DOWNSTREAM (cell)
Introduction of Boprocess Technology
Nur Istianah
22/11/2018 Nur Istianah-PTB Downstream process- 2016
Flotation
• Flotation is another method to remove solid (cells) from fermentation broth, using air bubbles to float protein.
• We may use different kinds of high shear-force devices to make homogeneous solutions for liberating intracellular enzymes.
• Non-mechanical methods are also used to break down the cell wall and to release intracellular enzymes or proteins.
Aeration
• Particulates can be removed from aqueous suspension by attachment to rising air bubbles.
• This method is known as flotation, which is widely used for recovery of small particlesfrom aqueous suspension minerals.
• A related technique, for flotation, uses air–water
surface tension to strip out proteins from the broth
solution and accumulate them in a high protein.
Cell recovery
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Aeration
• Flotation is applied to concentrate Acinetobacter cerificans for production of SCP. Acinetobacter cerificans is a suitable microorganism used in the production of SCP.
• The synthesised protein is concentrated by aeration
using the flotation method
Cell recovery
• Downstream processing of fermentation broths usually begins with separation of cells by filtration or centrifugation.
• A combination of bioprocesses is required for product recovery. Separation of products such as ethanol, citric acid and antibiotics is extracellular but isolation of enzymes inside the cells requires cell rupture.
• Cells are broken down by lysis of the cell wall.
Cell disruption is used for downstream product recovery. Removal of biomass from the extracted product is necessary.
Cell disruption
• Disrupsi sel adalah proses rusaknya dinding sel
yang menyebabkan keluarnya senyawa
biokimia dari dalamnya. Perusakan dinding sel
ini biasanya disertai dengan lisis atau
robeknya dinding sel.
Methods
• Mekanis
• Metode ini disertari dengan penggunaan energy mekanik seperti penggerusan biomassa disertai abrasi, pengadukan dengan kecepatan tinggi, pemompaan dengan tekanan tinggi, dan ultrasonik.
• Non-mekanis
• Metode ini tidak disertari dengan penggunaan energy
mekanik melaikan dengan bantuan bahan kimia
ataupun energy non-mekanik. Beberapa metode non-
mekanik yang biasa digunakan yaitu terapi osmotik,
pembekuan, thawing, perusakan enzimatis, serta
menggunakan bantuan pelarut atau deterjen.
Role of cell disruption
• Disrupsi sangat sering digunakan untuk recovery produk dari biomassa termasuk isolasi enzim dari sel secara intraseluler.
Adapun biomassa yang keluar dari ekstraksi
atau recovery kemudian dipisahkan dan
dikeluarkan sebagai produk samping.
Mechanism
• Lysis of cells for intracellular product recovery
• Extraction of lysed or ground cells
• Removal of unconverted soluble substrate
• Removal of biomass from extracted
product
Mechanism
• The choice of filtration (batch,
continuous vacuum, cross flow, etc.)
• Centrifuge (vertical rotor, horizontal rotor)
• Sedimentation (and/or
coagulation)
Methods of cell lysis
• Osmotic shock • Freezing
• Solvent • Detergents
• homogeniser • organic solvent
extraction
22/11/2018 Nur Istianah-PTB Downstream process- 2016
CASE STUDY: PROCESS INTEGRATION OF CELL DISRUPTION AND FLUIDISED BED ADSORPTION FOR THE RECOVERY OF LABILE INTRACELLULAR ENZYMES
22/11/2018 Nur Istianah-PTB Downstream process-
2016
Case study
Case study
Case study
Cell recovery
• Application of charges and filtration may separate protein very efficiently.
• Electrokinetic deposition uses voltage gradients
of 1050 V/cm to produce solid biomass with
densities of up to 40% w/v.
Case study
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