Batch cultivation tutorial
Worked Problems
#1. A batch reactor is inoculated with 0.5g/L of bacteria. The specific growth rate (µ)
is 0.6 h-1. Assuming there is no lag phase and substrate is unlimited, what will the
biomass concentration be after 6 hours?
Given:
Xo=0.5 g/L
µ=0.6 h-1
t= 6h
Required, X?
Solution
• Since this is the exponential growth we use :
𝑋
ln = µ𝑡
𝑋𝑜
X= 𝑋𝑜 𝑒 −µ𝑡
X= 0.5 × 𝑒 (0.6×6)
X= 0.5 × 36.6 = 18.3 𝑔/𝐿
#2. A fermentation process uses glucose as the limiting substrate. The
initial glucose concentration S0 is 20 g/L. The inoculum, Xo is 1.0 g/L. if
the yield coefficient is 0.4 g biomass/g glucose, calculate the maximum
biomass concentration at the end of one batch
Given:
So= 20 g/L
Xo= 1.0 g/L
YX/S= 0.4 g/g
Required: Xmax?
Solution
• Maximum biomass (Xmax) is reached when the limiting substrate is
completely depleted. Therefore, final substrate Sf=0
𝑋𝑚𝑎𝑥 = 𝑋𝑜 + 𝑌𝑋/𝑆 (𝑆𝑜 − 𝑆𝑓 )
𝑋𝑚𝑎𝑥 = 1 + 0.4 (20 − 0)
𝑋𝑚𝑎𝑥 = 1 + 8.0
𝑋𝑚𝑎𝑥 = 9.0 𝑔/𝐿
*In reality the substrate might not go exactly to zero due to Ks limits , for
yield calculation, complete consumption is a standard assumption
#3. A strain of mold was grown in a batch culture on glucose and the following
data were obtained.
Ideal CSTR
• Ideal CSTR (chemostat) allows for the
continuous culture of M.O
Key Mechanisms
• Fresh sterile medium containing a
limiting nutrient, is fed into a reactor
at constant rate, while culture liquid
containing cells and metabolic
products is removed at the same rate
General Mass balance
• The fundamental equation for any component in the reactor is derived
from the law of conservation of mass
Accumulation= 𝐼𝑛 − 𝑜𝑢𝑡 + 𝑔𝑒𝑛𝑟𝑎𝑡𝑖𝑜𝑛 − 𝐶𝑜𝑛𝑠𝑢𝑚𝑝𝑡𝑖𝑜𝑛
𝑑𝑉𝐶 Eq.1
= 𝐹𝐶0 − 𝐹𝐶 + 𝑟𝐶 𝑉
𝑑𝑡
Where, C= concentration (cells, substrate)
F= volumetric flowrate
V=reactor volume
rC= reaction rate
Cell mass balance and dilution rate
At steady state, dX/dt=0, With sterile feed; X0=0
0= −𝐹𝑋 + µ𝑋𝑉 Eq.2
𝐹
µ= =𝐷
𝑉 Eq.3
The dilution rate(D)
• The dilution rate, D is the reciprocal of the residence time. It represents how many
reactor volume pass through per unit time
* At a steady state in a chemostat, the specific growth rate of the cells (µ), is
controlled by the dilution rate (D)
Monod’s growth kinetics
• The relationship between specific growth
rateµ and substrate concentration is modeled
by Monod’s equation
µ𝑚𝑎𝑥 𝑆
µ= Eq.4
𝐾𝑠 + 𝑆
Where µmax: maximum specific growth rate
KS: half velocity constant (Value of S, where
µ=1/2 µmax
We may also linearize Eq.4. for simplicity * For a graph of 1/ µ vs 1/ X, KS/ µ max
is the slope
1 𝐾𝑠 1 1
= +
µ µ𝑚𝑎𝑥 𝑆 µ𝑚𝑎𝑥 Eq.5
Substrate mass balance
• We apply the general mass balance for the limiting substrate (S),
• Yield coefficient (YX/S)
1 µ𝑋 Eq.6
𝑟𝑠 = − 𝑟𝑥 = −
𝑌𝑋/𝑆 𝑌𝑋/𝑆
Steady state equation Accumulation, (dS/dt)=0
µ𝑋𝑉
𝐹𝑆𝑜 − 𝐹𝑆 − =0 Eq.7
𝑌𝑋/𝑆
Dividing by V (where D=F/V) and substituting µ = 𝐷
𝐷𝑋 Eq.8
𝐷𝑆𝑜 − 𝐷𝑆 − =0
𝑌𝑋/𝑆
Steady state substrate concentration
• Since µ=D at steady state, we can rearrange to solve Monod’s equation (E.q.4) for the
steady state substrate concentration.
𝐾𝑆 𝐷
S= Eq.9
µ𝑚𝑎𝑥 − 𝐷
This equation shows that the residual substrate concentration is determined solely by
the dilution rate independent of the inlet feed concentration S0
Steady state biomass
concentration
• Rearranging eq.8, the steady state biomass concentration will be
𝑋 = 𝑌𝑋/𝑆 (𝑆0 − 𝑆) Eq.10
Substituting for S from eq. 9 in to Eq.10
𝐾𝑠 𝐷
𝑋 = 𝑌𝑋/𝑆 (𝑆0 − ) Eq.11
µ𝑚𝑎𝑥 − 𝐷
Chemostat dynamics
• Low D: Substrate is nearly fully consumed,
biomass (X) is at its maximum
• Increasing D: substrate S rises, because
cells have less residence time to consume
it
• As D approaches µmax, X drops to Zero and
S approaches S0
Product formation Kinetics
Growth Associated products
For products directly linked to growth (e.g. ethanol in anaerobic yeast fermentation),
the rate of product formation is proportional to the growth rate
𝑟𝑃 = 𝑌𝑃/𝑋 𝑟𝑋
Eq. 12
Steady state product concentration P, becomes
P= 𝑌𝑝/𝑋 𝑋 Eq. 13
Washout condition
• Washout occurs when the dilution rate exceeds the maximum possible growth rate of
M.O.
• At this point, cells are removed from the reactor faster than they can reproduce.
Biomass concentration (X) drops to zero
Critical dilution rate
This is a condition where steady state S, approaches the inlet concentration S0
µ𝑚𝑎𝑥 𝑆0
𝐷𝑐𝑟𝑖𝑡 = Eq. 14
𝐾𝑠 + 𝑆0
Note: if S0>> KS then Dcrit=µmax
Biomass productivity
• Productivity (PD) Is the rate of biomass
production per unit reactor volume
PD= 𝐷 × 𝑋 Eq. 15
The graph shows a parabolic shape. At
low D, X is high but flow is low. At high
D, flow is high but X is low (washout).
The maximum occurs at Dopt
Optimum dilution rate (Dopt)
• To find the dilution rate that maximizes productivity, we differentiate DX with
respect to D and set it to zero
𝑑(𝐷𝑋)
=0 Eq. 16
𝑑𝐷
The solution is
𝐾𝑠
𝐷𝑜𝑝𝑡 = µ𝑚𝑎𝑥 (1 − √ ) Eq. 17
𝐾𝑠 + 𝑆0
Example
• A chemostat study was performed with yeast. The medium flow rate was varied and
the steady-state concentration of cells and glucose in the fermenter were measured
and recorded. The inlet concentration of glucose was set at 100 g/L. The volume of the
fermenter contents was 500 mL. The inlet stream was sterile.
a. Find the rate equation for cell growth.
b. What should be the range of the flow rate to prevent washout of the cells?
Solution: a. Let’s assume that the growth rate can be expressed by Monod kinetics. If
this assumption is reasonable, the plot of 1/µ versus 1/X will result in a straight according
to Eq.5. At steady state the dilution rate D=µ. The dilution rate for the chemostat is,
D=F/V;
1 𝐾𝑠 1 1
= +
µ µ𝑚𝑎𝑥 𝑆 µ𝑚𝑎𝑥
The plot of 1/D vs 1/S is shown in the figure; a
straight line with intercept
1 𝐾𝑆
= 3.8 And the slope, = 5.2
µ𝑚𝑎𝑥 µ𝑚𝑎𝑥
Therefore, µmax=0.26 hr-1, and KS =1.37 g/L. the rate
equation of cell growth
µ𝑚𝑎𝑥 𝑆𝑋 0.26 × 𝑆 × 𝑋
𝑟𝑋 = µ × 𝑋 = 𝑟𝑋 =
𝐾𝑆 + 𝑆 1.37 + 𝑆
b. To prevent washout of the cells, the dilution rate should
be less than D critical. Therefore, from Eq. (14)
µ𝑚𝑎𝑥 𝑆0 0.26 × 100
𝐷𝑐𝑟𝑖𝑡 = = = 0.25
𝐾𝑠 + 𝑆0 1.37 + 100
𝐹
D= Rearranging F = 0.25 × 0.5 = 0.12 𝐿/ℎ
𝑉
* To prevent washout the flow should not be more than 0.12 L/h