CELL
DISRUPTION
Oleh :
Yustina (NPM 1706123141)
Fitria Afriani (NPM 1706122901)
Mata Kuliah Bio Separasi
Outline
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Introduction
Let’s start with the first set of slides
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Introduction (cont’d)
Cell disruption is an essential part
of biotechnology and the
downstream processes related to
the manufacturing of biological
products
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Introduction (cont’d)
So, now let’s see the structure of a cell
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Plant Cell vs Animal Cell
So, now let’s see the structure of a cell
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Introduction (cont’d)
So, now let’s see the structure of a cell
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Method of Microbial Disruption
Different cell have different structures,
hence they require different methods
for disruption
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Ideal Technology Characterization
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Ideal Technology Characterization (cont’d)
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Methods of Microbial Disruption
Non Mechanical
Non Mechanical
Physical Methods
Thermolysis, Decompression,
Chemical Methods
Solvent, Detergents
Osmotic Shock
Mechanical Methods Non Mechanical
High Pressure
Homogenization, French
Biological Methods
Press, Sonication, Bead Mill, Enzymes,
Grinding
Cell
Disruption 12
Product of interest : Intracellular or Extracellular ?
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1. Mechanical Methods
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Disadvantages Cell Disruption Methods
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a. High Pressure Homogenization
Is a non-thermal technology that
has been widely studied as a
partial or total substitute
for thermal food processing.
HPH generally consist of a high
pressure pump, mostly in the
form of a one to three-piston
plunger pump which can be
electrically or pneumatically
actuated an interaction
assembly and a cooling unit.
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cont’d....
Advantages :
• Customizable forces
to optimize results
• Hot and Cold
homogenization option
• Smaller particle sizes
than other equipment
• Aplikasi pada industri
susu, pembuatan nano
emulsi, dll
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a. High Pressure Homogenization (cont’d)
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a. High Pressure
Let’s start with the first set of slides
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b. French Press
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cont’d...
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c. Ultra sound/sonication
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Let’s start with the first set of slides
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d. Bead Mills
This Method is affected by :
- Agitator speed
- Beads size
- Cell suspension concentration and flow rate
- Agitator disc design 24
cont’d....
a. High Pressure
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Thermolysis
Thermolysis has shown potential in becoming more
common in large scale production.
Periplasmic proteins in G(-) bacteria are released
when the cells are heated up to 50ºC.
Cytoplasmic proteins can be released from E.coli
within 10min at 90 ºC.
Improved protein release has been obtained after
short high temperature shocks, than when at longer
temperature exposures at lower values.
The results are highly unreliable, as the protein
solubility changes with temperature fluctuations.
Decompression
The cell suspension is mixed with
pressurized subcritical gas for a
specified time, depending on the cell
type.
The gas enters the cell and expends
on release, causing the cell to burst.
Decompression has been used in
small scale laboratories for the
disruption of E.coli.
Osmotic Shock
Exposure of cells to either high or low salt concentration causes cell disruption
Title???
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Title???
Let’s start with the first set of slides
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a. High Pressure
Let’s start with the first set of slides
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Detergents
Anionic :
sodium dodecyl
sulfate (SDS)
Non-ionic :
triton X100
Cationic : ethyl
trimethyl
ammonium
bromide
Cell disruption with detergents. Detergents interact with cell membrane
compounds which will lead to disassembly of cell membrane.
Solvents
Solvents which can
be used for cell Leads to
lysis include Ekstract cell release of
alchohols, wall’s lipid intracellular
dimethyil sulfoxida, components components
methyl ethyl ketone
or toluene
Enzymes
Enzymes degrade the cell wall components which will lead to release of
intracellular compounds.
Application
Cont’d.... Combined Enzymatic and Mechanical Cell Disruption and
Lipid Extraction of Green Alga Neochloris oleoabundans
Cont’d....
Cont’d....
Enzymatic lysis
of microbial cells
conclusion
Cell disruption is an essential part of biotechnology and the
downstream processes.
Metode cell disruption terdiri dari mechanical methods meliputi high
pressure homogenization, french press, sonication, bead mill, grinding,
non mechanical physical methods meliputi thermolysis,
decompression, osmotic shock, non mechanical chemical methods
meliputi solvents and detergents, non mechanical biological methods
meliputi enzymes.
Application of cell disruption is ...... Excretion of .....
