RATE LAWS, STIOCHIOMETRY, MASS BALANCE, DESIGN EQUATIONS

AND PROFILES FOR BIOREACTORS

Bioreactors are classified as:

(A) (B)
Batch Bioreactor Continuous Bioreactor (STR); Chemostats; (Turbidostat)
Batch Bioreactor

(1) RATE LAWS:

Cell Growth Rate
Monod Equation (1) rg = µCc
Monod Equation (2) rg = µmax (CcCs/KsCs)
Monod Equation (3) rg = Kobs (µmax CcCs/KsCs)
Where:
µ = µmax (Cs/ KsCs)
kobs = (1- Cp/Cp*)r

Cell Death Rate

(4) rd = (Kd+KtCt)Cc
Note:
For cell growth rate (rg); following equations can be further used:
a) Tessier Equation:
rg = µmax [1-exp(-Cs/K)]Cs
b) Moser Equation:
rg = µmax Cc/(1 + KCs)
For glucose to ethanol fermentation:-
n = 0.5
Cp* = 93 g/l

Nomenclature:

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rg = cell growth rate (.g/ls..)
µ = specific growth rate (1/s)
Cc = cell concentration (g/l)

µmax = maximum specific growth rate (1/s)

kobs = specific growth rate constant expediently found (1/s)

Cs = substate concentration (g/c)

Cp = product concentraton at any time (g/l)

Cp* = product Concentration at which all metabolism ceases (g/l)

n = empirical constant

Ks = Monod constant (g/l)

r & K = empirical constate fro moser equation

Ct = conc. of a substate toxic to he cell growth (t = texicity)

Kd = natural death rate of cell (death rate const.) (0.1h – 0.0005 h-1)range

Kt = cell death rate due to toxic substate (specific death rate constant)

rd = cell death reate (g/ls)

(2) STIOCHIOMETRY:

(1) rsm = mCc

(2) -rs = Ys/c rg + mCc
(3) rp = rg Yp/c
(4) rp = KpCsnCc/Ksn+Csn

(5) -rsn = mCc + Ysn/p KpCsnCc/Ksn+Csn

Nomenclature:
rsm = rate of substrate consumption for maintenance

Ys/c = fractional yields of substrate in terms of cell growth

Yp/c = fractional yield of product in terms of cell growth

(-rs) = rate of substrate disappearance

rp = product formation in the stationary phase (for eq. 3)

rp = product formation in the stationary phase (for eq. 4)

-rsn = net.rate of substrate consumption during the stationary phase in the

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 presence of Zn dry nutrients
Csn = conc. of secondary nutrient (g/c)

Ksn = constant (in the presence of secondary nutrient) (g/l)

Kp = specific rate constant with respect to product (l/gs)

Before applying mass balance on batch reactor 2 basic equations must be kept
in mind:

(a) For the calculation of nutrient consumption (maybe primary or

secondary or tertiary and maybe single or multiple

Rate of nutrient consumption can be calculated as:

Net rate of substrate Rate consumed by Rate consumed Rate consumed for
consumption cells to form product maintenance
-rs = Ys/c rg + Ys/p rp + mCc

Nutrient here is substrate, it can be accounted in two ways:-

(1) either by counting

(2) or by mass calculations

In microbiology counting will be preferred and in bio chemical engineering

mass calculation will be preferred.

(b) In the stationary phase the conc. of live cells is constant and we will
apply two limiting assumptions
(i) product fraction is only during growth phase
(ii) and product formation only during stationary phase

The product conc. and substrate conc. can be related as

Cp = YP/S (Cso + - Cs)

(3) MASS BALANCE:-

1. Micro-organism or cell balance

Rate of assumption of Rate of cells entering Rate of cells Net rate of

cells (g/s) leaving (g/s) generation of live
calls
V dCc/dt = VoCco - VCc + (rg-rd)V

For batch reactor; Vo and V=o

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 V dCc/dt = (rg-rd) v
Divided by v
dCc/dt = rg-rd …………………………………..(i)

2. Substrate Balance (in the cell growth phase)

Rate of assumption of Rate of substrate Rate of Rate of substrate

substrate (g/s) entering substrate generation (g/s)
(g/s) leaving (g/s)
VdCc/dt = VoCso - VCc + Vsv

Modify for batch rection; Vo and V=o and divided by v.

dCc/dt = Ys/c (-rg)-mCc …………..(ii)

3. Substrate Balance (in stationary phase)

VdCc/dt = -nCcV - Ys/p (-rp)V (7-121)

Dividing by V
dCc/dt = Ys/p (-rp) - mCc …………(iii)

4. Product Balance

VdCp/dt = rpV = Yp/s (-r s)V (7-122)

Dividing by V
dCp/dt = Yp/s (-r s) ………………(iv)

(4) Design Equations:-

Equations i, ii, ii, and iv in the mass balance are called the design equations for
the batch bio reactors. By using these equations and solving them
mathematically or statistically or graphically or using any computer program,
we can find the design parameters like time and different concentrations.

(5) Profiles for Batch Bio-Reactors:-

These profiles may be of following 4 types.

i) Time vs. Conc.

ii) Time vs. Rate

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iii) Time vs. Kobs (see figure at page 404)

iv) Time vs. Cell Growth (cell conc.)

Since fermentation reactions are major two types that is

 Enzymation
 Mocro-bial

The above 5 steps have been developed for batch bio-reactors for micro-bial
fermentation but the same maybe used in the design for the enzymatic
fermentation reactions also by skipping cell growth rate or cell conc. steps and
adding the steps for enzyme consumption during the reaction and its recovery
at the end.

RATE LAWS, STIOCHIOMETRY, MASS BALANCE, DESIGN EQUATIONS

AND PROFILES FOR CONTINUOUS BIOREACTOR (CHEMOSTATS /
TURBIDOSTATE)

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DESIGN EQUATION:

In these design equations three important parameters like max. rate of cell
production (DCc), Concentration of cell (Cc) and conc. of substrate (CS) will be
related as a function of dilution rate (D).

D = ro/v (only used in continuous reactor)

CSTR MASS BALANCES:

Cell: DCc/dt = 0 –DCc + (rg-rd)  (1)

Substrate: dCs/dt = DCso – DCs + rs  (2)

Using monod equation, the growth rate is determined

Rg =µCc  (3) * [µ (specific velocity) = rg/Cc]

µ = µmax + Cs/Ks + Cs  (4)

For steady state operation, we have; dCc/dt=0 and dCs/td=0

DCc = rg – rd  (5)

D (Cso-Cs) = rs  (6)

Combine equation (3) and (5) for steady state operation to obtain the mass flow
rate of cells out of the system moc

moc = Cc ro = rgV = µCcV  (7)

After we divide by CcV

D=µ
[From Equation # 1
DCc = rg-rd
Dcc = rg
DCc= µCc
D = µ]

Inspection of Equation (8) reveals that the specific growth rate of cells can be
controlled by operator, by controlling the dilution rate “D”.

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Using Equation (4) to substitute for “µ” in terms of substrate concentration and
solving for steady state substrate concentration (C/S).

µ = µ max. (Cs/Ks+Cs)

Since µ = D so D = µma*Cs/Ks+Cs

D (Ks + Cs) = µma * Cs

DKs + DCs = µma * Cs

DKs = µmax Cs – DCs

DKs = Cs (µmax – D)

Cs = DKs / µmax – D  (9)

Assuming that a single nutrient is limiting cell growth is the only process
contributing to substrate utilization and that cell maintenance can be
neglected the stoichiometry

- rs = rg Ys/c + rosm  (10)

- rs = rg Ys/c  (11)

Cc = Yc/s (Cso – Cs)  (12)

Using the value of Cs from equation (9) and put in (12)

Cc = Yc/s (Cso – DKs/µmax-D)  (13)

After solving we get

Cc = Yc/s (Cso + Ks)/ µmax-D (µmax Cso / Ks + Cso –D )  (14)

WASH OUT

To learn the effect of increasing the dilution rate, we combine equation (1) and
(3) and set rd = 0 to get

From equation (1)

dCc/dt = - DCc + (rg – rod)

dCc/dt = - DCc + rg

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as we know from equation (3) rg = µCc

DCc/dt = -DCc + µCc = Cc (-D + µ)

dCc/dt = Cc (µ - D)  (15)

We see that if D > µ, then dCc/dt will be negative and cell concentration will
continue to decrease until we reach a point where all cells will be washed out
and Cc = 0

The dilution rate at which wash out will occur is obtained from equation (14)
by setting Cc= 0

Dmax = µma Cso/Ks + Cso 

Flow rate at which wash out occurs

We next want to determine the other extreme conditions for the dilution rate,
which is the rate of maximum cell production. The cell production rate per unit
volume of reactor is the mass flow rate of cells out of the reactor i.e. m oc =CcVo

moc =CcVo

divided by “V”

m/v = Cc Vo/V

µCcV/V = CcVo/V

DCc = VoCc/V  (17)

Substituting for Cc yields; [from (13)]

DCc = DYc/s [ Cso – DKs /µmax – D]  (18)

Figure shows production rat (DCc), cell concentration (Cc) and substrate conc
(Cs) as a function of dilution rate “D” i.e.

(Cc,Cc,Dcc) = f(D)

We observe a maximum in the production rate and this maximum can be

found by differentiating production rate with respect to the dilution rate D; and
put d(DCc/dD) – 0  (19)

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On simplification and differentiation equation (18) we get

Dmaxprod = µ max [ 1-(Ks/Ks+Csc)]  (20)

Dmaxprod found in a equation (20) is infact “D” optimized.

= DCc = DYc/s [ Cs – DKs / µmax-D]

= Taking derivative with respect to D and put it zero.

= d DCc/Dd =O= d/Dd [DYc/s ( Cso – DKs / µmax-D]

= d/Dd (Dd [DYc/s Cso) – d/Dd ( D2Ks Yc/s / µmax-D)

= Cso Yc/s – [(µamx-D) d/D (D2ks Yc/s) -D2Ks Yc/s d/dD/ (µmx-D) / (µmax-D)2 ]

= Cso Yc/s – [(µamx-D) (2DKs Yc/s) - D2Ks Yc/s (o-1) / (µmax-D)2 ]

= Cso Yc/s – [(2DKs Yc/s (µmax-D)- + D2Ks Yc/s (o-1) / (µmax-D)2 ]

= Cso Yc/s – [DKs Yc/s (2(µmax-D)- + D / (µmax-D)2 ]

= Cso Yc/s – [DKs Yc/s (2(µmax-2D+D) / (µmax-D)2 ]

= Cso Yc/s – [DKs Yc/s (2 µmax-D) / (µmax-D)2 ]

= Yc/s [Cso – DKs ((2 µmax-D) / (µmax-D)2 ]

= Cso – DKs (2 µmax-D) / (µmax-D)2

= Cso – (µmax-D)2 / DKs (2 µmax-D) / µamx-D)2

= Cso – (µ 2max - D2- 2D µmax) – 2DKs µmax + D2Ks

= Cso – µ max + Cso D2 - 2D Cso – µmax – 2DKs µmax + D2Ks

2

= Cso – µ max + Cso D2 + D2Ks – 2D Cso µmax – 2DKs µmax

2

= Cso – µ max + D2 (Cso + Ks) – 2D µmax (Cso + Ks)

2

- Cso – µ max + D2 (Cso + Ks) – 2D µmax (Cso + Ks)

2

- Cso –µ max + D2 (Cso + Ks) (D2- 2D µmax)

2

- Cso µmax2 / Cso + Ks = D2- 2D µmax

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Adding & subtracting µ2max

- Cso µmax2 / Cso + Ks = D2- 2D µmax + µmax2 - µmax2

- Cso µmax2 / Cso + Ks = (µmax – D)2 µ2max

µmax2 (-Cso µmax2 / Cso + Ks) = (µmax – D)2

µmax2 (-Cso µmax2 – Cso µmax2 / Cso + Ks) = (µmax – D)2

µmax2Cso + µmax2 Ks - Cso µmax2 / Cso + Ks = (µmax – D)2

µmax2 Ks/ Cso + Ks = (µmax – D)2

µmax2 Ks/ Cso + Ks = (µmax – D)

µmax Ks/ Cso + Ks = µmax – D

D = µmax -µmax Ks/ Cso + Ks

Dmax = µmax [ 1 - Ks/ Cso + Ks

BIOCHEMICAL ENGINEERING

RATE LAWS:

rg = µCc

µ = µmax (Cs / Ks + Cs)

1. rg = µmax (Cc Cs / Ks + Cs) ……….. (7-103)

2. rg = kobs [µmax (Cc Cs / Ks + Cs)]

Kobs = (1- Cp / Cp*)n {n = 0.5; Cp* = 938/3dm}
Glucose to ethanol fermentation
Monod
(i) Tessier Equation ……….. (7-106)
(ii) Moser Equation ……….. (7-107)

3. rd – (hd + htCt) Cc

STIOCHIOMETRY:

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On batch and chemostate (CSTR)

1. rsm ……….. (7-110)

2. -rs ……….. (7-112)

3. rp ……….. (7-113)*

4. rp ……….. (7-114)*

5. -rsn ……….. (7-115)

Rate of nutrient consumption (-rs);

Basic equation or fermentation (7-111)

MASS BALANCE AS BATCH (BIOREACTOR) REACTOR:

7-117 (modify for batch)

7-118 (modify for batch)

(1) 7-119

(2) 7-120

(3) 7-121

(4) 7-122

 Profiles; Figure (E7-9.1); (P # 404)

 Figure; Figure 7-13 (P # 395)

MASS BALANCE ON CSTR (CHEMOSTATS)

Figure (7-15); (P # 401)

Figure (7-16); (P # 405)

D = Vo/V (1/ µ); Dilution Rate;

Prove that D = µ (specific growth rate; S)

7–123; 7-174

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7–125; 7-126

7-127 (mc = ______)

Cs ______ (7-129/7-132)

Wash out; Dmax = ________ (7-134)

DCc= ? Max rate of cell production (DCc)

Find d(Dc)/dt = 0

Dmax production = ________ (7-136)

Max rate of cell production

Profiles; Figure (7-17)

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