1. PROCESS MODELLING AND SIMULATION Biochemical Case Study
PTT303 /2
PROCESS MODELLING AND SIMULATION
SEM 1 (2013/2014)
Biochemical Case Study

2. Student should be able to;
Develop chromatographic process to achieve the desired final product purity, combination with membrane filtration to exchange buffers and concentrate the dilute product solutions.
Create the process and identify uncertainty problem that might happen during the operations.

3. OUTLINE FOR BIOCHEMICAL CASE STUDY
A sequential modular approach to solve for a
moderate complex flowsheet
Some common unit operations in biochemical
industries:
Fermenter
Disk-stack centrifugation
Diafiltration
Chromatography

4. Sequential modular approach
Part 1
Fermentation
Section
Part 3
Purification
Section 2
Sequential modular approach
Part 2
Purification
Section 1

5. PROCESS DESCRIPTION
Water, microorganisms, nutrients (glucose) and air are fed into a bioreactor where at 37°C a fermentation takes place, yielding an enzyme and impurities. Biomass is separated in a disk-stack centrifuge and the liquid is stored in tank. It is then processed in a diafilter where the remaining biomass is removed (with a small loss of product). It is stored again and then loaded onto a PBA chromatography column where the enzyme binds and eluted using a WFI/NaCl mixture.

6. PROCESS DESCRIPTION Part 1 Fermentation Section Fermentation Unit
Bioreactor
Feed
Water
Microorganism
Nutrients (glucose)
air
Condition
37 °C
Output (yield)
Enzyme + Impurities
separated in a disk-stack
centrifuge and the liquid is stored in tank. (storage 1)
Biomass

7. Part 2 Purification Section1 Part 3 Purification Section 2
Diafiltration
Remove remaining Biomass
Blending/ Storage
Store product
(small loss of product)
Part 3
Purification Section 2
PBA Chromatography Column
Bind the enzyme
And eluted by using WFI/NaCl mixture
Blending/ Storage
Store product

8. Part 1 Fermentation Section
Mode of operation: batch processing
Component registration:
Glucose
Biomass
CO2
WFI (water for injection)
Enzyme (new-water as reference comp.)
Impurities ( new-water as reference comp.)

9. Process Flowsheeting for Fermentation Section
UNIT PROCEDURE
EQUIPMENT
DESCRIPTION
Fermentation
Vessel Procedure/ In a Fermentor
transformation of raw material into enzyme & impurities
Centrifugation
Centrifugation/ Disk-Stack
separation of biomass
Storage 1
Storage/ Bulk/ Batch/ in a Blending Tank)
Rename as: Storage 1 (Right click equipment/ Edit labels)
temporary product storage

10. PRODUCT INITIALISATION FOR FERMENTATION SECTION
Centifugation
Storage 1

11. FERMENTATION Initialising CHARGE operation
(right click on unit procedure (Fermentation)then click add /remove operations..
Add Charge -1, Charge-2, Heat-1, Ferment-1, Transfer-Out-1.

12. Duration: calculated based on constant heating rate 0.5 °c/min
Continue…
CHARGE OPERATION
DATA
CHARGE-1
Charge L of 100 L/min
CHARGE-2
Charge 1000kg 40kg/min
HEAT-1
Final temp :37 °C
Efficiency: 90%
Duration: calculated based on constant heating rate 0.5 °c/min
TRANSFER-OUT-1
duration same as Centrifugation
(use Master-Slave relationship)

13. Ferment-1 (Stoichiometric) Operation condition: Final temp: 37 °C
Continue…
CHARGE OPERATION
DATA
Ferment-1 (Stoichiometric)
Operation condition:
Final temp: 37 °C
Heat transfer agent: Cooling water
Process time: 36 hrs
Fermentor aeration: select air from stock mixture
(auto adjust)
Reaction (mass stoichiometry);
100Glucose + 80 O Water + 2 Enzyme +
3 Impurities + 80 CO2 +
40 Biomass
Reaction extent: 98% based on limiting component
Enthalpy: kcal/kg; ref. comp.: O2; ref. temp:37 °C
Emission: 100% for CO2 (select “Perform emission calculation” & “Set By User” for CO2)
Note: Leave other values as DEFAULT

14. FERMENT-1 Final temp: 37 °C Process time: 36 hr Reaction extent
Enthalpy data
Aeration setting:
Auto adjust for air
(stock mixture)
Mass stoichiometry

15. CENTRIFUGATION UNIT PROCEDURE OPERATING CONDITION MATERIAL BALANCE
UTILITIES
SCHEDULING
CENTRIFUGE-1 (default)
Equipment design based on: Solid Removal
Duration: 3 hr (Centrifugation time)
Component removal %: set by User
Solid component removal %: 98% for biomass; 0% for others
Solids Concentration in Solid Streams: 500 g/L
Exist temp:
15 °C (Set by User)
Agent:
chilled water
Start when Transfer-out of Fermentation
(P-1) starts

16. STORAGE 1 TRANSFER-IN-1: Operating conditions:
CHARGE OPERATION
DATA
TRANSFER-IN-1:
Operating conditions:
Transfer in using: outlet stream from centrifuge
Duration: same as Centrifuge (set by Master-Slave Relationship)
Scheduling: Start when Centrifugation (P-2) starts operation
STORAGE 1
Duration: to be determined
Scheduling: start when TRANSFER-IN-1 starts

17. the error message given
Let’s simulate the
flowsheet & solve
the error message given
(scheduling problem)

18. PURIFICATION SECTION 1 Please delete “STORE-1”
Right click on storage; then click on add/remove operations/delete storage/add TRANSFER-OUT-1
Please delete “STORE-1”
operation in P-3 & replace
it with a “Transfer-Out-1”

19. Process Flowsheeting for Purification Section 1
UNIT PROCEDURE
EQUIPMENT
DESCRIPTION
Diafiltration (DF)
Filtration/ Diafiltration
Removal of all leftover biomass from Storage 1
Remark: Storage (P-3) outlet needs to be deleted before new stream can be connected to the diafilter
Product Storage 2
Storage/ Bulk/ Batch/ In a Blending Tank
Rename as: Storage 2
Temporary product stage
(Note; Right click on equipment & select “Flip (reverse direction)” to turn the equipment into reverse direction

20. PROCESS DESCRIPTION ; Diafiltration
In diafiltration, water or some other solvent or buffer is added to the retentate to facilitate the removal of membrane-permeating species along with the water (or other solvent) during filtration.
The addition of water (or any other solvent) can be conducted either in batch or continuous mode.
recycle
Feed tank
In batch operation, permeable solutes are:
Cleared from the retentate by volume reduction (batch concentration);
Followed by re-dilution with water ( or other solvent); and
Re-concentration in repetitive steps

21. Diafiltration in SuperPro
In the current version of SuperPro Designer, batch concentration can precede and follow a continuous operation (true diafiltration)
Any number of batch concentration stages can be specified for each discontinuous operation.
In general, if the initial solution is dilute, a concentration step (to reduce the volume of the material) usually precedes a continuous diafiltration step.
Feed
tank
Recycle Loop
Permeate
(Filtrate)
Retentate
(Concentrate)
If the initial solution concentration is rather high, one usually
goes directly to continuous diafiltration

22. PRODUCT INITIALISATION FOR PURIFICATION SECTION-1
Diafiltration
Storage 2

23. DIAFILTRATION UNIT PROCEDURE OPERATING CONDITION UTILITIES SCHEDULING
DIAFILTER-1
(diafiltration)
Rejection coefficient (RC): biomass 100%, impurities 20%, enzyme 5%
Max. solid concentration in retentate: 600 g/L
Product denaturation (denaturation is due to shear forces during membrane filtration, common in bioprocessing):
Denaturation: 4%
Active product: enzyme
Denaturated product: impurities
Duration: 4 hrs (filtration time)
Diluant: water (auto adjust)
Diafiltration data: 5 (Volume Permeated)
Concentration data:
Prefiltration: YES, # of conc. stages: 1, Conc. Factor 5
Postfiltration: NO
Select “set by User”
Exit temperature = 15°C
Agent: Glycol
Specific power: 0.2 kW/m2
Start with TRANSFER-OUT of Storage 1

24. Storage1 (P-3) to follow the duration of Filtration inDiafilter
Continue…..
Additional task:
Set TRANSFER-OUT-1 of
Storage1 (P-3) to follow the
duration of Filtration inDiafilter
(P-4) using Master-Slave
relationship

25. STORAGE 2 TRANSFER-IN-1 Transfer in using: outlet stream from DF (P-4)
UNIT PROCEDURE
OPERATING CONDITION
SCHEDULING
TRANSFER-IN-1
Transfer in using: outlet stream from DF (P-4)
Start (scheduling) and duration (Operating condition: Master-Slave) same as DF
STORAGE 2
Duration: to be determined
Scheduling: start when TRANSFER-IN-1 starts

26. Simulate the Flowsheet
&
Solve The
Scheduling Error

27. PURIFICATION SECTION 2 Again, replace “STORE -1” operation in P-5 with
“TRANSFER-OUT-1”

28. Process Flowsheeting for Purification Section 2
UNIT PROCEDURE
EQUIPMENT
DESCRIPTION
PBA Chromatography
Equipment: Chromatography/ Adsorption/ PBA Chromatography
Description: binds and is elutes the enzyme using a WFI/ NaCl mixture (new mixture to be registered
Product Storage 3
Storage/ Batch/ In a Blending Tank
Rename as: Storage 3
temporary product storage
(Note: Right click on equipment & select “Flip (reverse
direction)” to turn the equipment into reverse direction

29. NEW MIXTURE REGISTRATION
We need a mixture of “NaCl/WFI(2M)” for this section, but this mixture is not found in the component database of SuperPro (verify this from Stock Mixture database)
2 ways of registering this mixture:
A) MODIFY FROM EXISTING MIXTURE
‘Register as NaCl (2M) & replace the water compound in this mixture with WFI’
B) REGISTER FROM SCRATCH
Register it from scratch & fill in the physical properties that you have

30. A)MODIFY FROM EXISTING MIXTURE
Path: Task/Edit Stock Mixtures
Make sure the
mass % is make
up into 100%
Highlight the water
component, delete &
replace it with WFI

31. B)REGISTER FROM SCRATCH
Path: Task/Edit stock Mixtures
Create new mixture
Choose this option if
you know the density
of the mixture
Choose this option to modify from an existing mixture (e.g. NaCl mixture)

32. Let’s try it …
(Always remember to save your work …)

33. PRODUCT INITIALISATION FOR PURIFICATION SECTION 2
PBA Chromatography
Storage 3

34. GENERAL DESCRIPTION : PBA CHROMATOGRAPHY
4 different PBA Chromatography
Column;
1)Column Loading (Load)
2)Column Washing (Wash)
3)Column Elution (Elute)
4)Column Regeneration (Regenerate)
Regenerate

35. COLUMN
DESCRIPTION
PBA column loading (Load)
Estimate the time for loading a column, track recovery yield, & estimate the number and size of columns required
Column washing (Wash)
Wash away the undesired impurities that trap in the column
Column elution (Elute)
A column may be used to bind either:
Product component(s); or
Impurity components
For retention of product components, for a component that binds to the resin, its amount in the product stream = (amount in the feed stream) x (binding fraction) x (elution yield)
All component present in the feed stream, that do not bind to resin, exit into the waste stream
Colum regeneration (Regenerate)
Regenerate the resin using a solution

36. CHARGE OPERATION
OPERATING
CONDITION
SCHEDULING
ADDITIONAL
TASK
LOAD-1
Loading flowrate:
200 cm/h (linear velocity)
Resin’s primary function: Retain Product (default)
Comp binding & yield;
-enzyme 100%, 90%
-impurity: 20 %, 30%
Info;
Binding refers to all components that bind to the resin;
Yield represents the percentage of bound material that ends up in the product stream.
Starts when Transfer-Out of Storage 2
For Storage 2:
Make sure that the “Storage-1” operation in Storage 2 is replaced by “TRANSFER-OUT-1”
Set TRANSFER-OUT-1 of Storage 2 to have the same duration as LOAD-1 using Master-Slave Relationship

37. CHARGE OPERATION
OPERATING
CONDITION
ADDITIONAL
TASK
WASH-1
Volume per cycle: 2 BV (bed volumes)
Wash stream: “Wash” stream which contains WFI (auto adjust)
Additional task: Delete “Equilibrate” operation in P-6
ELUTE-1
Eluant Volume:
Total Volume: 8 bed vol.
Volume in Product Stream: 2 bed vol
Flow rate Options: 200 cm/h (linear velocity)
Elution Strategy: Gradient
Key comp data:
Name: NaCl
Initial concentration: 0 mol/L
Final concentration: 1 mol/L
Eluant A: NaCl/WFI (2M) in stock mixture
Eluant B: WFI (auto-adjust)
REGENERATE-1:
Linear velocity: 300 cm/h
Volume per Cycle: 2 BV
Wash stream: “Regen” stream with WFI (auto-adjust

39. STORAGE 3 CHARGE OPERATION DATA SCHEDULING TRANSFER-IN-1:
Transfer from: PBA chromatography; using outlet stream from PBA chromatography
Start (scheduling) and duration (Operating condition: Master-Slave) same as ELUTE-1 in PBA chromatography (Q: why not the last operation?)
TRANSFER-OUT-1:
Transfer to: none; using outlet stream from Storage 3
Start when TRANSFER-IN-1 completed

40. Check your simulation results
Check the input to your PBA chromatography
Since we specify comp binding & yield for:
Enzyme to be 100%, 90%
Impurity: 20%, 30%
The amount of enzyme in the product stream: ___kg
The amount of impurities in the product stream should be: ___kg
Please check this out & verify this from your
simulation results.

41. Biochemical Case Study REPORT
Prepare a detail report of the BIOCHEMICAL CASE STUDY and attached together
your simulation result (Gantt Chart)