This action might not be possible to undo. Are you sure you want to continue?

**Professor Dhinakar S. Kompala Department of Chemical and Biological Engineering ni!ersit" of Colorado Boulder# Colorado $%&%'
**

(ith Pol"math programs written b" ).S. *ogler and Som Ghosh

Modifications to Monod Model

The empirically based Monod growth rate equation

has become popular, compared to the other proposed rate equations for cell growth, due to its similarity with the mechanistically derived

Michaelis-Menten rate law for enzyme-catalyzed reaction rate. The Monod equation is capable of explaining or simulating the exponential growth phase followed by the decelerating growth phase in the cell concentration during batch growth dynamics, when coupled with dynamic balance equation for the substrate concentration:

where YC/S is the stoichiometric yield coefficient of grams of cell mass produced from a gram of substrate.

igure !. Typical growth phases in batch cultures of bacterial cells

The initial lag and accelerating growth phases which are not simulated by the Monod growth rate equation can be easily simulated by ma"ing a slight modification to the original Monod equation given above. This modification invo"es a rate-limiting enzyme involved in the growth processes that may not be present at sufficient levels in the inoculum or starting culture. #hile cell growth is a complex process mediated by thousands of enzymes, it may be sufficient to hypothesize $ust a single enzyme that may be rate-limiting during the initial lag growth phase for the purpose of simulating the lag.

%ncorporating the effect of varying "ey enzyme concentration into the Monod growth "inetics, we can write

&'.(.).*+ where ER represents the relative amount of the "ey enzyme in the cell. This modified Monod rate equation follows the Michaelis-Menten rate equation closely, which is more correctly written as

&'.(.).)+ where ET represents the total enzyme concentration in the reaction mixture.

'elative "ey enzyme content inside the cells, ER, may be written as , where e is the intracellular content of the "ey enzyme, with the units of g enzyme,g cell mass, and emax is is the maximum enzyme concentration in the cell, also with units of g enzyme,g cell mass. The balance equation for the intracellular enzyme content can be written in terms of eCc, which has the units of g enzyme,culture volume, as

&'.(.).-+ where a and b are enzyme synthesis and degradation rate constants. .sing the product rule, we can expand the above equation to write the balance equation for e as:

&'.(.)./+ The last term in the above balance equation for the intracellular enzyme is the dilution term due to cell growth, which will be obtained similarly for all intracellular species. This equation can be solved to produce the curve added at the bottom of the batch growth dynamics as shown in igure 0. The maximum level of the intracellular enzyme emax can be determined easily by setting the above equation to zero and solving for its steady state value. %n terms of the model parameters, the maximum level of intracellular enzyme obtained during the exponential growth phase &when Cs is much greater than Ks+ can be derived as

) .407-4) 8cs 4.). The slope of this line on the semi-logarithmic plot below is the maximum specific growth rate.*7-4!. the presence of the substrate induces the synthesis of the "ey enzyme.-M. #ith the substrate no longer present.*7-4- !.(+ 1tarting with a low or zero content of the "ey enzyme needed for growth on a given substrate &representative of poor quality starter culture or inoculum+.! 6. which shows up as a linear increase in the logarithmic plot of cell concentration. emax. The enzyme concentration reaches and stays at its maximum. during the exponential growth phase.&'(. the growth rate or the slope of this curve decelerates and becomes zero when the substrate is completely consumed.) 4./) code0 Calculated !alues of the DE1 !ariables 3ariable initial value minimal value maximal value final value t 5c 5s !! 4 4. mmax . P+.4-7-4) 4.( ) ) !.( 6. 2s the substrate gets depleted.6*7-!! ) !. the "ey enzyme synthesis stops and enzyme degradation during the stationary and death phases reduces the intracellular enzyme content to low levels.! ) 4 4.6*7- e 6. The low or zero enzyme content causes the initial lag phase of no cell growth while increasing enzyme content results in the accelerating growth phase.) 4.) 6.

! !.d&t+ > mu ? 5c <0= d&5s+.4- 4.&:s @ 5s+ .d&t+ > -!.4- 4./967-!4 +DE 2eport 32K*456 .! </= emax > alpha.(664.9 :s !.9 4.47-4) !.4- 4.9 4.(66- 4.4-*7-4) 4./90*)!(.9/6/)9* 4.4-*7- emax !.6/)0((9 (.9 4./967-!4 4.4mumax 4.&mumax@beta+ <(= 7' > e.! 4.emax .! !.47-4) !.d&t+ > alpha ? 5s.47-4) 4.! 4.8cs ? mu ? 5c <*= d&e+.47-4) beta 4.4-*7-4) 7' 4.444! <*= beta > .alpha !.beta?e mu?e 7xplicit equations as entered by the user <!= 8cs > .9 <-= :s > .99(--4) mu 4.4-*7-4) 4) !.ifferential equations as entered by the user <!= d&5c+.) <0= alpha > .4<)= mumax > .

&:s @ 5s+ Figure 2. 5lic" here for the Aolymath 5ode 7. or the growth on the substrate !: 5ells @ 1ubstrate ! &e.).4. which are each capable of supporting cell growth.(.&.ie 5onsider the cell growth medium in a batch bioreactor that contains two different carbon substrates. glucose+ B more 5ells @ products. Dynamic profiles of substrate concentration and intracellular enzyme content along with the logarithm of cell mass concentration.g.<6= mu > mumax ? 7'?5s. 1! and 10. the modified Monod growth rate expression is &'.8 Se9uential Growth : Diau.6+ or growth on the substrate 0: .

5ells @ 1ubstrate 0 &e. consuming the remaining less preferred substrate.! E mmax0. who has termed this phenomenon as the CdiauxieC &Dree" for two growth phases+. the less preferred substrate is not utilized until the preferred substrate is completely consumed and is no longer available in the growth medium. The sequential consumption results in an interesting pattern of the cells first consuming one preferred substrate and then after an intermediate lag phase. mmax.(. lactose+ B more 5ells @ products. This sequential utilization of two substrates in batch cultures has been observed in numerous experiments by Monod. the maximum specific growth rate on mixed substrates: . the modified Monod growth rate expression is &'.). The emzyme growth curve along with the cell and substrate concentration are shown in igure *. i.g. microbial cells do not consume both substrates simultaneously or additively but instead grow on the sugars sequentially. The consistent characteristic of the diauxic growth phenomenon is that the preferred substrate provides the faster growth rate. #hen both the substrates are present in a batch bioreactor.9+ where m1 and m2 are the specific cell growth rates on individual substrates 1! and 10 respectively. . That is. That is.e.

Diauxic growth of bacteria on two substrates. and are determined through maximization of instantaneous growth rate. . The diauxic growth is observed in most or all welldocumented batch cultures on multiple substrates.. rather than an additive growth rate. !96/=. and the evolutionary ob$ective of maximizing the instantaneous growth rate accomplished through hypothesized CcyberneticC variables. The diauxic growth phenomenon has been modeled with the modified Monod equations discussed above. %f the cells were to grow simultaneously on both substrates it has been suggested that the cell growth rate will be reduced to an average of the two growth rates.Figure 3. The preference for faster growth rate in the first growth phase has been suggested to be a consequence of evolutionary pressures on the microbes to grow at the fastest growth rate possible <:ompala et al. The cybernetic variables ui and vi represent the outcome of intracellular regulatory processes controlling enzyme synthesis and activity respectively. showing also the intracellular content of the two hypothetical key enzymes for consuming each substrate.

&'. the diauxic growth phases shown above have been simulated with typical model parameters values.).0 &'.!*+ is the typical dilution of an intracellular species due to cell growth.(.(.(.!0+ for where i > !.). the maximum level of the intracellular "ey enzyme can be shown as &'.!4+ &'. . and The last term in the equation &'.).(. .(.).!-+ #ith these CcyberneticC model equations.!*+ &'.).!!+ &'.).!)+ e.(.uring the exponential growth on high concentration of a single substrate.).(.g.

80 Diau. b-galactosidase for the utilization of lactose+.444! hr! .F for substrate 0. ai > 4. E.F.and e0. these additional model parameters are given some representative values. as long as the instantaneous growth rate m1 is greater than the rate m2.! > 4.4.The model parameters are the Monod growth parameters for growth on each single substrate &which can be determined experimentally+ and additional model parameters ai and bifor the synthesis and degradation of the two "ey enzymes. because of the model parameter values used &mmax.4. 2s the rate-limiting "ey enzymes are hypothetical./) code0 32un this program with S/=** +DE algorithm6 Calculated !alues of the DE1 !ariables . more of the enzymes need for the utilization of substrate ! will be synthesized and active &u1 E u2 and v1 v2 +.* x !4-.ample 2.9 hr-!.hr-!+ and initial conditions for the two "ey enzymes &e!.imi<ation of instantaneous growth rate The cybernetic variables ui and vi represent the optimal response of the enzymatic synthesis and activity to maximize the instantaneous growth of the cells for any given medium composition. 5onsequently cybernetic variables u1 and v1 for the first enzymeHs synthesis and activity remain close to their maximum value of ! during the first growth phase.F for substrate ! and 04 g. mmax. :! > 4.ic Growth0 Ma.8./ hr-!. bi > 4. even though they may be identifiable in many cases &such as.4 at closer to its maximum value of 6..-M. using the initial concentrations of ) g.F.! g. P+.0 > 4. rather than experimentally measured.g. :0 > 4. This is illustrated in the following numerical simulation. Gowever.7. the growth rate on substrate ! m! remains higher than m0 until substrate ! is completely consumed.4 at a lower value of ! x !4-/+. Therefore. even if the substrate ! is present at lower concentration than the substrate 0 at the beginning of batch growth on mixed substrates.

47-4) !.4-*7-4) !.( ) !4 9.4)97-4) 0.4-*7-4) 4) !./ 4.47-4) e!max !./ :! :0 beta! 4.4(07-!! 04 !.47-4) 4.47-4) !.4- !.-!*7-0e! 6.47-4/ !.)4/7-05s0 04 *.- 4.4-*7- !.4.4- 4.*0*7-4- !.*697-4( !.47-4) !.4alpha! !.9 mu0max 4.-*67- ./ 4.*7-4!.4- 4.4- 4.44.! 4 4.9 4.4.47-4) alpha0 !.! 0.4beta0 4.! 4.! 4.3ariable initial value minimal value maximal value final value t 5c 4 4.4.-*67-4) 4) !.-!7-4) 4.( 5s! ) 0.-*67-4) !.4-*7-4) e0max !.! 4.47-4) !.*-*7-4) mu!max 4.47-4) !.4- 4.-*67-4) !.*0*7-4e0 !.9 4./ 4.! 4.9 -0.)4/7-0!4 9.

44!0!4) !6.(4090! 4.d&t+ > -mu0?v0?5c.7! 4.) 4.-*4!00! mumax 4.99/(*)0 4.) ! 4.) 4.90*7-0u! 4.6/*!(0! /.44-)9-( ).4-/)6(9 +DE 2eport 3S/=**6 .44-)/-/ 4.&:!@5s!+?u! beta!?e! .66/*-(6 8cs! 8cs0 v0 ! 4.664/( 4.99)-*)) -!(.96!0*9! mu! 4.!*/7-0mu0 4./90*)!/.) 4.**97-!! 4.) 4./90*)!/.44/4.d&t+ > &mu!?v!@mu0?v0+?5c <0= d&5s!+.!0-6-*70 4.) -!.)/96((9 u0 4.(4090! 4.90*7-0v! ! 4.&mu!?v!@mu0?v0+?e! .6/*!(0! /.8cs0 <)= d&e!+.!0-6-*- 4.) 4.4967-4) ! 4.!*/7-04.44!--06 4.d&t+ > -mu!?v!?5c.ifferential equations as entered by the user <!= d&5c+.8cs! <*= d&5s0+.90*7-04.(664.44*64)9 -0.996(69/ 4.) 4.-)()!)0 /.d&t+ > alpha!?5s!.

mumax ./ <*= :! > .4<(= alpha! > .) <04= 8cs0 > .&mu!?v!@mu0?v0+?e0 7xplicit equations as entered by the user <!= mu!max > .&:!@5s!+ <!)= mu0 > mu0max?70?5s0.<-= beta! > .! <)= :0 > .&:0@5s0+?u0 beta0?e0 .444! <9= e!max > alpha!.<-= d&e0+.444! <6= alpha0 > .&mu!@mu0+ <!(= mumax > if &mu!Emu0+ then &mu!+ else &mu0+ <!6= v! > mu!.d&t+ > alpha0?5s0.4</= beta0 > .&mu!@mu0+ <!/= u0 > mu0.e0max <!*= mu! > mu!max?7!?5s!.&:0@5s0+ <!-= u! > mu!.) <0!= v0 > mu0. mumax <!9= 8cs! > .9 <0= mu0max > .e!max <!0= 70 > e0.& mu!max@ beta!+ <!4= e0max > alpha0.& mu0max@ beta0+ <!!= 7! > e!.

by plotting e!.!.*0*7-4!.e!. The intracellular enzyme levels for the same simulation are shown below to highlight the role of the cybernetic variable u1 and u2 in the synthesis of e1 and e2.4)97-4) .". P+.E e! 6.*7-4!. These are plotted using the last program.-M.B./) /.*0*7-4!. u! and u0 vs time.#$ %imulations of the cybernetic model.5lic" here for the Aolymath 5ode Figure E . showing the profiles for the two substrate concentrations &%# and %2 on the left axis' and logarithm of cell mass &ln ( on the right axis' during a typical diauxic growth.

.)/96((9 u0 4.99)-*)) -!(.996(69/ 4.*-*7-4) 0.e0 !.(4090! 4.".2$ %imulations of the cybernetic model showing the profiles of the two cybernetic )ariables &u# and u2 on the left axis' and two enzymes &e# and e2 on the right axis' . triggering the switch in the cybernetic variables and inducing synthesis of enzyme 0.uring the first growth phase.44!0!4) !6. the growth rate m1 on that substrate falls to zero.44-)/-/ 4.*697-4( !.47-4/ !.!.-!7-4) u! 4. 2fter the first substrate is mostly consumed. indicating preferential synthesis of enzyme ! and repression &suppression of synthesis+ of enzyme 0 as u2 is close to zero.(4090! 4. the cybernetic variable u1 ta"es values close to unity.-*4!00! Figure E .

ample 2.! 4 4. along with significant consumption of 10 during the first growth phase.)!(7-!) !.ic Growth0 Effect of Preculturing in Substrate > %t may be suspected from the above simulation that preferential utilization of substrate ! is due to the high level of enzyme ! assumed as its initial value. The diauxic lag gets significantly shortened. the initial value for enzyme 0 should be higher and that for enzyme ! should be assumed much lower. 0 Diau.>.( 5s! ) !.4.7./) code0 Calculated !alues of the DE1 !ariables 3ariable initial value minimal value maximal value final value t 5c 4 4.! !.-M.)!(7-!) !4 . This high initial value for e1 is chosen to indicate preculturing the inoculum in substrate !.E.((-7-!!4 -. substrate ! is gradually preferred with increasing culture time and is completely consumed during the first growth phase.8. Ievertheless.( ) !4 -.((-7-!5s0 !4 !. while "eeping all other initial values and model parameter values identical to those in the above example. %f the inoculum is precultured in substrate 0. The two figures below show simulation results with the altered initial conditions for the two enzymes. P+.

! 4.9!/-(9( mu! 4./*/7-!) u! 4.4-*7-4) e0max !./*-(09 4.!/7-4) mu!max 4.449- 4.47-4) !.! 4.4- 4.4494.4-*7-4) 4) !.0)67-!mu0 4.(-)6- !.0)67-!4.47-4) !./ :! :0 beta! 4.446*)!/.-*67-4) !.)(60*/6 /./ 4.47-4) alpha0 !.4.40!9/)4.*(!)06/ !.06)7-4- !.-00-(49 !.49/7-4e0 !.9 !.4- !.)!7-4) 4.-*67- 4./4.47-4/ /./ 4.0(/*(9* .! 4.47-4) !.- 4.! 4.4- 4.-*67-4) 4) !.-*67-4) 7! 4.47-4) !.4-*7-4) !.47-4) 4.4-*7- !.4.9 mu0max 4./49!(9/ 4.47-4) e!max !.)(*)-*! 4.9 4.4beta0 4./ 4./*/7-!) 4.)!07-4- (.4- 4.e! !.4alpha! !.09)*-9) 70 4.4- 4.9 4.47-4) !.47-4) !.4!*)/!* 4.47-4/ *.4.44.

u0 4.9(64*-4.*/)0(! 4.96/-*6( 4.(0*/04( mumax 4.-00-(49 4.*(!)06/ !./*/7-!) !./*/7-!)

v! 4.400)-(6 4.*6!9*9) 8cs! 8cs0 v0 ! 4.) 4.) !

4.4!*/)-

!

4.) 4.) 4.-(099()

4.) 4.) !

4.) 4.)

+DE 2eport 3S/=**6

;ifferential equations as entered by the user <!= d&5c+,d&t+ > &mu!?v!@mu0?v0+?5c <0= d&5s!+,d&t+ > -mu!?v!?5c,8cs! <*= d&5s0+,d&t+ > -mu0?v0?5c,8cs0 <)= d&e!+,d&t+ > alpha!?5s!,&:!@5s!+?u! beta!?e! - &mu!?v!@mu0?v0+?e! <-= d&e0+,d&t+ > alpha0?5s0,&:0@5s0+?u0 beta0?e0 - &mu!?v!@mu0?v0+?e0

7xplicit equations as entered by the user <!= mu!max > .9 <0= mu0max > ./ <*= :! > .! <)= :0 > .<-= beta! > .4-

</= beta0 > .4<(= alpha! > .444! <6= alpha0 > .444! <9= e!max > alpha!,& mu!max@ beta!+ <!4= e0max > alpha0,& mu0max@ beta0+ <!!= 7! > e!,e!max <!0= 70 > e0,e0max <!*= mu! > mu!max?7!?5s!,&:!@5s!+ <!)= mu0 > mu0max?70?5s0,&:0@5s0+ <!-= u! > mu!,&mu!@mu0+ <!/= u0 > mu0,&mu!@mu0+ <!(= mumax > if &mu!Emu0+ then &mu!+ else &mu0+ <!6= v! > mu!, mumax <!9= 8cs! > .) <04= 8cs0 > .) <0!= v0 > mu0,mumax

Figure E .!.".3$ %imulations of cybernetic model with altered initial conditions for the two enzyme le)els, with e2 * e#, reflecting the preculturing of inoculum on %2.

The gradual increase in the slope of semi log plot of cell concentration during the first growth phase is due to the gradual preference of substrate !, even though the inoculum is precultured on substrate 0 and has the enzyme 0 already available for its continued consumption. ;uring the later parts of first growth phase, more of the enzyme ! is synthesized &as is seen in the next simulation graph+ resulting in the rapid consumption of substrate ! and increasing growth rate &slope of the semi log curve+. 1ignificant availability of enzyme 0 at the end of first growth phase results in the reduced or non-existent diauxic lag phase before the second growth phase on the remaining substrate 0. P+,-M./) /.B,E0 e! !.47-4/ *.49/7-4e0 !.47-4) !.!/7-4) !.47-4/ /.)!07-4-

(.06)7-4-

!.)!7-4)

!.*/)0(! 4.".u! 4.4!*)/!* 4.(0*/04( 4.9(64*-4.4. 2." %imulations of the cybernetic model showing the two cybernetic )ariables u1 and u2 along with the profiles of intracellular enzyme contents for the two key enzymes e1 and e2. E)en with the )alues.96/-*6( Figure E . the batch culture .0(/*(9* u0 4./*-(09 4. the model predicts an increasing preference for the substrate # during the first growth phase. +reculturing causes the initial )alue for e2 to be much higher than that for e1.40!9/)4.7.> Simultaneous tili<ation in Continuous Cultures or continuous or chemostat culture of microbial growth on multiple substrates.

(. %n equation '.(.!9+ where .).!/+ &'. is the dilution rate.).0 &'. an additional synthesis rate constant a? is included to ensure a low level presence of each enzyme even in the absence of its substrate. 2t even higher &than the maximum growth rate possible in .(. as observed experimentally. the cybernetic model predicts the simultaneous utilization of both substrates at steady state for low dilution rates.).balance equations given above can be modified to include the inlet and outlet terms as below: &'. #ith these new dynamic balance equations for continuous cultures.!(+ &'.).(.(.!6+ !o" i > !. 2t increasing dilution rates.).!9 for intracellular enzymes.4 are the inlet concentrations of substrates 1! and 10 respectively. 51!. the simultaneous utilization of both substrates changes gradually to #"e!e"ential utilization of the preferred &faster growth supporting+ substrate.4 and 510.

The inlet concentrations of the two substrates are chosen as !4 g.(.4!a+.>..-M.ample 2.8 Simultaneous utili<ation of multiple substrates in continuous cultures . ai > 4. The initial values for the different concentrations are immaterial &as long as cell mass concentration is not started at zero since there will be no spontaneous generation of life in a sterile bioreactor+ if we simulate the dynamic balance equations &'.(.sing the same model parameter values used in the previous examples &mmax.hr-! and the new parameter ai?> 4./ hr-!. mmax.e.! > 4.).444! hr-!.4. :! > 4.!/-'.d&t+ > &mu!?v!@mu0?v0+?5c .!9+ long enough for them to reach steady state. E.F. washout of cells from the chemostat is observed as also observed for single substrate chemostats./)6 +DE 2eport 3S/=**6 .ifferential equations as entered by the user <!= d&5c+.7. cell mass concentration in the feed is zero+.8cs! @ . P+.-M. the above modified cybernetic model equations of continuous cultures can be simulated to plot the steady state concentration of cell mass. :0 > 4.! g.F.g.0 > 4./) code0 3?ote0 Sol!e this program using the S/=** algorithm in P+.4.?5c <0= d&5s!+.the chemostat: + dilution rates.?&5s!45s!+ .9 hr-!.F each and the inlet nutrient feed is assumed to be sterile &i.. substrates ! and 0 over a range of dilution rates.d&t+ > -mu!?v!?5c. bi > 4.).

9 <0= mu0max > .&:!@5s!+?u! beta!?e! ./ <*= :! > .&mu!@mu0+ <!(= mumax > if &mu!Emu0+ then &mu!+ else &mu0+ <!6= v! > mu!.8cs0 @ .&:0@5s0+?u0 beta0?e0 .<-= beta! > .&mu!?v!@mu0?v0+?e! @ alphastar <-= d&e0+.&mu!?v!@mu0?v0+?e0 @ alphastar 7xplicit equations as entered by the user <!= mu!max > .&:0@5s0+ <!-= u! > mu!.d&t+ > alpha0?5s0. mumax .e!max <!0= 70 > e0.444! <9= e!max > alpha!.444! <6= alpha0 > .& mu!max@ beta!+ <!4= e0max > alpha0.&:!@5s!+ <!)= mu0 > mu0max?70?5s0.e0max <!*= mu! > mu!max?7!?5s!.&mu!@mu0+ <!/= u0 > mu0.? &5s04 5s0+ <)= d&e!+.<*= d&5s0+.! <)= :0> .d&t+ > alpha!?5s!.4</= beta0 > .d&t+ > -mu0?v0?5c.& mu0max@ beta0+ <!!= 7! > e!.4<(= alpha! > .

49469)* 5s0 04 !9.6/-7-4- !.mumax <00= alphastar > .47-4/ !.! !.<!9= 8cs! > .9/!)0! e! 6./.-6494-* 4.! to . .4!?alpha! <0*= 5s!4 > ) <0)= 5s04 > 04 <0-= .4-*7-4) !. > .). > .) <0!= v0 > mu0.*7-49.*7-4- !. values. > .! and collect the final values &equilibrium values+ and plot them against the corresponding .and .! !4 !4 5c 4.9 with intervals of .) <04= 8cs0 > .6/-7-4- .4 3ariable initial value minimal value maximal value final value t 4 4 4. Diven are the tables for .. Calculated !alues of the DE1 !ariables for D @ .-6494-* 5s! ) 4./9)7-4e0 !.46!/(-! ) !9.) #e run this program with .47-4/ !. > .9/!)0! 04 6.

4- 4.44/- 4.4- !.47-4) 4.44*64)9 4.47-4) !.9 4.99)-*)) 4.44/4.47-4) alpha0 !.! 4.47-4) !.*9)(/-! 4.!!990/9 !.!-49(mumax 4.44*64)9 4.6(006/( v! ! 4.9 mu0max 4.44-)/-/ 4.!-49(4.-*67-4) 4) !.4-*7-4) e0max !.4alpha! !.47-4) !.! 4./ 4./ 4.4- 4.! 4.mu!max 4.47-4) !.99)-*)) 4.4beta0 4.*9)(/-- ! ! .-*67- 4.- 4.4.(66- !.4(4!9(9 u! 4.6(006/( 4.9 4./ :! :0 beta! 4./90*)!4.47-4) e!max !.*9)(/-mu0 4.4-*7-4) 4) !.6)940u0 4.-*67-4) !.4-*7-4) !.4- 4.4- 4.44444(0 4.90!(-** 70 4.-*67-4) 7! 4.6)9404.!!990/9 mu! 4.4- 4.*9)(/-4.(664.4-*7- !.47-4) !.44-)/-/ 4.4.9 4./ 4./90*)!4.! 4.- 4.4(4!9(9 4.

-)699(9 5s! ) 4.) v0 4.) 4./ 4.!094-4/ ) !9.! 4.4.) 4.! !.!((60!/ alphastar !./ 4.47-4/ 5s!4 5s04 .47-4/ -.44-)9-( 4.) ) 04 4.99(*! 04 6.) 4./407-4/ mu!max 4.47-4/ !. !.!*!6(99 5s0 04 !9.4-*7-4) !.) 4.) 4.99(*! e! 6.*7-4- !.) 4.) ) 04 4.9 !.- 4.9 mu0max 4.- 4.! 4.! 4.5 3ariable initial value minimal value maximal value final value t 4 4 4.) 4.*7-4!.! 4.9 4./ 4./407-4/ 4./ :! :0 4.9 4.47-4/ ) 04 4.47-4/ !.!((60!/ 4.! !4 !4 5c 4.- .-)99-(( 4.) Calculated !alues of the DE1 !ariables for D @ .40/7-4) e0 !.47-4/ -.) ) 04 4.8cs! 8cs0 4.44-)9-( 4.

9-9*)(4.4- 4.4)0*(-! 4.47-4/ .4-*7- !.(664.) v0 4.47-4) alpha0 !.47-4) !.47-4) !.99)-*)) 4.(66- !.4)4/-04.) 4.47-4) e!max !.4)4/-0mumax 4.)96(99( ! 4.44-)/-/ 4.47-4) !.44/4.) ! 4.44-)9-( 4.4- 4.4-*7-4) 4) !.47-4/ !.44-)/-/ 4.4*/!46/ mu! 4.9()//0/ 70 4./90*)!4.6(*/!0) 4.4- alpha! !.4beta0 4.) 4.47-4/ !.beta! 4.4)0*(-! alphastar !.47-4/ !.99)-*)) 4.9-9*)(u0 4.4-*7-4) e0max !.44/- 4.4-*7-4) !.4- 4.4- 4.-*67-4) 7! 4.47-4) !.40!!*/( 4.-*67-4) 4) !.44*64)9 4.444!0)( 4.47-4) !.40!!*/( u! 4.4*/!46/ !.-*67-4) !.)96(99( mu0 4.) 4.) 4.4- 4.-*67- 4.4- 4./90*)!4.44*64)9 4.)96(6/ ! 4.) 4.6(*/!0) v! 8cs! 8cs0 ! 4.) 4.47-4) !.)96(6/ 4.44-)9-( 4.

! 4.! 4.4.999(!/ 04 6.*7-4- !.- ) 04 4.!/*7-4/ 4.44.4-*7-4) !.!/*7-4/ mu!max 4.! !.A Calculated !alues of the DE1 !ariables 3ariable initial value minimal value maximal value final value t 4 4 4.9 4.! !4 !4 5c 4.- 4.0(/-)!* 4.4- 4.649--46 5s0 04 !9.4.47-4/ 0.9 4./ 4.649--46 ) !9.4beta0 4.! 4.9 !.- ) 04 4.4.*7-4!.4- ./ 4./ :! :0 beta! 4.0(/-)!* 5s! ) 4.999(!/ e! 6.9 mu0max 4.- Calculated !alues of the DE1 !ariables for D @ .4- 4.5s!4 5s04 . ) 04 4.- ) 04 4.! 4.47-4/ 0.44.4- 4.4-!7-4) e0 !./ 4.4- 4.

4!)4)!( !.99)-*)) 4.47-4) !.47-4) !.47-4/ !.4-*7-4) !.-*67-4) 4) !./ ) 04 4.99)-*)) 4.4!4!-9( mumax 4.47-4/ !.44-)9-( 4./ ) 04 4./ ) 04 4.4-*7- !.) 4.47-4/ 5s!4 5s04 .44/- 4.44/4.-*67-4) 7! 4.4460!9u! 4.47-4/ ) 04 4.44-)/-/ 4.) 4.44*64)9 4.-*67-4) !./90*)!- ! 4./90*)!4.) 4.4!40/) alphastar !.6()(/9* v! 8cs! 8cs0 ! 4.) 4.4460!94.) 4.9696)4* 4.47-4) !./90*)!4.44404)! 4./ .44*64)9 4.996!9/! 70 4.47-4) e!max !.4!)4)!( mu! 4.(664.4!4!-9( 4./90*)!4.47-4) !.9696)4* u0 4.4-*7-4) 4) !.) ! 4.(66- !.44-)/-/ 4.47-4) alpha0 !.6446!-6 ! 4.) 4.) v0 4.4-*7-4) e0max !.4!40/) 4.-*67- 4.44-)9-( 4.alpha! !. !.47-4) !.6446!-6 mu0 4.6()(/9* 4.47-4) !.

%teady state )alues from the simulation of cybernetic model e-uations. .5lic" here for the Aolymath 5ode Figure E . /t higher dilution rates.oth substrates are simultaneously utilized at low dilution rates. washout occurs with the steady state )alues same as the inlet concentrations.". substrate 2 is gradually re0ected in fa)or of substrate #.!. .. /t increasing dilution rates.

maximization of the cell growth rate through preferential utilization of a substrate or in this case a metabolic pathway over the others. if glucose consumption for cell growth is ignored: & . which may be represented by the overall chemical equation. the continuous culture simulations &and the experimental data+ show the simultaneous utilization of both the substrates at low dilution rates. Saccha"omyces ce"evisiae.".& Multiple Metabolic Pathwa"s in -east0 The brewerHs or ba"erHs yeast.7.e. the washout steady state is observed with the two substrates and the cell mass reaching a steady state that is the same as their inlet concentration. 2t increasing dilution rates.21' The Monod growth parameters for this growth process are: .%n mar"ed contrast to the batch culture results of sequential utilization of the two substrates. 2.!.4. presents an interesting example of the cybernetic ob$ective i.e. faster growth rate supporting+ substrate. 2t much higher growth rates. the second &less preferred or lower growth rate supporting+ substrate is not consumed completely or is re$ected in favor of the preferred &i. The yeast cells have different pathways for consuming glucose: $1% &lucose !e"mentative #ath'ay.

(2) Ethanol oxidative pathway. The Monod growth parameters for this third growth process are: and $(% &lucose oxidative #ath'ay.2#' can also occur if ethanol and oxygen are both present in the culture medium. again ignoring the glucose consumption for cell growth.".!.!. which is of course possible only in the presence of oxygen. with its o)erall chemical reaction &ignoring cell growth'$ & .". the overall chemical reaction of this pathway can be represented as: & .22' The Monod growth parameters for this second growth process are: .

0*+ . and if latter.(. which pathway is preferredJ Iumerous brewers routinely ferment glucose to ethanol using yeast cells. > 4+ and continuous cultures. These fermentations are successful &in producing ethanol+ because cells preferentially use the !aste" fermentative pathway and do not produce the enzymes needed for slo'e" oxidative consumption of glucose even if oxygen is present in culture medium. which will occur in a subsequent or diauxic growth phase if oxygen is present.). 2fter all glucose is fermented. The cybernetic model equations introduced earlier predicts the diauxic growth of yeast on glucose and ethanol in aerobic cultures. represented as 5T+ from Kones and :ompala &!999+ are given below for both batch &.)y#othetical *uestion: %n a typical brewing experiment. are both pathways used simultaneously or is one pathway preferentially utilized by the yeast. if glucose and oxygen are both present in the culture medium. &'. without ta"ing any special precautions to eliminate oxygen from the culture medium. trehalose and glycogen. The further modified cybernetic model equations for the yeast growth metabolism &to include the dynamics of intracellular storage carbohydrates. with small modifications to incorporate the specific case of ethanol generation from the fermentative pathway. it will be necessary to stop the batch fermentation to avoid the oxidative consumption of ethanol. until almost all the glucose has been fermented to ethanol.

(.).(.0)+ &'.(.(.).(.).0-+ &'.09+ &'.*4+ &'.).).&'.).(.).(.06+ 2he modified 3onod growth rates along the indi)idual metabolic pathways are$ &'.*!+ .0/+ &'.0(+ &'.(.).

4./) code Calculated !alues of the DE1 !ariables 3ariable initial value minimal value maximal value final value t 4 4 4.47-4/ 4. P+. The model parameters were chosen to fit the experimental data from von Meyenburg &!9/9+ and are partially listed earlier with discussions on the three metabolic pathways.*(6) 5g ) -!..*0+ The symbols ! and +represent the stoichiometric constants for the production of ethanol.*/*!)6( !.*7-44.47-4/ 4.4/*/)* e! 6.*7-4- 4.4/*/)* 4. The cybernetic model for yeast metabolism predicts the diauxic growth phases in the aerobic growth of yeast on glucose in batch cultures &with .*(6) -!.0()* 5o 4./-7@4) ) 04 (94-.*4/)0)) e0 !.!4-4!!9 0//9.!4(949* .5ybernetic variables ui and vi are determined as before: &'.a+.0()* 4.(. consumption of oxygen and the storage carbohydrates respectively.! 04 04 5c 4.-M.! 0//9.- 6. > 4 and high values of -./-7@4) 5e 04 (94-.).

( 4.)) 4.* e*max 4.0/*!-(9 4.0/*!-(9 4.46!))!) 5t 4 -.( 4.!9 4.**(4(6( 4.*64)/() !.-**7-4) *.6 mu!max 4.!0!449) 4.0/*!-(9 e0max 4.* 4.* 4.4! 4.0/*!-(9 4.e* !.( 4.06((-9/ 7! *.!-)7-4) !.47-4) 4.* 0.0 4.* 4.( 4.* 0.*/ alpha* :o* alpha! alpha0 4.( 4.!9 4.**(4(6( 4.( 4.06*4!69 e!max 4.* 4.6 4 !.06*4!69 4.6 4.4! 4.( 4.47-4) 4.( 4.06*4!69 4.06*4!69 4.4! beta0 beta* mu0max 4.* 0.)) 4.*/ 4.( 4.!9 mu*max 4.* 4.*/ 4.))4-)64.0 4.!/))!06 .!-)7-4) *.!9 4.**(4(6( 7* *.)) :o0 4.( 4.0 4.* 4.(697-4( beta! gam* 4.( 4.( 4.0 4.* 0.)) 4.**(4(6( 4.6 4.4! 4.* 4.-**7-4) 4.!0)()*) 4.*/ 4.

440)600 alphastar 4.(- 4.)/!!4!! -4.*(!7-4) 4.(- 4. 4 4.*6**(9( 4./ 4.70 0.!/ 80 4.9/(7-4/ 0.!/ 4.44! mu! !.9!!)(() u0 4.4--0/6! 4.)/!!4!! -9.*!!-*-* :! 4.-*/(*/ 4.!/ 4.! 4.44! 4.4! :* 4.! !4 04 4 !4 04 4 !4 04 4 !4 04 .4! 4.4! 4.! 5g4 5s04 .-0/4*09 9./ 4.4- 4.*(!7-4) !./ 4.440!/09 4.!/ 4.!((7-4) -!.49!44)6 8! 4.(- 4.(8* 4.9/(7-4/ 4.44! 4.64!()-( 4.! 4.44! 4.44.4! 4.!00/696 4.440!/09 4.-0*7-4( 4.4:0 4.-!0*)*0 !.*(!7-4) u! 4.!964-66 !.44!*9-0 mumax !.-!0*)*0 mu0 -.-0*7-4( mu* !./ u* 4.44.*04!*46 4.*(!7-4) !.*4))/0* -4.!6-!0-) 4.!964-66 4.4-!!-) -.

*607-40.)6 phi0 phi* phi) 4.ifferential equations as entered by the user <!= d&5c+.4907-4-4.d&t+ > .*607-4) d5c 0.! 4.66)9!4) 4.06!gam! !4 gam0 !4 !4 !4 !4 !4 !4 !4 d5t 6.!*-9)*! 4.! 4.phi)?&5t?d5c @ 5c?d5t+ .8!@ mu*?v*.v* 4.-!()-)) 0.9- 0 ! 4.9- 0 ! 4.-0.)6 4.44)4096 4.06!!*6!.! 4.6-946*( -4.d&t+ > d5c <0= d&5g+.)6 4.*607-4!*6!.9- "la !444 5ostar !444 !444 !444 4.44)4096 4.440(0** v! ! ! ! ! v0 4.9*(4946 4.---7-4( phi! 4.0-!9-) 0.*607-4) 4.! +DE 2eport 32K*456 .8*+?5c .6-946*( -4.5g+ .)6 0 ! 4.4996)*0 sigmamuv 0.&mu!?v!.?&5g4 .!!)*/(.9- 0 ! 4.

( <(= mu0max > .d&t+ > d5t 7xplicit equations as entered by the user <!= beta! > .d&t+ > "la ? &5ostar .( </= beta* > .d&t+ > -.?5e @ & phi!?mu!?v!.4! <-= beta0 > .* <!4= :o* > 0.d&t+ > alpha!?5g.8! mu0?v0.5o+ &phi0?mu0?v0.*/ <9= alpha* > .( <0= gam* > .& mu!max@ beta!+ .6 <*= mu!max > .0 <!!= alpha! > .&:*@5g+?u* & beta*@ sigmamuv+ ?e* @ alphastar <6= d&5t+.80+ ? 5c <)= d&5o+.&:!@5g+?u! & beta!@ sigmamuv+ ? e! @ alphastar </= d&e0+.!9 <6= mu*max > .* <!0= alpha0 > .d&t+ > alpha0?5e.8*+ ? 5c <-= d&e!+.* <!*= e*max > alpha*.& mu*max@ beta*+ <!)= e!max > alpha!.<*= d&5e+.&:0@5e+?u0 & beta0@ sigmamuv+ ?e0 @ alphastar <(= d&e*+.d&t+ > alpha*?5g.80 @ phi*?mu*?v*.)) <)= :o0 > .

&mu!@mu0@mu*+ <0(= u0 > mu0.e*max <!(= 7! > e!. > 4 <*/= v* > mu*. mumax .& mu0max@ beta0+ <!/= 7* > e*.(<*4= 8* > .&:o* @ 5o+ <0-= mumax > if &mu!Emu0+ then &if &mu!Emu*+ then &mu!+ else &mu*++ else &if &mu0Emu*+ then &mu0+ else&mu*++ <0/= u! > mu!.mumax <*(= v! > mu!.! <**= 5g4 > !4 <*)= 5s04 > 04 <*-= .&mu!@mu0@mu*+ <*0= alphastar > .&:0@5e+ ? 5o.e0max <!9= :! > .<!-= e0max > alpha0.44! <00= mu! > mu!max?7!?5g.4<04= :0 > ./4 <*!= u* > mu*.!/ <09= 80 > .e!max <!6= 70 > e0.&:o0 @ 5o+ <0)= mu* > mu*max?7*?5e.4! <0!= :* > .&:*@5g+ ? 5o.&:!@5g+ <0*= mu0 > mu0max?70?5e.&mu!@mu0@mu*+ <06= 8! > .

9<)6= "la > !444 <)9= 5ostar > .mumax <*9= sigmamuv > mu!?v!@mu0?v0@mu*?v* <)4= d5c > &sigmamuv .<*6= v0 > mu0.! 5lic" here for the Aolymath 5ode ..+?5c <)!= gam! > !4 <)0= gam0 > !4 <)*= d5t > gam*?mu*?v* &gam!?mu!?v!@gam0?mu0?v0+?5t sigmamuv?5t <))= phi! > .)6 <)-= phi0 > 0 <)/= phi* > ! <)(= phi) > .

0. to get various concentration plots of the cell and the substrates.) P+. as seen from the high cell mass yields in igure . followed by switch to the fermentative pathway at higher dilution rates. 2t higher dilution rates. and set different values for .*. is the preferred pathway for glucose consumption at the low dilution rates. the yeast cells clearly prefer the faster fermentative metabolism and ignore or repress the oxidative metabolism.igure ). #e can use the earlier program. glucose. . . .E0 Calculated !alues of the DE1 !ariables 3D @ .uring the first growth phase. Diven are the tables of values for . 5ybernetic model simulations and experimental data from von Meyenburg &!9/9+ for cell mass. the utilization of glucose fermentative pathway is seen both in low cell mass yield in igure . > . 2fter glucose is completely fermented.-M./) /. >6 . The oxidative consumption of glucose. which is not utilized in the batch aerobic cultures.and absence of any ethanol production.B.as well as the production of ethanol &data not shown+. This choice of the fermentative pathway can be concluded from &!+ the growth rate &the slope of a semi-long plot of cell mass+ during the first growth phase or more easily &0+ the accumulation of the fermentation product. ethanol. the presence of oxygen enables further growth of yeast cells in a second or diauxic growth phase using ethanol oxidative pathway. The yeast cybernetic model equations for continuous or chemostat cultures can be simulated to predict the C5rabtree effectC of preferential utilization of glucose oxidative pathway during the low dilution rates. ethanol concentrations in aerobic batch culture of Saccha"omycesce"evisiae.

( 4.*0!( 5g ) -004.( 4.47-4/ 4.47-4) 4.4! 4.( 4.47-4/ 4.*-0*040 !!4.4! 4.4! 4.)) 4.46!!)!) 5t 4 !.( 4.6 -004.!9 4.( 4.( 4.4! beta0 beta* mu0max 4.( 4.*67-4/ beta! gam* 4.*7-4- 4.*/ 4.)6(6( 5o 4.!9 mu*max 4.)) 4.6 4.499)06* 4.47-4) 4.)6(6( 4.- 6.4.49/*)/9 04 *(.!466*( !.(4*)) /.( 4.!004*4* /.)) :o0 4.!40!9*- 4 4.*7-44.!9 4.( 4.!4-60*9 e* !.6 mu!max 4.3ariable initial value minimal value maximal value final value t 4 4 04 4.499)06* e! 6.4*/4-90 4.)) 4.*/ .(4*)) 5e 04 !!4.*/ 4.( 4.( 4.*4-90( e0 !.! *(.*4(6-)( !.*0!( 5c 4.6 4.*/ 4.( 4.!9 4.

*(!7-4) 4.44! 4.!-)7-4) *.!/!/)0/ 4.4- 4.4! 4.*/!)9(( 4.!!-9!-0 -4.0/*!-(9 4.* e*max 4.4! 4.44.4--/6*( 4.* 4.alpha* :o* alpha! alpha0 4.-!!/0-9 !.94(64-6 4.* 4.4400*)0 mumax 4.-**7-4) *.* 0.0/*!-(9 4.06//996 7! *.4:0 4.0/*!-(9 4.0*!(*/! .*006(-) 4.-*/(*/ !.4! :* 4.-0*7-4( 4.* 0.-**7-4) 4.**(4(6( 4.*!*9))* :! 4.!/0-00( 70 0.!((7-4) -4.4-)!9*mu* !.**(4(6( 7* *.**(4(6( 4.*(!7-4) !.* 4.*(!7-4) 4.0 4.4! 4.!/96)6! !.0/*!-(9 e0max 4.-*/(*/ 4.-(60)/) !.-0*7-4( -.0 4.* 4.* 4.9/(7-4/ 4.44.06*4!69 4.0 4.* 0.06*4!69 4.9/(7-4/ 0.44! 4.44! 4.-(60)/) 4.-!!/0-9 mu0 -.* 4.0 4.* 0.06*4!69 4.*(!7-4) u! 4.44! mu! !.**(4(6( 4.!-)7-4) !.06*4!69 e!max 4.* 4.

-!(*(/ 0.! 4.-).)6 0 0 0 0 . 4.6-946*( -4.49/!-68! 4.4907-4-4.440!/09 4.4!99(/0 -4.*607-4) 4.!/ 4.*))*!6( 4.44)*//9 v! ! ! ! ! v0 4.*607-4) d5c -4.0 !4 04 4.(- 4.! !4 04 4.!4-90)! sigmamuv 4.! 5g4 5s04 .0 !4 04 4.u0 4.6)-4!* gam! !4 gam0 !4 !4 !4 !4 !4 !4 !4 d5t 6.)/!!4!! -4.! 4.6-946*( -4.44)4096 4.!!0-60/ 4./ 4.!/ 4.(- 4.)6 phi0 4./ u* 4.0(9-*9* 4.440!/09 4.9)/7-46 phi! 4.(8* 4.)6 4.4!99(/0 !!./060/0( 0.6)-4!* !!.!*064( 4.)/!!4!! -4.0 v* 4.0 !4 04 4.!/ 4.4!(940.44*9/)* alphastar 4.!9(6*// 4.)6 4.(- 4.!/ 80 4./ 4./ 4.44)4096 4.

/*/(9)! 04 4.47-4) 4.6 4.94(7-4/ beta! gam* 4.49!)9!6 04 ).!0)64! 4 4.! 4. &6 3ariable initial value minimal value maximal value final value t 4 4 04 4.4-!9(-- 4.! Calculated !alues of the DE1 !ariables 3D @ .496*6/ 4.6 4.47-4/ 4./9*6*/ (.6)60)4/ 5c 4.47-4/ 4./9*6*/ 5e 04 !).9- ! !444 !444 !444 4.!(/)(! 5o 4.( 4.! 4.6 -!9.464)!(5t 4 0./!/464! 0.6)60)4/ 5g ) -!9.( 4.- 6.4.*4//*99 e0 !.9- ! 4.( .9- ! 4.6 4.*7-4- 4.! ).9"la !444 5ostar ! 4.! 4.49990/( e! 6.!!4*!)* !.phi* phi) 4.*4(0-/* !.*7-44.( 4.47-4) 4.!4-*)6e* !.

* 4.**(4(6( 7* *.4! 4.)) 4.* 4.0 4.!/-0*!/ 70 0.-**7-4) 4.*/ 4.( 4.)) 4.( 4.9/(7-4/ 0.!9 4.* 4.4! 4.0 4.0 4.!/(-(** !.4! beta0 beta* mu0max 4.06*4!69 e!max 4.))4)49 7! *.* 0.* e*max 4.4:0 4.0 4.( 4.*!0-**9 :! 4.* 4.* 4.!-)7-4) !.-**7-4) 4.4! 4.0/*!-(9 e0max 4.4! 4.4! 4.)) :o0 4.06)!)!( 4.06*4!69 4.4! .4- 4.0/*!-(9 4.* 4.( 4.*/ 4.*/ 4.**(4(6( 4.( 4.06*4!69 4.)) 4.**(4(6( *.0/*!-(9 4.!9 mu*max 4.* 0.!-)7-4) *.*0(0/4( 4.mu!max 4.**(4(6( 4.44.44.* 0.4! 4.( 4.*/ alpha* :o* alpha! alpha0 4.06*4!69 4.* 0.9/(7-4/ 4.!9 4.( 4.0/*!-(9 4.!9 4.* 4.( 4.

)/!!4!! -4. 4.! 5g4 5s04 ./(0*!4) -4.* ! !4 04 4.44)4096 4.(- 4.(8* 4.4*96!00 4.* !4 04 4.9!4!-!! u0 4.-!)44/9 !.44/00)) v! ! ! -4.-0*7-4( -./ 4.*(!7-4) u! 4./ u* 4.!/ 80 4.!46(9)* 4.*(!7-4) 4.!/ 4.44-//-! alphastar 4.44! mu! !.*94(!/0 ! v0 4.*(!7-4) !.!/ 4.-//09(( -4.440!/09 4.(- 4.! !4 04 4.49--!)! 8! 4.44! 4.!4)9)*! .44*!99) mumax 4.4-/*9(9 4.44)4096 4.44! 4.0/0/60!.:* 4.-!)44/9 mu0 -.!/ 4.* !4 04 4.-*/(*/ 4.! 4.* v* 4.-!(/-4/ 4.440!/09 4.44! 4.!((7-4) -4./ 4.!-/(09( 4.4-*9)!mu* !.!)6/4/4.4(!60-6 4.! 4.(- 4.6-946*( -4.*(!7-4) 4./ 4.-//09(( !.-0*7-4( 4.!)09!/( 4.

sigmamuv 4.)6 0 ! 4.! 4.9- 0 ! 4./-64)0( 0.! 4./)0*4/* 5c 4.! Calculated !alues of the DE1 !ariables 3D @ .4099(/0 -4.94!9*0- ) 6.*607-4) 4.4/-496* !.9*06(-6 04 .6*)7-4( phi! 4.! 4.4099(/0 !.9- 0 ! !444 !444 !444 4.*607-4) d5c -4.4/-496* gam! !4 gam0 !4 !4 !4 !4 !4 !4 !4 d5t 6.9- 0 ! 4.9"la !444 5ostar 4.46*9)*) 04 4.009*-*6 4.4907-4-4./)0*4/* 5g ) /.)6 4.)6 4.)6 phi0 phi* phi) 4.-!9/6(/ 0.409*-09 5e 04 !.! 4. 56 3ariable initial value minimal value maximal value final value t 4 4 04 4.*(!6/-6 -/.()9/()* 4.

( 4.!4/(49! !.06*4!69 4.*4)!)( e0 !.* e*max 4.*/ 4.!9 4.06*4!69 4.* 4.46)9*- 4 /.( 4.( 4.( 4.( 4.06*4!69 e!max 4.0/*!-(9 4.* 4.4.47-4/ 4.0/*!-(9 4.*7-44.-6(7-4( beta! gam* 4.* 0.47-4) 4.)) 4.* 0.47-4) 4.46*!/6/ 5t 4 -.49999 4.* 4.* 0.*/ alpha* :o* alpha! alpha0 4.!4/))-! e* !.* 0.* 4.0 4.!9 4.( 4.0 4.( 4.- 6.!9 mu*max 4.*4)(!6! !.( 4.06*4!69 4.0 4.4! beta0 beta* mu0max 4.*/ 4.4! 4.0/*!-(9 4.0/*!-(9 .)) 4.* 4.!9 4.*7-4- 4.5o 4.6 mu!max 4.4! 4.6 4.*/ 4.6 4.* 4.49999 e! 6.)) 4.( 4.( 4.)) :o0 4.( 4.4! 4.6 4.( 4.0 4.47-4/ 4.* 4.!/!7-4- 4.

4!*7-4) 4.44! 4.44.*!-(6(0 :! 4.!-)7-4) *.4- 4.440-9 .44666/( 4.!--(-6/ 70 0.4! :* 4.49(9(0( 4.94*0//* 4./ 4.44! mu! !.!/ 80 4.49/66! 8! 4.!((7-4) !.44! 4.*(!7-4) u! 4.!/ 4.-4)*-!* mu0 -.*(!7-4) 4.440!/09 4.9/(7-4/ 0.09*6/0* 7! *.-**7-4) 4.!((7-4) 4./ u* 4.*(!7-4) 4.4:0 4.**(4(6( 7* *.!/ 4.4-)0-9* mu* !.(- 4.-*/(*/ 4.!-)7-4) !.944-09 u0 4.**(4(6( 4./ 9.!/ 4.*!/-(4) 4.4-)--!! 4.(- 4.)/!!4!! 4.4! 4.4! 4.!-(9066 !.44!)-4mumax 4.44! 4.(8* 4.(- 4.*(!7-4) !.-0*7-4( 4.9/(7-4/ 4.440!/09 4.**(4(6( 4.-**7-4) *.*44!4*0 4.-4/)946 !.**(4(6( 4.-4)*-!* !.-0*7-4( -.4! 4.e0max 4.-4/)946 4./ 4.-*/(*/ 4.)/!!4!! 4.44.

! 4.!!00009 4.! !4 04 4.4*99(/0 4.*607-4) d5c -4.4*99(/0 -4.)6 0 ! 4.-!0!9(! 0.! 4.! 4.9"la !444 5ostar 4.! 4.! .! 4.4406(/ v! ! ! ! ! v0 4.! 5g4 5s04 . 4.4907-44) 0.9- 0 ! 4.!!!7- 4.4(4(((gam! !4 gam0 !4 !4 !4 !4 !4 !4 !4 d5t 6.9- 0 ! !444 !444 !444 4.)6 phi0 phi* phi) 4.!4(-60* sigmamuv 4.) !4 04 4.6-946*( 4.) !4 04 4.0607-4- !.)6 --.44)4096 4.) v* 4.)6 4.*607-4) 4.alphastar 4.4007-4( phi! 4.9- 0 ! 4.-!4!906 0.44)4096 4.) !4 04 4.6-946*( 9.4(4(((4.9(67-4) 4.

&!966+ shows the sustained oscillations in all . yeast cells can exhibit s#ontaneous metabolic oscillations over a range of operating conditions. as the cells are changing from oxidative to fermentative metabolism.4. The figure from Aorro et al. depending on other culture conditions.. controlled mainly by impeller agitation rates. which are different in these experimental studies. such as oxygen supply rates. 1everal experimentalists have documented these oscillations in aerobic continuous cultures of yeast on glucose.4 Metabolic +scillations : C"bernetic Model 2t the intermediate dilution rates. (hemostat or continuous culture steady state data on cell mass yield on glucose The transition from the oxidative to fermentative pathways occurs either gradually or abruptly.7.Figure . such as the agitation rate or oxygen mass transfer rate. 2.

!/ hr-! and oxygen mass transfer rate -. These spontaneous oscillations change in shape. The simulations were conducted on the Ner"eley Madonna software at the operating conditions of dilution rate of 4. glucose.ml+. .the metabolite concentrations measured in the continuous bioreactor. period and amplitude as the bioreactor operating conditions of dilution rate and agitation rate are varied within their oscillatory ranges. the oscillations die down to either oxidative or fermentative consumption of glucose. The top panel shows the online measurement trace of dissolved oxygen concentration in the culture medium. The parameter values for the different model constants are listed in the Table '. intracellular storage carbohydrates &trehalose and glycogen+.(. the cybernetic model predicts also the experimentally observed trends in the shape and period of oscillations as well as the damping of the oscillations to either the fermentative or the oxidative consumption outside the range of oscillatory conditions. The metabolic oscillations can also be predicted by the yeast cybernetic model equations given above &Kones and :ompala.!.a &strongly affected by the agitation rate+ of *44 hr-!.7 solver. which is most readily obtained. and cell number concentration &L. 2s these operating conditions are varied outside their oscillatory range. The subsequent panels show off-line measurements of ethanol. 2t different operating conditions. !999+ but requires the use of a stiff M. medium pG.).

( 4.!-9!)00 !.)) 4.P+.*4(-9*! e0 !.!4*)-)6 e* !.4! 4.! -*06.!-/!0*9 0.B.6 mu!max 4./) /.!9 4.4! 4.6 !64!.!9 4.*7-4- 4.*7-44.47-4) 4.)) 4.( .0)*! 5g 4.!9 4.47-4) 4.( 4.( 4.( 4.4007-4( beta! gam* 4.E0 3ariable initial value minimal value maximal value final value t 4 4 4.--!/ 5o !.! !64!.--!/ 4.( 4.4*---/- !.47-4) 4.( 4.4007-4( 4.46!/*09 5t 4.! 4.6 4.6 4.47-4) 4.( 4.! -!.! 4.! 0.0 6.0 4.*4((!* !.!!-7@4) 4.4*---/e! 6.-M.!!-7@4) 5e 4.4! beta0 beta* mu0max 4.( 4.( 4.4! 4.0)*! -!.)) 4.! 04 04 5c 4.! -*06.!9 4.( 4.)) :o0 4.( 4.

!4*/49.44! 4.-!)096 9.0 4.06*4!69 e!max 4.*4/9!/ :! 4.4! 4.--440** 4.mu*max 4.* 4.* 4.06*4!69 4.4940/(4.* 4.!/9*4(( !.-**7-4) *.**(4(6( 4.0/*!-(9 e0max 4.69*7-4) mumax 4.*/ 4.06*4!69 4.49/*!( 4.4:0 4.-!)096 mu0 -.4! 4.4- 4.* 0.4(-996) 4.0 4.!-)7-4) *.44! 4.* 4.06*4!69 4.* 4.4! :* 4.* 0.0 4.0-07-4-4.-*69460 9.4607-4-.* 0.* 0.* e*max 4.-*69460 4.4607-44.0 4.**(4(6( 4.*/ alpha* :o* alpha! alpha0 4.**(4(6( 7* *.**(4(6( 4.44! 4.0-07-44.0/*!-(9 4.))-7-4-4.0/*!-(9 4.0-07-4- .* 4.*/ 4.44! mu! 9.)(!))() 4.4! 4.!/66-*9 70 0.9/(7-4) 4.!-)7-4) !.-**7-4) 4.9/(7-4) 0.* 4.44.*/ 4.066)*/* 7! *.44.-(.4)--!*/ mu* ).0/*!-(9 4.

*!(0!-6 ! v0 4.46(()-! 4.(- 4.4/0!96 -!.)!67-49**.)!67-4) 4./ 4.6!-6/00 -4.)!67-4!.!/ 4.466)9/sigmamuv 4./*09) 9**.0(4/0!( -4.!/ 4.4!9!909 4./ 4.-64)6!) 4. 4 4.466)9/- ! !.!(-7-4) -4.46!)!/) 8! 4.! 4.!(4/)(( 4.! 5g4 5s04 .(8* 4.0*/(4*( -4./ 4.-)-(06/ !.(- 4./*09) gam! !4 gam0 !4 !4 !4 !4 !4 !4 !4 d5t -!.! !4 04 4 !4 04 4 !4 04 4 ! !4 04 v* 4.!/ 4.-)90949 4.(- 4.)4-)90) -!.u! 4.)!67-4) d5c !.! 4.!/ 80 4.6*67-4( .*(6/!(9 4.9!999-/ u0 4.9!999-/ 4.44!)!0 alphastar 4.-!6*0( 4.44!-*)6 v! ! ! -!.)90/()/ 4.)64))/) -4./ u* 4.

)6 phi0 phi* phi) 4.! Figure !.9"la *44 5ostar 4. the shape and period of oscillations change as well in both experimental data and model simulations. (ybernetic model simulations of metabolic oscillations in yeast continuous cultures.! 4.)6 4.9- 0 ! *44 *44 *44 4.! 4. .)6 0 ! 4.)6 4.9- 0 ! 4. 2he shape and period of the oscillations in all the metabolite concentrations agree -ualitati)ely with the experimental data.! 4.9- 0 ! 4. /s the bioreactor operating conditions of D and kLa are changed.phi! 4.

7thanol.4.! (onstituti)e Enzyme %ynthesis ate (onstant 4.7.3 9# 92 93 :M0 :M* 8! 80 8* ai a?i bi g! g0 g* f! f0 f* f) 5D.2 mmax.(<ield (oefficient for Ethanol 6xidation 4.g g. and .F g. g:. g.0 4.F g./4 <ield (oefficient for 5lucose 6xidation 4.1.g g.g hr-! hr-! hr-! g. 510 5D."" Fermentation 3aximum %pecific 5rowth ate for Ethanol 1. 57.g g.F g.( Enzyme Degradation ate (onstant !4 ate (onstant for (arbohydrate Degradation !4 ate (onstant for (arbohydrate Degradation 4.38 6xidation 3onod %aturation (onstant for 5lucose 1.1# 3onod %aturation (onstant for 5lucose 6xidation 1..)6 %toichiometric (oefficient for Ethanol 6xidation 0 %toichiometric (oefficient for 5lucose 6xidation ! 4. g:.g g. %pecific 5rowth ate for 5lucose 1. 51!.# mmax.#7 6xidation 3aximum %pecific 5rowth ate for 5lucose 1.6 ate (onstant for (arbohydrate %ynthesis %toichiometric (oefficient for Ethanol +roduction 4.g g.F 3ax.g g.4! 6xygen %aturation (onstant for 5lucose 6xidation 0.8 ?omenclature and Parameter Balues used in Model E9uations and Simulations Aarameter . var.4.11# 6xygen %aturation (onstant for Ethanol 6xidation 4. 5ellmass 5oncentration 1ubstrate 5oncentration Dlucose. 514.g g.issolved Mxygen variable variable variable .g g. Fermentation 3onod %aturation (onstant for Ethanol 6xidation 1.* Enzyme %ynthesis ate (onstant 4.9%toichiometric (oefficient for (arbohydrate +roduction =nlet or Feed 5lucose and %ubstrate (oncentrations!4. 55 51.!/ <ield (oefficient for 5lucose Fermentation 4.nits . 5M hr 4# hr4# hr4# g:.g g.F g./able 2.efinition 3alue mmax.F g.

i 7i mi .C Biotechnolog" and Bioengineering >$:!4))-!4-. 1how that the enzyme modification of the Monod growth "inetics is capable of simulating the presence or absence of the initial lag phase by varying the initial level of the intracellular enzyme content. Kones..1.T.. %ntracellular 7nzyme 5oncentration and its maximum 'elative 2mount of %ntracellular 7nzyme Drowth 'ate on iHth 1ubstrate or Aathway . . Biotechnolog" 780 !4--!*! &!999+.g g. :ompala. )omework Problems Based on this Sample Section 0 !.. Tsao.ilution 'ate 5ybernetic 3ariables 5ontrolling 7nzyme 1ynthesis Mass transfer coefficient for . .issolved Mxygen variable variable variable variable variable variable 5ybernetic 3ariables 5ontrolling 7nzyme 2ctivity variable variable 2eferences0 !.&!96/+. .N.C C... and .. I.concentrations 5T e%. C%nvestigation of bacterial growth on mixed substrates. 0. :ompala. u v "Fa g.1. :.g hr hr hr ! ! ! %ntracellular 1torage 5arbohydrate &Trehalose+ 5onc. C5ybernetic model of the growth dynamics of Saccha"omyces ce"evisiae in batch and continuous cultures. Kansen and D. emax. 'am"rishna. 7xperimental evaluation of cybernetic models.

glucose.*4 hr-!. 85. g glucose. low-high.Q > 4. The Monod growth rate parameters are same as the ones in the problem above.4! g. 1how that the enzyme modification of Monod growth "inetics does not affect the chemostat profiles of cell mass and substrate over the range of dilution rates as well as the washout and optimal dilution rates. and 8p. 8x. Eshe"ichia coli grows on a mixture of three sugars: glucose. :s > 4.F xylose.gP for xylose: mmax.)6 g ethanol . :F > 4. *.F > 4. and 8p.0 hr-!.F glucose.g. Gow will these sugars be consumed in a batch bioreactor with the initial sugar concentrations of . xylose and lactose.s > . 85.6 hr-!.!4 gdw.l.D > 4.F xylose.g. -.! g. /ymomonas mobilis has been engineered to ferment pentoses li"e xylose in addition to the common hexose.F of lactose. :D > 4. :Q > 4..F glucose.-0 g.hr-!.g. and 0.! g. coli is to be cultured in a chemostat on a mixture of glucose.D > !. The Monod growth rate parameters for glucose are: mmax. high-low. 85.F.!! gdw . 7xtend the cybernetic model framewor" to address three substrates.?5c+J /.F > 4.l. g1.F of lactose. and low-low where high represents 99O of emax.i and low represents !O of emax.gP and for lactose: mmax.g.g. 8x.s > 4. #hat is the optimal dilution rate that will maximize the cell mass production rate &.s > 4.g.0. E.4.F. ).). xylose and lactose with the feed concentrations of .i+.g.g.g1. !4 g. The Monod parameters for this metabolically engineered microorganism during growth on glucose are mmax > 4. and 0.s > 4. :s > 4.)4 hr-!.-. The same parameters for growth on xylose are mmax > 4.F.4. 7xamine whether the order of substrate preference is affected by the choice of initial levels of the two "ey combinations &test the four combinations: high-high. !4 g.g.Q > !.

Saccha"omyces ce"evisiae grows in a typical diauxic growth phenomenon on a mixture of glucose and galactose. 0.4.for both "ey enzymes and without using the assumptions stated in the last two sentences of above problem. .l.etermine how the cell mass. g galactose. The Monod growth rate parameters for the fermentative growth on galactose are assumed to be mmax > 4. and 8p. operated at high dilution rate. :s > 4. g galactose.s > 4.F.!. using a > 4. glucose. .s > 4.F galactose.l and that both sugars are fermented at lower dilution rates or glucose concentration R 4.se the growth parameters from the earlier problem statement.g xylose.l xylose. The growth rate parameters for oxidative growth on galactose are assumed to be mmax > 4. if the inoculum is .F glucose and 04 g. Ma"e any other assumptions as needed. 6.F and 8x..g ethanol.(.. 2ssume that the fermentative growth on xylose gets repressed or shut off completely when the glucose concentration exceeds 4. g galactose.** hr-!.-6 g cell mass . 5ontinuing with the theme of above problem. 1imilar parameters for growth on glucose and ethanol are given in Table '. (.).)( g ethanol . galactose and ethanol profiles will be in batch culture on a mixture of !4 g.444! and b > 4.44! g.l glucose and -4 g. .4. #hat dilution rate should be used to maximize the ethanol production rate from the metabolically engineering /ymomonas mobilisJ .). 8x.g cell mass .0 g. These cells are grown in a chemostat fermentor with the feed containing -4 g. to ensure that all of xylose is consumed in continuous culture.F. a second chemostat &of the same size+ is added in series or downstream from the first chemostat.!. 1olve the above problem using the cybernetic model equations.sing the Monod chemostat equations. :M* > 0.s > 4.se the cybernetic model equations or state your additional assumptions.0 g. calculate the maximum ethanol production rate possible from these cells growing in a chemostat.)4 hr-!.g.! g. :s > 4.

a of !444 hr-!J #atch out for the possibility of spontaneous metabolic oscillations.(.). assuming a -. !*.?57+. #hat should be the dilution rate used in the single chemostat.F.?5c+. #hat should be the dilution rate used in the single chemostat. and &b+ Alot how the period changes with the mass transfer coefficient. ce"evisiae is grown in a chemostat on a mixture of glucose and galactose. using the cybernetic model parameters in Kones and :ompala &!999+ &given in the Table '.). -.F. along with glucose feed concentration of *4 g. to maximize the ethanol production rate &.a of !44 hr-!J !0. %nvestigate whether the cybernetic model equations given in section '.) can predict the elimination of oscillations with the inclusion of ethanol in the feed stream. ce"evisiae is grown in a chemostat on a mixture of glucose and galactose. %t is desired to maximize the cell mass production rate &. with the feed concentration of each being -4 g. ce"evisiae can be avoided by adding a small amount of ethanol to the feed stream. 5ontinuing with the theme of the above problem further.F. 11. 11.se a sti!! equation solver in your simulations to obtain the oscillations. 5ontinuing with the theme of the above problem.(. %t has been found experimentally that the spontaneous metabolic oscillations in continuous cultures of yeast S.a. 1imulate the metabolic oscillations of yeast. . #hat is the smallest ethanol concentration that will eliminate the oscillationsJ . S. S. with the feed concentration of each being -4 g. assuming a -.ta"en from continuous cultures on a mixture of glucose and galactose at a dilution rate.!+ to &a+ Alot how the period of oscillations changes with dilution rate .

2 second fermentor will also contain glucose as the sole carbon substrate for the yeast to utilize in the absence of oxygen.a. Iext vary each of the assumed parameters to determine if and how the shape and period of oscillations change from the base case. and the different patterns of multiple substrate utilization in batch cultures.aborator" E. you will be able to analyze the "inetics of cell growth. measure the cell mass concentration &through optical density+ and determine the concentrations of glucose and ethanol with spectrophotometric assay "its. we will study the growth characteristics of the yeast Saccha"omyces ce"evisiae in batch cultures. 2 third lab-scale fermentor will be used to observe the growth behavior when the cells are presented with a mixture of two carbohydrates. . glucose and glycerol in the presence of oxygen. irst obtain the oscillations through numerical simulations of the cybernetic model for any combination of bioreactor operating parameters.(. and -. . 2 labscale &. .ercise0 -east *ermentation 86 =ntroduction %n this laboratory exercise.!). :M* and 8*. mmax. #ith the accumulated results from the all the students over eighteen hours for the three different fermentors. a?. b.etermine the parameter sensitivity of the cybernetic model simulations of spontaneous metabolic oscillations in continuous cultures of yeast for the following assumed parameter values: a. 8ou will ta"e samples from the fermentors.liter+ fermentor will be used to study batch growth "inetics of the yeast growing on glucose as the single carbon substrate provided in the presence of oxygen. :*. .

hr-! with a low biomass yield of 4. The stoichiometry of this reaction is: 5/G!0M/ @ /M0 /5M0 @ /G0M @ e --------------E . 0+ The oxidation of glucose.0.l in aerobic cultures.hr-! with a biomass yield of about 4. and a high energy yield of !/-06 2TA per mole of glucose metabolized.). a respiratory quotient of about !. which occurs primarily when the glucose concentration is high or when oxygen is not available. The cells attain a maximum specific growth rate of only about 4.g dry mass per gram glucose consumed and a high respiratory quotient &the ratio of 5M0 production rate to the M0 consumption rate+ and a low energy yield of only about 0 2TA per mole of glucose metabolized. which predominates at glucose concentrations below -4 mg.g dry mass per gram glucose consumed..!. The cells attain a maximum specific growth rate of about 4. The stoichiometry of this reaction is @ 05M0 @ e 5/G!0M/ --------------E 050G-MG where e represents chemical energy utilized in the growth processes.-east Metabolism Saccha"omyces ce"evisiae uses the following three ma$or pathways for growth on glucose: !+ The fermentation of glucose.

and the impellers inside each fermentor will be operated at )44 rpm. The other batch fermentor will have two initial carbon sources . 2fter the depletion of the faster growth-supporting substrate.perimental Conditions The temperature of the water bath surrounding the fermentors will be controlled at *4S5. glucose.glucose at * g. which predominates when fermentative substrates are not available or in very limited supply. and an exponential growth phase on glycerol is expected to follow a diauxic lag phase. The dissolved oxygen concentration in all the three fermentors can be computer controlled to maintain a desired level by enriching the oxygen concentration in the sparged gas. a low respiratory quotient of about 4. The stoichiometry of this reaction is: 50G-MG @ *M0 05M0 @ *G0M @ e --------------E .tilization of glycerol &the second carbon substrate provided in the third fermentor+ by Saccha"omyces ce"evisiae is repressed by glucose. 2ir will be sparged into the first and the third fermentors at constant rate of !4 l. The cells attain a maximum specific growth rate of about 4. E.l.ab Procedures . the enzymes necessary for the assimilation of glycerol is induced.*+The oxidation of ethanol. =ntroduction to ./-4.0 hr-! with a high biomass yield of about 4.l. and an energy yield of about /-!! 2TA per mole of ethanol metabolized. Two of the three fermentors will have glucose as the only initial carbon source at a concentration of !4 g.( g dry mass per gram ethanol consumed.(.l and glycerol at ( g.min.

1et the wavelength to /*4 nm. The calibration curve correlating cell concentration with absorbance deviates from a linear correlation at high cell densities. and assayed for glucose and ethanol by the spectrophotometric assay.6 Determining Cell Concentration !+ Teroing the spectrophotometer. itHs a good idea to dilute any of your high M. .8ou will monitor the growth characteristics of the yeast in the batch fermentors in three ways by measuring the concentration of cells. To remove the cells from a * ml sample. values fall on the linear portion. 2) Experimental Procedures . &glycerol+ and ethanol in the cell-free culture samples will be analyzed by high performance liquid chromatography or spectrophotometric assay. the cell suspension will be centrifuged and the supernatant will be filtered through a microfiltration syringe.sing the "nob on the left.l+ in the sample is proportional to the absorbance reading on the spectrophotometer. The concentration of glucose. set the reading to 4O transmission when the chamber is emptyP using the "nob on the right. samples &that may be on the non-linear portion of the curve+ by a "nown dilution factor to confirm that the measured M. 8ou will ta"e a sample of the culture medium from the fermentor and read its absorbance using a spectrophotometer. . and preparing cell-free samples at hourly intervals from each bioreactor for assaying the concentrations of glucose and ethanol. 8east cell concentration can be determined indirectly by measuring the optical density &absorbance+ of a culture sample. set the reading to !44O transmission when the . the concentration of yeast cells &gdw. Necause of this.p to a certain cell density.

raw a graph of: a+ Fogarithm of cell concentration vs. time . 5lean the outside of the test tube with ethanol. and record the absorbance reading.0-. Ta"e another 6-!4 ml sample from your groupHs fermentor and gently mix. the culture will be centrifuged and the supernatant will be filtered through a microfiltration syringe. 'ecord the time you ta"e the sample along with the absorbance reading in the linear range as well as the dilution factor. 0+ . %f the absorbance reading is greater than 4. &6 2eport : Due *ebruar" 8'# 8''7 2+ . B6 Spectrophotometric . time b+ Dlucose concentration vs. irst flush out the sample tube for your groupHs fermentor by ta"ing an 6-!4 ml sample. Ta"e about ) ml from your sample tube and transfer it to a glass test tube. and measure the absorbance again to chec" if the absorbance reading is on the linear portion of the calibration curve. insert it into the spectrophotometer. a typical limit of linear correlation between the absorbance and cell mass concentration. The lab T2 will provide more details on the assay procedures during the experiment. dilute the sample with a "nown amount of pure medium.chamber contains a test tube with about ) ml of pure medium.ssa"s for Glucose and Ethanol To remove the cells from a * ml sample. which you will then discard. time c+ 7thanol concentration vs.etermining cell concentration.

&glycerol+ and ethanol concentration profiles loo" as they do for each batch fermentor. 1pecifically. N+ . glucose.for each of the three batch fermentors.etermine the specific growth rate and the yield coefficient &gram dry weight of cells produced per gram of carbon source consumed+ for each growth phase in the three fermentors. 5+ %nterpret these two graphs in light of the bac"ground information on yeast metabolic pathways and the CcyberneticC principle that cells choose to grow at the fastest possible rate. ogler U Durmen V 0446 . &The calibration between the absorbance reading and the dry cell mass concentration of the yeast cells will be performed at the end of the batch cultures and provided in the following class period+.niversity of Michigan . discuss why the cell mass.

- Application of Chemical Engineering Principles to the Tasks and Activities for Industrial Internship Accreditation
- Your First Microbiology Experiment
- Science Demonstration
- Yeast Medium Modification
- Yeast in Batch Culture
- Assymetric Reduction of OIP to DOIP_using Shake Flask_part 1_1st Ed
- Tutorial 6-Oxygen Transfer in Reactors
- Bioseparations in Two-Phase Aqueous Micellar Systems
- Ethanol Assay
- Algorithm 3
- Yeast in Batch Culture_Expt_6_part 2
- Assymetric Reduction of OIP to DOIP_using Shake Flask_part 1_1st Ed
- MEP201 Fourier Series 2006
- Lecture 6 - Metabolic Modeling
- Legal Memo
- Yeast in Batch Culture_Expt_6_part 2
- Yeast in Batch Culture_Expt_4_shake Flask_part 1_4th Ed
- Effect of Dissolved Oxygen, Temperature, Initial Cell Count
- Variation in the Analysis of Real ChE Process
- Technical Writing in Chemical Engineering
- Lecture 4 - Physical Bioprocesses 2
- The chemistry of casks – Part 2 _ Production _ bourbon, chemistry, cognac, Oak maturation, rum, whiskey, whisky
- The chemistry of casks – Part 3_ Wood chips vs
- Solved ProblemSetEquipDesign2

Data

Data

- Co-Production of Bioethanol, Lactic Acid, Electricity and Heat From Lignocellulosic Biomass
- Mixture Design as First Step for Improved Glutaminase
- Long Resume
- BChE Questions Syllabus
- Glycolysis Lecture
- benzaldehyde.l-pac
- Enzymes aEnzymes and Their Functions
- Historia de La Biotecnologia-1
- Dr Najeeb Glycolysis
- Effect of Different Carbon Sources on the Production of Succinic Acid Using Metabolically Engineered Escherichia Coli
- High Consistency Hyrolysis
- Differential media lab
- Fermentation
- 1-s2.0-S0141022999001556-main
- 12.Tay Et Al-2002-Biotechnology and Bioengineering
- Bich411 Enzyme Regulation.pdf
- Effect of pH on Bacillus Thermoamylovorans Growth And
- Lec 20
- wright_etal_systems analysis of TCA cycle2.pdf
- Influence of Glucose on Glycerol Metabolism
- journal
- Production of Pyruvate and Acetaldehde
- Biochem Samples Test5
- Modeling Metabolic Path
- Biotechnological Applications of Industrially Important Amylase Ebzyme
- Far Men Tat Ion
- Process Description
- bi00480a010
- Fermentation & Downstream
- Study the Rates of Fermentation of Fruit or Vegetable Juices
- Microbial Growth on Multiple Substrates

Are you sure?

This action might not be possible to undo. Are you sure you want to continue?

We've moved you to where you read on your other device.

Get the full title to continue

Get the full title to continue reading from where you left off, or restart the preview.

scribd