Lab 1.1
Lab 1.1
Introduction
In a batch culture, there is neither input fed nor output produced during the fermentation
process. Initially, the medium culture is inoculated with the microorganism. The growth
constantly rises until at certain level, where it is inhibited because of the dropping substrate
concentration and the presence of toxic metabolites. During the growth of the cells in a batch
fermentation, they follow a growth curve analogous to the one shown in Figure 1. The growth
curve contains four distinct regions known as phases. They are known as Lag Phase, Log
Phase, Stationary Phase, and Death Phase.
Lag phase occurs between inoculation and the maximum growth rate. It is the first major
phase of microbial growth in a batch fermentation process. It is also a phase of adaptation of
the cells to their new environment. During this phase, there is only a minimal increase in cell
density. In certain types of fermentation, this phase may not even happened at all. [1]
There
are two sub phases in the lag phase. In the first phase, there is no growth acknowledged while
in the second sub phase, also known as acceleration phase, a constant growth begins.
The second phase is the exponential phase. The cells begin to multiply with their maximum
growth rate. This phase is crucial to determine the maximum growth rate, μ max and doubling
time, d since the growth at this time is the most constant and ideal. This is the second major
phase in a batch fermentation process. It is also known as the logarithmic growth phase.
During this phase, cells have adjusted to their new environment. They are dividing at a
constant rate resulting in an exponential increase in the number of cells present. This is
known as the specific growth rate and can be shown mathematically by the first order kinetics
as the following:
dX
=( µ−k d ) X
dt
For (µ−k d ), it can be substitute as µnet . The cell death rate may be negligible only if it is
significantly smaller than the cell growth rate. Next, the growth curve during this phase can
be projected using the Monod batch kinetics:
µmax S
µ=
Ks+S
After retardation phase, the growth phase arrives at the stationary phase which is the third
Figure 2.2: A typical Monod batch kinetics graph
major phase in a batch fermentation process. During this phase, the growth becomes constant
for a period of time before it decreases. This is due to the number of cells that are multiplying
and dying at the same rate. The cells’ death may be due to several factors such as the lack of
growth nutrients, excess of toxic substances, and stress due to recombinant gene. Primary
metabolite or growth associated production cease to exist. While secondary metabolite or
non-growth associated production may continue to run.
Finally, the fourth major phase is the death phase or decline phase. The growth declines from
its stationary phase due to the cells lysis. This is shown by the decrease amount of the viable
cell. In this phase, the cells’ death rate is greater than the cells’ multiplication rate. It can be
shown using the first order kinetics, similar to the exponential phase:
dX
=−k d X
dt
Procedure
Figure 3: The Piping & Instrumentation Design (P&ID) of the batch vessel fermenter
The unit procedure chosen is a batch vessel procedure in a fermenter. The fermenter itself is
labelled as P-1/FR-101 with a vessel volume of 13249.57L. In this simulation laboratory,
only one fermenter was used. The unit cost is $965,000. The major equipment specification
& FOB cost which includes both the fermenter and unlisted equipment is $1,206,000. The
Total Plant Direct Cost (TPDC) is $3,884,000 and the Total Plant Indirect Cost (TPIC) is
$2,330,000. The Total Plant Cost (TPC) which is the sum of TPDC & TPIC is $6,215,000.
The Contractor’s Fee & Contingency (CFC) is 932,000. The Direct Fixed Capital Cost (DFC)
which is the sum of TPC & CFC is $ 7,147,000. The annual labour cost for an operator is
$1,129,801 with an annual amount of 16,374 hours. The annual utilities cost is $37,561
which includes Standard Power, Steam and Glycol. The annual operating cost which includes
labour-dependent, facility-dependent, laboratory, and utilities is $2,684,000.
In this simulation laboratory, the annual operating time is calculated as 7,911.28 hours with a
recipe batch/cycle time of 33.81 hours. The number of batches per year is 234 batches. The
total mass of materials which includes air, biomass, media, and water is 20,194.75 kg/batch
or 4,725,571 kg/year. For the input, media & biomass is 500 kg/batch, air is 9694.75
kg/batch, water is 10,000 kg/batch. The total input is 20,194.75 kg/batch. For the output, the
carbon dioxide is 9,707.34 kg/batch, biomass is 176 kg/batch, impurity is 45 kg/batch, water
is 10,270 kg/batch, and the main bio-product monoclonal antibody (MAB) is 9 kg/batch. The
total output is 20,207.34 kg/batch. Inside the fermenter FR-101, the operation starts with the
operation CHARGE-1 at 0.00 hour and ends at 1.76 hour. Next, the operation CHARGE-2
starts at 1.76 hour and ends at 2.93 hour. Right after that, operation HEAT-1 starts at 2.93
hour and ends at 3.41 hour. Next, the actual fermentation process, operation FERMENT-1
starts at 3.41 hour and ends at 27.41 hour. After that, the operation COOL-1 starts at 27.41
hour and ends at 28.56 hour. Finally, the operation TRANSFER-OUT-1 occurs which starts
at 28.56 hour and ends at 33.81 hour. It also marks the end time for the fermenter.
For one batch cycle, the mass of bio-product monoclonal antibody (MAB) produced is 9.00
kg and the Carbon Dioxide vented is 314.90 kg. The media was converted 100% without
leaving any amount at the output stream. Meanwhile for biomass, it increases to a final mass
of 176.00 kg/batch at the output.
Section Starting Active Amount Molar Mass Yield Gross Mass Yield
Material Product Needed (kg Yield (%) (%)
Sin/kg MP) (%)
Main (none) (none) Unknown Unknown Unknown Unknown
section
Sin = Section Starting Material, Aout = Section Active Product
Media
3. STREAM DETAILS
1. Calculate the amount of air to be supplied for the aerobic fermentation process.
For one batch process, the amount of air supplied for the aerobic fermentation process is
9694.74974 kg.
2. Determine the size of the fermenter necessary for this batch process.
The fermenter size must be at least 13249.57 L of volume.
3. What is the composition of vent & how much CO2 is produced in the batch process?
The composition of vent is as below:
16
14
Biomass Concentration (g/L)
12
10
0
0 5 10 15 20 25
Time (h)
0.8
0.7
MAB Concentration (g/L)
0.6
0.5
0.4
0.3
0.2
0.1
0
0 5 10 15 20 25
Time (h)
40
35
Media Concentration (g/L)
30
25
20
15
10
0
0 5 10 15 20 25
Time (h)