1

Simulation of Ethanol Production in Membrane Bioreactor Mohammad Zaid Bin Mohammad Zaini 0637425 Biotechnology Engineering e!artment "ulliyah of Engineering ##$M

Su!er%i&or' (&&oc) Prof) r) (hmed *arig +ameel ,o-Su!er%i&or' (&&i&tant Profe&&or Profe&&or r Maan Ma.an /ahmi 0a&hid (l-1hatib

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ABSTRACT

Membrane bioreactor ha& been the &ub2ect of attention for %alue-added !roduct !roduction& in the&e recent year&) Since the !ea1 of oil !rice in 20063 re&earcher ha& turn their eye& for efficient !roducti%ity of ethanol !roduction) $ntil recently3 little &tudy ha& been made to !redict the micro and heterogeneou& en%ironment of biofilm bioreactor 4a ty!e of membrane bioreactor5 for ethanol !roduction !ur!o&e&) *he !ur!o&e of thi& &tudy i& to &imulate the !roduction of ethanol in biofilm !ac1ed-bed bioreactor by u&ing ,6MS67 Muti!hy&ic&) *he model 8ill co%er the micro and macro en%ironment of !ac1ed-bed bioreactor that ca!able of !redicting the ma&& and reaction di&tribution& along the reactor and 8ithin each biofilm !ellet along the reactor length) *he con&truction of the model 8ill u&e the ad%antage of ,6MS67 &oft8are that are able to &imulate the &!ace-de!endent reacting &y&tem in a heterogeneou& reactor)

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ACKNOWLEDGEMENT

#n the name of (llah3 the Mo&t Merciful and the Mo&t ,om!a&&ionate /ir&tly3 # 8ould li1e to e9!re&& my dee!e&t gratitude to the (lmighty :od3 for ;i& ble&&ing& # 8a& able to com!lete thi& re!ort for Pro2ect < 4B*E 4<=>5 a& a !artial fulfillment of the re?uirement of Bachelor of Biochemical-Biotechnology Engineering &ucce&&fully) # 8ould li1e to ta1e thi& o!!ortunity to 8i&h million of than1& to my &u!er%i&or and co&u!er%i&or3 (&&oc) Prof) r) (hmed *arig +ameel and (&&i&tant Profe&&or Ma.an /ahmi 0a&hid (l-1hatib for their !a&&ion and contribution& in guiding me and hel!ing me in any 8ay that bring u! my !otential in the 8orld of re&earch) *heir indefinite guidance3 &u!!ort3 encouragement and !ro%i&ion in hel!ing me throughout the !ro2ect ha%e been a great contribution and moti%ation for me) /inally3 credit goe& to my familie& and friend& for their %ariou& contribution& and &u!!ort in thi& !ro2ect) *han1 you for your hel!)

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CHAPTER ONE 1.0 Introduction :lobal 8arming i& an i&&ue that ha& been long been debated3 but ha& yet to find it& ultimate &olution) *he i&&ue i& cau&ed by many factor&3 8hich &ome of it i& &lo8ly been dealt 8ith 8hile many are left unattended) *he !artici!ation of "yoto Protocol by more than <60 countrie& from the late&t &tatu& of ratification 4"yoto Protocol3 200553 in that &et concrete target& for de%elo!ed countrie& to reduce the greenhou&e ga&&e& 4:;:5 emi&&ion& that made ma2or contribution to global 8arming3 mar1ed the fir&t action of controlling the addre&&ed i&&ue) @ith more :;: in the mi93 the atmo&!here& 8ill tra! more heat3 acting li1e a thic1ening blan1et) 6ne !romi&ing method to reduce :;: i& by &ub&tituting the current fo&&il fuel 8ith biofuel) Bioethanol and biodie&el are the mo&t commonly !roduced biofuel&) *o date3 they are mainly deri%ed from food cro!& &uch a& corn3 &oybean&3 8heat3 &ugar cane and &orghum) *he&e are referred to a& fir&t generation biofuel&) *he focu& here i& ethanol 8here the de%elo!ment continue& &ince late <>00&) (ccording to ,arolan 4200=53 the rea&on for lac1 of technological ad%ancement of biofuel or bioethanol !roduction i& due to &ocioeconomic force in the&e !a&t decade&) /rom Pena 4200>53 the !roduction in $)S) alone ha& ri&en from le&& than 500 million gallon& in <=>0 to near 5 billion gallon& in 2006) Production in the $nited State& con&i&t& mo&tly of ethanol from corn3 and in BraAil of ethanol from &ugar cane) ,urrently3 mo&t of the ethanol !roduced em!loy& con%entional batch fermentation technology) #n BraAil3 the Melle Boinet !roce&& in 8hich yea&t cell& are &e!arated and reu&ed after acid rin&ing i& indu&triali&ed to almo&t 70B of large-&cale ethanol !roduction) *he&e !roce&&e& are a&&ociated 8ith com!le9 o!eration re?uiring the traditional Cfill and &hutC batch !roce&&3 the danger& of culture contamination3 a general Cin&tabilityC of the &y&tem from an o!eration& &tand!oint3 unreali&ed co&t and !roducti%ity ad%antage& in the !roce&& !lant3 and the fear of genetic drift of the yea&t &train) Moreo%er3 the&e de%ice& ha%e lo8 %olumetric !roducti%itie& and re?uire long fermentation time&) #n addition the high ca!ital and o!erating co&t&3 the continual &tart u! and &hut do8n nature of &uch !roce&&e& ma1e& them difficult to automate and therefore high labour co&t& en&ue) ,on&e?uently3 the u&e of continuou& !roce&&e&3 8hich are &im!le to o!erate3 8ith lo8 energy re?uirement&3 and allo8ing almo&t com!lete utili&ation of the e9!en&i%e &ub&trate&3 8ill

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&ignificantly lo8er the o!erating co&t&) ,a!ital co&t on the other hand3 may be reduced by u&ing mechanically &im!le3 &mall bioreactor& 8ith high rate& of ethanol !roduction) Su!erior ethanol !roducti%itie& may be achie%ed by em!loying a high concentration of yea&t or bacterial cell& 4the cataly&t& for the ethanol !roduction reaction5 8ithin the bioreactor) ;o8e%er3 the !roduction of ethanol u&ing con%entional continuou& !roce&&e& ha& limitation& of maintaining high cell concentration& in the bioreactor) Such limitation& are a&&ociated 8ith lo8 %olumetric !roducti%itie& and long fermentation time& in the&e &y&tem&) Pre&ent trend& in immobili&ed yea&t or bacterial cell bioreactor &y&tem& no8aday& !ro%ide the ethanol indu&try 8ith a method not only for maintaining high cell concentration& in the bioreactor&3 but al&o reducing !roce&&ing time 8ithout &acrificing !roduct ?uality) Production of ethanol ha%e mainly come from a ty!ical fermenter or bioreactor) *he focu& of the &tudy i& to &imulate ethanol !roduction in a !ac1ed-bed biofilm bioreactor)

1.1 Eth no! Ethyl alcohol3 or ethanol3 ,2 ;56;3 i& a clear3 colorle&& li?uid3 8ith a burning ta&te and characteri&tic3 agreeable odor) Ethanol i& the alcohol in &uch be%erage& a& beer3 8ine3 and brandy) Becau&e of it& lo8 freeAing !oint3 it ha& been u&ed a& the fluid in thermometer& for tem!erature& belo8 40D , 440D /53 the freeAing !oint of mercury3 and for other &!ecial lo8tem!erature !ur!o&e3 &uch a& for antifreeAe in automobile radiator&) Ethanol ha& been made &ince ancient time& by the fermentation of &ugar&) (ll be%erage ethanol and more than half of indu&trial ethanol i& &till made by thi& !roce&&) Starch from !otatoe&3 corn3 or other cereal& can be the ra8 material) *he yea&t enAyme3 Ayma&e3 change& the &im!le &ugar& into ethanol and carbon dio9ide) *he fermentation reaction3 re!re&ented by the &im!le e?uation ,6;<266E 2,2 ;56; F 2,62 i& actually %ery com!le9 becau&e im!ure culture& of yea&t !roduce %arying amount& of other &ub&tance&3 including fu&el oil3 glycerin3 and %ariou& organic acid&) *he fermented li?uid3

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containing from 7 to <2 !ercent ethanol3 i& concentrated to =5 !ercent by a &erie& of di&tillation&) #n the !roduction of be%erage& &uch a& 8hi&1ey and brandy3 &ome of the im!uritie&3 8hich &u!!ly the fla%or3 are of great %alue) Much ethanol not intended for drin1ing i& no8 made &ynthetically3 either from acetaldehyde made from acetylene3 or from ethylene made from !etroleum) ( &mall amount i& made from 8ood !ul!) *he biological !roduction of ethanol u&ing yea&t& 8a& an im!ortant commercial !roce&& !rior to <=403 8hen chemical &ynthe&i& from !etrochemical feed&toc1& became more economical) *he recent de!letion of !etroleum re&er%e& ha& re%i%ed an intere&t in fermentation ethanol) #n fact3 8ith the u&e of grain alcohol a& a &ub&titute additi%e for ga&oline3 !roduction reached about 600 million gallon& in <=>63 or about e?ual to the ma9imum !roduction le%el& of the <=30& and <=40&) *he inten&i%e re&earch carried out lately on the con%er&ion of cellulo&ic material& into &ugar&3 follo8ed by biological tran&formation by microorgani&m& into a %ariety of !roduct&3 among them ethanol3 ha& !ro%ided a !otential &ource of chea! ra8 material that may com!ete 8ith !etroleum deri%ati%e&) #n choo&ing a bioreactor ty!e and mode of o!eration3 a biotechnology com!any u&ually balance& medium co&t&3 do8n&tream !roce&&ing co&t&3 technological ad%antage& and &caleability 4;or%ath3 <=>=5) Gariou& factor& ha%e been identified that lead to reduced !roduct co&t& 4Belfort3 <=>=5) /or high %alue biological molecule&3 the co&t of the bioreactor i& relati%ely unim!ortant 8hile other factor& &uch a& increa&ing the !roduct concentration lea%ing the bioreactor can &trongly influence the &elling !rice 4 8yer3 <=>45) *hu&3 relati%ely co&tly de&ign& &uch a& entra!!ed membrane bioreactor& that !roduce !roduct& at e9tremely high e9it concentration& are attracti%e alternati%e& for the&e a!!lication&) /or lo8 %alue biological molecule&3 ho8e%er3 the bioreactor co&t doe& affect the final !roduct !rice &ignificantly) /or the&e a!!lication& con%entional bioreactor& are more a!!ro!riate) ( &ummary of the %ariou& ty!e& of &u&!en&ion and immobiliAed cell bioreactor& i& gi%en in /ig) <)

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/igure <) Summary of different &u&!en&ion and immobiliAed cell reactor& 4%an @eAel3 <=>55

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1." C#!! I$$o%i!i& tion *he u&e of cell immobiliAation ha& been a common laboratory !ractice 8ithin the la&t fe8 decade& a& a method to im!ro%e the !erformance and the economic& of mo&t fermentation !roce&&e&)

1.".1 B 'ic Princi(!#' (lthough immobiliAation techni?ue& differ 8idely3 the common ob2ecti%e can be &ummariAed a& an attem!t to confine the biocataly&t 8ithin the reactor &y&tem3 8hile at the &ame time allo8ing the media containing the &ub&trate and the !roduct& to flo8 in a continuou& manner) *he o%erall effect i& to tran&form a !&eudo-homogeneou& catalytic reaction 4C!&eudoC referring to the &u&!en&ion of the cell& in the media5 into a ty!ical heterogeneou&ly cataly&ed &y&tem) :enerally3 immobili&ed cell &y&tem& can be cla&&ified into four categorie& ba&ed on the !hy&ical mechani&m of cell locali&ation and the nature of the &u!!ort mechani&m&' Hattachment to a &urface3I Hentra!ment 8ithin a !orou& matri93I Hcontainment behind a barrierI and H&elf aggregationI 4"arel3 <=>5J @illaert and Baron <==65 4/ig) <<)<5) /or be%erage 4and other food5 a!!lication&3 &!ecial attention mu&t be !aid to the &election of a!!ro%ed3 food grade com!ound& or to the !re%ention of any lea1age& of unde&ired com!ound& into the be%erage&)

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/igure 2 ,la&&ification of immobili&ed cell &y&tem& according to the !hy&ical locali&ation and the nature of the microen%ironment 4@illaert and Baron <==65)

,ell immobiliAation by ad&or!tion to a &u!!ort material i& a %ery !o!ular method3 becau&e it i& a &im!le3 chea! and fa&t method) Microorgani&m& ad&orb &!ontaneou&ly on a 8ide %ariety of organic and inorganic &u!!ort&) Binding of cell& occur& through interaction& &uch a& Gan der @aal& force&3 ionic bond&3 hydrogen bridge& or co%alent interaction&) Microbial cell& e9hibit a di!olar character and beha%e a& cation& or anion&3 de!ending on the cell ty!e and en%ironmental condition& &uch a& !; of the &olution) /urthermore3 cell !hy&iology ha& a &ignificant influence on the &trength of the adhe&ion) Gariou& rigid &u!!ort material& are a%ailable mainly aimed at a!!lication& in !ac1ed-bed reactor&) iethylaminoethyl 4 E(E5K cellulo&e &u!!ort& ha%e been &ucce&&fully u&ed on indu&trial &cale for the !roduction of alcohol free beer and maturation of green beer 47ommi <==05) #t i& an inert3 non-di&&ol%ing cellulo&e matri93 8hich ha& a non-uniform granular &ha!e) Lea&t cell& are immobili&ed by ionic attraction) ,om!ared to !orou& &u!!ort&3 the bioma&& loading ca!acity of decade&) E(E cellulo&e i& con&iderably lo8er) *able &ummari&e the u&e of immobiliAe cell for fermentation !roce&& in the&e !a&t

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/igure 3) Ba&ic immobili&ed cell &y&tem& 4ada!ted from er%a1o& M @ebb3 Ma&&chelein5)

,ell immobiliAation in !orou& matrice& can be !erformed by t8o different ba&ic method&) #n the fir&t method3 indicated a& gel entra!ment3 the !orou& matri9 i& &ynthe&i&ed in &itu around the cell& to be immobili&ed) #n the &econd method3 cell& are allo8ed to mo%e into a !reformed !orou& matri9) :enerally3 both method& !ro%ide cell !rotection from the fluid &hear and higher cell den&itie& a& com!ared to &urface immobiliAation are reached) ( dra8bac1 of the&e &y&tem& can be ma&& tran&fer limitation&) ;o8e%er3 under&tanding of ma&& tran&fer !henomena 8ithin entra!ment matrice& may allo8 one to &imultaneou&ly !ro%ide different condition& at the carrier &urface and in the interior3 8hich could be attracti%e for coimmobiliAation of different cell ty!e& !erforming con&ecuti%e !roce&&e& 4@illaert3 <===5) *able < i& the recent a!!lication of a biofilm bioreactor)

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*able <) 0ecent (!!lication of Biofilm Bioreactor

6%er the la&t 30 year&3 mo&t of the re&earch concerning the immobiliAation of li%ing microbial cell& 8a& focu&ed on gel entra!ment) Natural !oly&accharide& 4e)g)3 alginate3 chito&an3 !ectate and carrageenan53 &ynthetic !olymer& 4e)g)3 !oly%inylalcohol3 PG(5 and !rotein& 4e)g)3 gelatin3 collagen5 can be gelled into hydro!hilic matrice& under mild condition&3 allo8ing cell

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entra!ment 8ith minimal lo&& of %iability) (& a re&ult3 %ery high bioma&& loading& can be achie%ed) :el& are mo&tly u&ed in the form of &!herical bead& 8ith diameter& ranging from about 0)3 to 5 mm) ;o8e%er3 a di&ad%antage of gel& i& their limited mechanical &tability) #t ha& been fre?uently ob&er%ed that the gel &tructure i& ea&ily de&troyed by the gro8th of the cell& and carbon dio9ide !roduction) Moreo%er3 calcium alginate gel& are 8ea1ened in the !re&ence of !ho&!hate& 48hich i& !re&ent in 8ort5) ;o8e%er3 &e%eral method& ha%e been !ro!o&ed for reinforcement of gel &tructure&) /or e9am!le3 calcium alginate gel can be &trengthened by reaction 8ith !olyethyleneimine3 glutaraldehyde cro&&-lin1ing3 addition of &ilica3 gene!in and !oly%inylalcohol3 or by !artial drying of the gel 4@illaert and Baron3 <==65) /or food !roduction3 a food grade reinforcement method &hould be u&ed) :el or !olymeric matri9 bead& 4u&ually &!herical in &ha!e5 could be further coated 8ith an outer layer to create 4micro5 ca!&ule&) #n tho&e3 the &olid core may al&o be di&&ol%ed 8ithin the ca!&ule to create the li?uid media for cell&) ;o8e%er3 microenca!&ulation i& generally too e9!en&i%e to be u&ed3 thu& only &uited for nongro8ing cell& 40aymond3 20045) (nother a!!roach i& to contain cell& 8ithin a com!artment &e!arated by a !reformed membrane &uch a& hollo8 fibre and flat membrane module&) Entra!ment behind !reformed membrane& re!re&ent& a gentle immobiliAation method &ince no chemical agent& or har&h condition& are em!loyed) $&ually !olymeric microfiltration or ultrafiltration membrane& 8ere u&ed3 although other ty!e& of membrane& 8ere al&o in%e&tigated3 &uch a& ceramic3 &ilicone or ion e9change membrane&) Ma&& tran&fer through the membrane i& de!endent on the !ore &iAe and &tructure a& 8ell a& on the hydro!hobicityOhydro!hilicity and &urface charge) Membrane reactor& !o&&e& &ome in&ight in term of it ad%antage& in &ome &y&tem and di&ad%antageou& for anaerobic !roce&&e&) EnAyme reaction& utiliAing cofactor& and hydroly&i& of macromolecule& are ad%antageou& in membrane reactor&) #mmobiliAation of enAyme& and cell& allo8& reactor& to be o!erated 8ith different hydraulic and biocataly&t retention time&) *he biocataly&t& can be retained either behind a membrane barrier3 in the membrane matri93 or attached to the membrane &urface) *hough ,hang 4<=>75 !ioneered enAyme immobiliAation by microenca!&ulation in <=653 the fir&t true enAyme membrane bioreactor 8a& de%elo!ed by Butter8orth 4<=705 to carry out &tarch hydroly&i& 8ith a-amyla&e) #t ha& been re%ie8ed that for anaerobic culture3 membrane bioreactor 4hollo8 fiber5 !o&&e& &ome di&ad%antage&) /rom Gic1

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0oy 4<=>25 of immobiliAation of 7actobacillu& in the &hell &ide of hollo8 fiber membrane3 the e9!eriment cau&e& the fiber module to be damaged by the gro8ing cell&3 thu& the cell& 8ere able to !enetrate the 8all& and contaminate the recycle %e&&el3 rendering long term o!eration unfea&ible) (l&o3 membrane reactor cau&e& ,62 accumulation a& &aid by Mehaia M ,heryan 4<=>35 and !roblem in the 8a&hout)
*able 2) Selection criteria for cell immobili&ation)

uring the la&t 30 year&3 many different ty!e& of matrice& for immobili&ation ha%e been de%elo!ed3 8hich include& !orou& and non-!orou& !re-formed3 material& &uch a&3 8ood chi!&3 diatomaceou& earth3 %olcanic roc1&3 ceramic&3 &tainle&& &teel3 !orou& bric13 !orou& &intered or &heet gla&&3 !orou& &ilica3 E(E cellulo&e3 PG, chi!&3 !olyurethane cube&3 cotton cloth3 gla&& fibre3 and !lant cell matrice&) Se%eral !olymericmatrice& ha%e been u&ed for cell immobili&ation3 &uch a&3 ,alcium alginate3 1a!!acarrageenan3 !olyacrylamide3 gelatin and e!o9y re&in3 agar3 chito&an3 and &ilica &ol&) *he choice of cell-&u!!orting material for any &!ecific a!!lication ideally &hould meet the &e%eral im!ortant criteria &ummari&ed in *able 3)
*able 3) ,riteria for &election of cell immobili&ation matrice&)

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1."." Ad) nt *#' o+ C#!! I$$o%i!i& tion *he immediate ad%antage& of cell immobiliAation come from !urely chemical reaction conce!t&' higher concentration& of biocataly&t can be achie%ed3 8hich lead to a fa&ter reaction rate3 and the &e!aration of the de&ired !roduct& i& more ea&ily carried out in the ab&ence or at lo8er concentration& of chemically com!le9 &ub&tance&3 &uch a& cell com!onent&) ,ontinuou& o!eration at com!arati%ely high flo8 rate&3 8ithout 8a&hout of the reactor3 i& !o&&ible3 thu& enhancing !roducti%ity) *here i& al&o &ome e%idence to indicate that bound cell &y&tem& generally gi%e higher !roduct yield& and allo8 a better remo%al of to9ic metabolite&3 a& 8ell a& freedom from nutrient de!letion) Performance under heterogeneou& catalytic condition& can be carried out in the !lug flo8 mode3 a& o!!o&ed to 8ell-mi9ed &y&tem&3 8hich in &ome ca&e& 4&uch a& ethanol fermentation3 8here !roduct inhibition may be a factor5 lead& to a more efficient o!eration) ,om!ari&on &tudie& are a%ailable in the literature for batch %er&u& continuou& o!eration and immobiliAed cell reactor& %er&u& continuou& &tirred-tan1 reactor&)

1., Who!# C#!! I$$o%i!i& tion T#chni-u#. Bio+i!$ / Entr ($#nt @hole cell immobiliAation techni?ue& con&i&t of t8o ma2or grou! mention earlier' entra!ment and carrier binding) Entra!ment include& both enclo&ure of a cataly&t behind a membrane and 8ithin a gel &tructure) ,arrier binding include& all method& 8here there i& a direct binding of cell& to 8ater-in&oluble carder& by !hy&ical ad&or!tion or by ionic andOor co%alent bond&) *he term Cbiofilm reactorC i& u&ed here to re!re&ent a grou! of &y&tem& in 8hich gro8th ta1e& !lace on the outer layer of a film of microorgani&m& 8hich i& in direct contact 8ith the &urrounding medium) Potential ma&& tran&fer limitation& are al8ay& !re&ent 8ith entra!ment &y&tem&3 either acro&& the gel matri9 or gel occlu&ion3 or acro&& the &y&tem membrane in membrane reactor&) 6n the other hand3 the carrier binding method allo8& direct contact bet8een the fermentation broth and the biocataly&t3 and the medium flo8 in and out of the &y&tem 8ithout re&triction3 thu&

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minimiAing ma&& tran&fer !roblem&) @hile the &tart-u! of &y&tem& em!loying carrier-binding method& i& influenced by the nature of the &u!!ort and the &!ecific techni?ue em!loyed3 after &ome time of o!eration3 a biocataly&t film i& gro8n and common characteri&tic& to all &y&tem& are found3 thu& allo8ing grou!ing a& &o-called biofilm reactor&)

1.,.1 Bio+i!$ R# ctor' Ad) nt *#' o)#r C#!! Entr ($#nt S0't#$ Biofilm reactor& !re&ent &e%eral ad%antage& o%er entra!!ed &y&tem&) /ir&t3 a& !ointed out by Blac1 4<=>453 it i& highly ad%antageou& to be able to inoculate a fermentor in the normal 8ay and to allo8 cell& to become immobiliAed &im!ly a& a con&e?uence of their gro8th) *hi& com!are& fa%orably 8ith the re?uirement of &!ecial techni?ue& to entra! cell& into matrice& !rior to u&e in the reactor&) Secondly3 gro8th i& a re?uirement for entra!!ed &y&tem&3 due to the lo&& of %iability) of the cell& in the matrice&3 8hich ma1e& reacti%ation 8ith richer media nece&&ary) E%en &o3 &tability of &uch &y&tem& i& generally not good unle&& &ome of the biocataly&t i& re!laced regularly) #n biofilm reactor&3 gro8th i& re?uired and mu&t be !romoted during &tartu!) ;o8e%er3 a& &oon a& a high cell den&ity i& achie%ed3 it i& no longer nece&&ary to !romote cell gro8th3 &o that a &a%ing& in nutrient& can be obtained) #n gel entra!ment &y&tem&3 the mo&t acti%e cell& are at the gel &urface3 &o that agitation of the bead& in &ome in&tance& ha& led to a 50B lo&& of acti%ity due to the lea1age of the outer layer) 6ther di&ad%antage& of cell entra!ment include the re?uirement in &ome ca&e& of a continuou& &u!!ly of chemical& to maintain the hardne&& of the bead& 4&uch a& ,a,#2 for ,a-(lginate matrice&2653 the lac1 of &trength of &ome bead& to &tand hydro&tatic !re&&ure& in !ac1ed bed column&3 and ru!ture of the matrice& due to ,62 !re&&ure)

1.,." Bio+i!$ R# ctor' Ad) nt *#' o)#r Su'(#n'ion Cu!tur# 6ne 8ay to enhance the !roducti%ity of fermentation for %alue-added !roduct !roduction& i& to increa&e the bioma&& in the bioreactor& 4 emirci3 20075) Biofilm reactor& &ho8 many ad%antage& o%er &u&!ended cell reactor&3 e&!ecially in their higher bioma&& den&ity and o!eration &tability) Biofilm reactor& can retain fi%e to ten time& more bioma&& !er unit %olume

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8hen com!ared to &u&!ended reactor&3 8hich contribute& to the increa&e of !roduction rate&3 reduction of 8a&hing out ri&1 8hen conducting a continuou& fermentation at high dilution rate3 and elimination of the need for reinoculation during re!eated-batch fermentation 4/u1uda <==55) (ccording to "unduru and Pometto 4<==6a53 o%erall3 ethanol !roducti%ity in a batch fermentation of P,S biofilm reactor& 8ith Z) mobili& or S) cere%i&iae 8ere three and eight-time& higher than the re&ult& obtained from &u&!en&ion culture&)

1.1 Bio+i!$ R# ctor'. Eth no! Production Biofilm& are defined a& microbial cell layer&3 8hich are irre%er&ibly or re%er&ibly attached on &olid &urface&) *he&e attached cell& are embedded in a &elf-!roduced e9o!oly&accharide matri93 and e9hibit different gro8th and bioacti%ity com!ared 8ith &u&!ended cell&) @ith their high bioma&& den&ity3 &tability3 and !otential for long-term fermentation3 biofilm reactor& are em!loyed for the fermentation and biocon%er&ion3 8hich need large amount of bioma&&) ( ma2or cla&&ification among biofilm reactor& i& ba&ed on the degree of mi9ing' !ac1ed bed& a!!ro9imating !lug flo83 and fluidiAed bed& clo&er to mi9ed flo8) #n ethanol !roduction3 &ome characteri&tic& fa%or one or the other ty!e of o!eration) Sub&trate inhibition fa%or& fluidiAed bed&3 8hile !roduct inhibition ma1e& !ac1ed bed& more !roducti%e) Since ethanol inhibition i& the !redominant of the t8o3 !ac1ed bed& re!re&ent a better o!erating mode and ha%e been the mo&t 8idely u&ed) By a!!ro9imating !lug flo83 the !roducti%ity of !ac1ed bed& i& highly de!endent on the re&idence time in the column) ,hung and Par1 4<=>35 de%elo!ed a fi9ed-bed reactor !ac1ed %ertically 8ith ceramic rod& for continuou& ethanol fermentation by Saccharomyce& cere%i&iae and achie%ed a ma9imum !roducti%ity of 27)5 gP<7P<hP<at gluco&e concentration of<50 gO7 and a dilution rate of <)5 hP<) ( higher ethanol tolerance of the yea&t in the fi9ed-bed biofilm reactor 8a& re!orted 8hen com!ared 8ith the re&ult& in continuou& &tirred-tan1 reactor&)

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#t ha& been noted that a mi9ed culture en%ironment in the reactor &omeho8 !ro%ed to be more efficient in term of !roducti%ity for &ome mi9ture of microorgani&m 8herea& !ro%ed other8i&e for other&) "unduru and Pometto 4<==6a3 b5 a!!lied P,S chi!&3 con&i&ting of !oly!ro!ylene and u! to 25B 48O85 of %ariou& agricultural material& and nutrient&3 a& !ac1edbed reactor& in batch and continuou& ethanol fermentation) *he P,S biofilm reactor& 8ere o!erated u&ing !ure and mi9ed culture& of Z) mobili& or S) cere%i&iae and mi9ed culture& of either of the&e ethanol-!roducing microorgani&m& and the biofilm-forming Stre!tomyce& %irido&!oru& 4*7(5) #n batch fermentation of Z) mobili&3 !ure culture yielded a ma9imum ethanol !roducti%ity of 374 gP< 7P< hP< 444B yield5 a& com!ared to <4> gP< 7P< hP< for mi9ed culture 8ith S) %irido&!oru&) 6n the contrary3 mi9ed culture of S) cere%i&iae and S) %irido&!oru&3 yielded a higher ma9imum !roducti%ity of <=0 gP< 7P< hP< 435B yield5 8hen com!ared to the re&ult& obtained from !ure S) cere%i&iae culture 440 gP< 7P< hP< 3 47B yield5)

/igure 4 (n e9am!le of biofilm reactorJ a5 iagram of the P,S biofilm reactor3 and b5 biofilm of () !ullulan& cell& formed on the P,S &haft)

1.2 P c3#d4B#d R# ctor

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*he !ac1ed bed reactor i& one of the mo&t common reactor& in the chemical indu&try3 for u&e in heterogeneou& catalytic !roce&&e&) #n e&&ence3 the reactor con&i&t& of a container filled 8ith cataly&t !article&) *he&e !article& can be contained 8ithin a &u!!orting &tructure3 li1e tube& or channel&3 or they can be !ac1ed in one &ingle com!artment in the reactor) *he &tructure that i& formatted by the !ac1ed cataly&t !article& ma1e& the modeling of ma&& and energy tran&!ort in the reactor a challenging ta&1) *he difficulty lie& in the de&cri!tion of the !orou& &tructure3 8hich gi%e& tran&!ort of different order& of magnitude& 8ithin the !article& and bet8een the !article&) #n mo&t ca&e&3 the &tructure in bet8een !article& i& de&cribed a& macro!orou& and the !article radiu& can be of the order of magnitude of < mm) @hen a !re&&ure difference i& a!!lied acro&& the bed3 con%ection ari&e in the macro!ore&) *he !ore& in&ide the cataly&t !article& form the micro&tructure of the bed) *he !ore radiu& in the !article& i& often bet8een one and ten micrometer)

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CHAPTER TWO

".1 Pro%!#$ St t#$#nt ,urrently there i& little &tudy on the heteregenou& en%ironment of biofilm !ac1ed-bed bioreactor) #n addition the model that co%er& tho&e a&!ect for ethanol !roduction ha& yet to be con&truct)

"." O%5#cti)#' *he model mu&t &er%e many !ur!o&e&J initially3 it 8ill hel! to determine reaction rate&3 !roduction rate3 gro8th rate and other !arameter& by correlating analytical !rediction& 8ith e9!erimental data) 6nce the !hy&ical con&tant& of the &y&tem are better under&tood3 the model 8ill function to !redict the o!erating beha%ior of current de&ign& under different o!erating condition&) /inally3 the model 8ill be u&ed to o!timiAe future de&ign&) /or the !ac1ed-bed reactor en%ironment3 u&ing a multigeometry a!!roach3 the model !ro%ide& the ma&& and reaction di&tribution& along the reactor and 8ithin each cataly&t !ellet along the reactor length) *hi& ma1e& it !o&&ible to e%aluate the utiliAation of biofilm &tartu!3 o!timal !ellet &iAe3 or inlet tem!erature) /or 2u&tification3 the !ro2ected model 8ill be com!are to an e9i&ting model or re&ult from e9!erimental data) #n addition to that the !ro2ected model 8ill be com!are 8ith other &oft8are3 namely3 Matlab M Ber1eley Madonna)

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CHAPTER THREE

,.0 M#thodo!o*0 *o !redict a !ac1ed-bed biofilm en%ironment3 t8o model& 8ill be con&tructed'
a) Biofilm !ac1ed-bed model u&ing ,6MS67 Multi!hy&ic& 3)5' *o !redict the ma&& and

reaction di&tribution& along the reactor and 8ithin each cataly&t !ellet along the reactor length) b5 0eacting &y&tem model u&ing ,6MS67 0eaction Engineering 7ab <)5' to !redict the &ub&trate con&um!tion rate a& 8ell a& !roduction rate)

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,.1 Bio+i!$ P c3#d4%#d Mod#! u'in* COMSOL Mu!ti(h0'ic' ,.2 ,.1.1 6!o7 Ch rt
Model Definition

Add Geometry

Add Ma !ran "ort Mode

Add PD# Mode

Geometry Modeling

Generate Me $

%om"&ting t$e 'ol&tion

Po t"ro(e in g) *i &ali+ation

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,.1."

Mod#! D#+inition Since ,6MS67 already !ro%ided a modeling e9am!le of a !ac1ed-bed bioreactor3 the

!rocedure that define the model can be &et a& e9am!le3 but it i& &ub2ected to future modification to &ati&fy the biofilm !ellet condition) *he !rocedure done by ,6MS67 i& a& follo8&' <) 2) efining the chemical reaction& e?uation& in%ol%e) efining the all the rate& in%ol%ed) (ccording to Gega M :addy 4<=>>53 8hereby the r9 de&cribe the cell formation rate3 r!3 de&cribe the ethanol formation rate M r& de&cribe the &ub&trate u!ta1e rate&'

8here f4P5 and f '4P5 are the ethanol inhibition relation&hi!&) *he e?uation& need to be modify to &ati&fy the heterogeneou& en%ironment)
3,

efining the rate and ad&or!tion con&tant from (rrheniu& e9!re&&ion&)

8here ( i& acti%ating energy 4+Omol5 and E i& the fre?uency factor&)

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4, ,alculating the !re&&ure dro! in the reactor i& de&cribed by the Ergun e?uation

8here P i& the !re&&ure 41Pa53 Q the !oro&ity3 Dp the !article diameter 4m53 R denote& the ga& %i&co&ity 41gO4mS&553 T the ga& den&ity 41gOm353 and x the reactor length 4m5) u i& the reactor flo8 %elocity 4mO&5 that de!end& on the !re&&ure dro! according to

8here ufeed i& the inlet %elocity and C the total concentration 4molOm35)

5,

efining the ma&& tran&!ort in the reactor i& gi%en by the con%ection and diffu&ion e?uation

8here D i& the diffu&ion coefficient 4m2O&5 and R i& a &ource term 4molO4m3S&55) *he e?uation need& to be &ol%ed for all &!ecie& !artici!ating) 6) ,alculate the 0 from ma&& balance e?uation'

;ere3 D!c i& the effecti%e diffu&ion coefficient in the !article3 c! i& the concentration in the !article3 and R! i& the reaction rate for the heterogeneou& reaction in the !article) #n the biofilm !ore&3 tran&!ort ta1e& !alace by diffu&ion only)

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Becau&e the !ellet i& &!herical the e?uation become&3

8here r 4m5 i& the inde!endent %ariable for the !o&ition along the radiu& of the !article)
7,

efining the boundary condition& of the !article'

8here U denote& the !oro&ity of the !article3 and c and c! re!re&ent the &!ecie& concentration&) *hi& im!lie& &ymmetry at the center of the !article)

,.1., Th# Ph0'ic' S#ttin* *he &ubdomain and boundary condition of e%ery mode add of e%ery &!ecie& in%ol%e in the reaction& ha%e to be in!ut' 1. Subdomain SettingsConvection and Diffusion 2. Boundary ConditionsConvection and Diffusion . Subdomain SettingsPD!" Coefficient #orm $. Boundary ConditionsPD!" %enera& #orm

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,." R# ctin* S0't#$ Mod#! 8'in* COMSOL R# ction En*in##rin* L % 1.2 ,.".1 6!o7 Ch rt *he flo8 chart re!re&ent& the &trategy &ugge&t by ,6MS67 0eaction Engineering 7ab <)5 team to model a chemical reaction &y&tem)

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,."." Th# Mod#!in* Proc#dur# +or Bio+i!$ Bior# ctor S0't#$

Since the &y&tem i& heterogeneou& &y&tem3 thu& it i& a S!ace- e!endent 0eacting Sy&tem&' 8ith Garying *em!erature in *ime) *he modeling !rocedure according to ,6MS67 in the 0eaction Engineering 7ab i&' 1 ) Start ,6MS67 0eaction Engineering 7ab) " ) Select the ty!e of fluid and acti%ate the energy balance) , ) Set u! a ne8 reaction and enter the e9!re&&ion for the reaction de&cribed) 1 ) Set the initial concentration&) 2 ) Set the thermodynamic !ro!ertie&) 9 ) Set the tran&!ort !ro!ertie&) : ) E9!ort to the ,hemical Engineering Module) ; ) Set the boundary condition& and com!ute the &olution in ,6MS67 Multi!hy&ic&) #n order to !erform the e9!ort &te! in the modeling !rocedure3 you need &ome data to calculate the &y&temV& tran&!ort !ro!ertie&' <*he molar 8eight) <*he characteri&tic length of the 7ennard-+one& !otential) <*he energy minimum of the 7ennard-+one& !otential)

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<*he di!ole moment)

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CHAPTER 6O8R E=(#ct#d r#'u!t' *he benchmar1 of the re&ult can be the re&ult from the e9am!le of the !ac1ed-bed reactor model !ro%ided by ,6MS67 team'

/igure 5' *he concentration of reactant& and !roduct& along the reactor length) *he !ellet radiu& rp i& 2)5mm)

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/igure 6' *he concentration of in-!ellet ,6 a& function of reactor !o&ition) *he &caled !ellet radiu& i& gi%en on the r-a9i& and the reactor !o&ition along the 9-a9i&)

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)
/igure >-7=' *he concentration of in-!ellet ,3;6 at three different reactor !o&ition& 453 <53 and 25 cm5) Solid line& re!re&ent& a !ellet radiu& of 2)5 mm and line& 8ith triangular mar1er& re!re&ent& a !ellet radiu& of <)> mm)

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CHAPTER 6I>E Conc!u'ion / R#cco$#nd tion' *hi& !reliminary &tudy for &imulating biofilm !ac1ed-bed bioreactor for ethanol !roduction ha& achie%ed it& !ur!o&e) /rom the !a&t re&earch of cell immobiliAation techni?ue3 it ha& &ho8n that biofilm !ro%e& to be the &uitable &y&tem for anaerobic fermentation) (nd it ha& been &ugge&ted that !ac1ed-bed i& the &uitable method for ethanol3 8hich de&cribe !roduct inhibition) Biofilm !o&e a heterogeneou& catalytic condition& can be carried out in the !lug flo8 mode3 a& o!!o&ed to 8ell-mi9ed &y&tem&3 lead& to a more efficient o!eration) *he fle9ibility of ,6MS67 &oft8are i& ca!able to achie%e the target ob2ecti%e&) Pre%iou& &imulation re&earch ha& !ro%e that ,6MS67 ha& the ca!ability to con&truct com!rehen&i%e model of heterogeneou& catalytic condition& of biofilm !ac1ed-bed bioreactor that co%er the S!ace- e!endent 0eacting Sy&tem&)

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Simulation of Ethanol Production in Membrane Bioreactor

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Simulation of Ethanol Production in Membrane Bioreactor

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