,mm 5.2 P

Figure11.Variationofmixingtimewiththe alginateparticlesdiameterfortheSmithturbine.

Roum.Biotechnol.Lett.,Vol.13,No.4,38213836(2008)

3835

ANCAMARCELA LUPTEANU, ANCAIRINA GALACTION, DAN CACAVAL

Notations d impellerdiameter,mm dP biocatalystparticlediameter,mm tm mixingtime,s f biocatalystsvolumetricfraction,%

This work was included in the Grant PNCDI II 21048/2007 supported by The National CentreforProgramsManagement(CNMP)

References
1. D. CACAVAL, C. ONISCU, A.I. GALACTION, Inginerie biochimic i biotehnologie. 2. Bioreactoare,InterGlobal,Iai,2002,pp.15 17. 2. R.H.WIJFFELS,R.M.BUITELAAR,C.BUCKE,J.TRAMPER, Immobilized Cells: Basics & Applications.ProgressinBiotechnology,vol.11,EditorialElsevier,Amsterdam,1996. 3. W. TISHER, F. WEDEKIND, H.D. FESSNER, Immobilized Enzymes: Methods and Applications.Biocatalysis.TopicsinCurrentChemistry,95126(1999). 4. M.B. ANGELOVA, S.B. PASHOVA, C. SLOKOSKA, EnzymeMicrob.Technol., 26, 544549(2000). 5. LUPTEANU A.M.,GALACTION A.I.,CACAVAL D., Roum. Biotechnol. Lett., 12, 3131 3138(2007). 6. W. HARTMEIER,Immobilizedbiocatalysts,SpringerVerlag,Berlin,1988. 7. DAGUER.R., HABBENC.E., PIDAPARTIS.R.,WaterSci.Technol., 26,24292432(1992). 8. SUNG S., DAGUER.R.,WaterEnviron.Res., 67,294301(1995). 9. ANGENENT L.T., DAGUE R.R., 50th Purdue Industrial Waste Conference Proceedings, AnnArborPress,Chelsea,Michigan,365377(1995). 10. ZHANG R., YIN Y., SUNG S., DAGUE R.R., 51st Purdue Industrial Waste Conference Proceedings,AnnArborPress,Chelsea,Michigan,315320,(1996). 11. BRITOA.G., RODRIGUESA.C., MELOF.L.,WaterSci.Technol.,35,93198(1997). 12. FERNANDESL., KENNEDYK.J., NING Z., WaterRes., 27,16191628(1993). 13. TIMURH., STURK I.,WaterRes., 33,32253230(1999). 14. BAGLEYM., BRODKORB T.S.,WaterEnviron.Res., 71,13201332(1999). 15. RATUSZNEI S.M., DOMINGUESRODRIGUES J.A., MORALESDE CAMARGO E.F., RIBEIRO R., ZAIAT M.,BioresourceTechnol.,87,203209(2003). 16. T. GU, M.J. SYU, Biotechnol.Prog.,20,14601466(2004). 17. G.B. BORGLUM, J.J. MARSHALL,Appl.Biochem.Biotechnol.,9,117130(1984). 18. TANQ., SONG Q., WEID.,EnzymeMicrob. Technol., 39,11661172 (2006). 19. KENNEDY J.F.,CABRAL J.M.S., in Applied biochemistry and bioengineering (I. Chibata, L.B.Wingardeds.),vol.4,AcademicPress,NewYork,1983,pp.53151. 20. LINKOP., LINKOY.Y.,Crit.Rev.Biotechnol., 1,289338(1984). 21. LUONG J.H.T., TSENG M.C., Appl.Microbiol.Biotechnol.,19,207216(1984). 22. W.W. XI, J.H. XU, ProcessBiochem.,40, 21612166(2005). 23. BUJALSKIJ.M,Ph.D.Thesis,UniversityofBirmingham,2003,p.22. 24. KRAMERSH., BAARSM., KNOLLW.H.,Chem.Eng.Sci.,2,3542(1953). 25. NIENOWA.W.,Chem.Eng.Sci.,52,25572264(1997). 26.CACAVAL D., GALACTION A.I., FOLESCU E., Chem.Ind.Chem.Eng. Quart., 13,119(2007). 27.CACAVAL D., GALACTION A.I., TURNEA M., J.Ind.Biotechnol.Microbiol., 34,3547(2007). 28.GALACTION A.I., LUPTEANU A.M., CACAVAL D., Environ. Eng. Manag. J., 6, 101110 (2007). 29. WILLIAMSD., MUNECKED. M., Biotechnol.Bioeng., 23,18131825 (1981). 30.ONISCU C., GALACTION A.I., CACAVAL D., UNGUREANU F., Biochem.Eng.J., 12,6169(2002). 31.VANT RIET K.,TRAMPER J.,BasicBioreactorDesign,M.DekkerInc.,NewYork,1991, pp.183.
3836

Roum.Biotechnol.Lett.,Vol.13,No.4,38213836(2008)