PTT303 /2 PROCESS MODELLING AND

SIMULATIONSEM 1 (2013/2014)

Biochemical Case Study

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.

OUTLINE FOR BIOCHEMICAL CASE STUDY

A sequential modular approach to solve for a

moderate complex flowsheet Some common unit operations in

biochemical industries:

FermenterDisk-stack centrifugationDiafiltrationChromatography

Sequential modular approach

Part 1Fermentation Section

Part 2Purification Section 1

Part 3Purification Section 2

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.

PROCESS DESCRIPTION

PROCESS DESCRIPTION

separated in a disk-stack centrifuge and the liquid is stored in tank. (storage 1)

Part 1Fermentation

Section

Fermentation Unit

Bioreactor

Feed •Water•Microorganism•Nutrients (glucose)•air

Condition 37 °C

Output (yield)

Enzyme + Impurities

Biomass

Part 2Purification Section1

Part 3Purification Section 2

Diafiltration Remove remaining Biomass

Blending/ Storage

Store product(small loss of product)

PBA Chromatography Column

Bind the enzymeAnd eluted by using WFI/NaCl mixture

Blending/ Storage

Store product

Mode of operation: batch processing Component registration:

GlucoseBiomassCO2

WFI (water for injection)Enzyme (new-water as reference comp.)Impurities ( new-water as reference comp.)

Part 1Fermentation

Section

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

PRODUCT INITIALISATION FOR FERMENTATION SECTION

FermentationCentifugationStorage 1

FERMENTATIONInitialising 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.

CHARGE OPERATION

DATA

CHARGE-1 Charge 10000 L of water @ 100 L/min

CHARGE-2 Charge 1000kg glucose @ 40kg/min

HEAT-1 Final temp :37 °CEfficiency: 90%Duration: calculated based on constant heating rate 0.5 °c/min

TRANSFER-OUT-1 duration same as Centrifugation (use Master-Slave relationship)

Continue…

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 O2 55 Water + 2 Enzyme + 3 Impurities + 80 CO2 + 40 BiomassReaction extent: 98% based on limiting componentEnthalpy: -3700 kcal/kg; ref. comp.: O2; ref. temp:37 °CEmission: 100% for CO2 (select “Perform emission calculation” & “Set By User” for CO2)

Note: Leave other values as DEFAULT

FERMENT-1Final temp: 37 °C

Process time: 36 hr

Aeration setting:Auto adjust for air

(stock mixture)

Mass stoichiometry

Enthalpy data

Reaction extent

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

STORAGE 1

CHARGE OPERATION

DATA

TRANSFER-IN-1:

Operating conditions:Transfer in using: outlet stream from centrifugeDuration: same as Centrifuge (set by Master-Slave Relationship)

Scheduling: Start when Centrifugation (P-2) starts operation

STORAGE 1 Duration: to be determinedScheduling: start when TRANSFER-IN-1 starts

Let’s simulate the flowsheet & solve

the error message given (scheduling problem)

PURIFICATION SECTION 1

Please delete “STORE-1” operation in P-3 & replace it with a “Transfer-Out-1”

Right click on storage; then click on add/remove operations/delete storage/add TRANSFER-OUT-1

Process Flowsheeting for Purification Section 1UNIT

PROCEDUREEQUIPMENT 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

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.

PROCESS DESCRIPTION ; Diafiltration

In batch operation, permeable solutes are:Cleared from the retentate by volume

reduction (batch concentration);Followed by re-dilution with water ( or other

solvent); andRe-concentration in repetitive steps

Feed tank

recycle

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.

If the initial solution concentration is rather high, one usually goes directly to continuous diafiltration

Feed tank

Recycle Loop

Permeate(Filtrate)

Retentate (Concentrate)

PRODUCT INITIALISATION FOR PURIFICATION SECTION-1

Diafiltration

Storage 2

DIAFILTRATIONUNIT

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: enzymeDenaturated 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 5Postfiltration: NO

Select “set by User”

Exit temperature = 15°C

Agent: Glycol

Specific power: 0.2 kW/m2

Start with TRANSFER-OUT of Storage 1

Continue…..

Additional task: Set TRANSFER-OUT-1 ofStorage1 (P-3) to follow

theduration of Filtration

inDiafilter (P-4) using Master-Slave relationship

STORAGE 2

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

Simulate the Flowsheet &

Solve The Scheduling Error

PURIFICATION SECTION 2

Again, replace “STORE-1” operation in P-5 with “TRANSFER-OUT-1”

(Note: Right click on equipment & select “Flip (reverse direction)” to turn the equipment into reverse direction

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

NEW MIXTURE REGISTRATIONWe 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

A)MODIFY FROM EXISTING MIXTURE

Path: Task/Edit Stock Mixtures

Highlight the water component, delete & replace it with WFI

Make sure the mass % is make up into 100%

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)

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

PRODUCT INITIALISATION FOR PURIFICATION SECTION 2

PBA Chromatography

Storage 3

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

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); orImpurity 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

CHARGE OPERATION

OPERATING CONDITION

SCHEDULING

ADDITIONALTASK

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

CHARGE OPERATION

OPERATING CONDITION

ADDITIONALTASK

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

STORAGE 3CHARGE

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

Check your simulation results

Check the input to your PBA chromatographySince we specify comp binding & yield for:Enzyme to be 100%, 90%Impurity: 20%, 30%

The amount of enzyme in the product stream: ___kgThe amount of impurities in the product stream should be: ___kg

• Please check this out & verify this from your simulation results.

Biochemical Case Study REPORT

Prepare a detail report of the BIOCHEMICAL CASE STUDY and attached togetheryour simulation result (Gantt Chart)