Downstream Process

Miranda Putera Ria

Revitasari Rakhmat Widiastuti
051510… 05161061 051610…

MEMBER OF GROUP
 OUTLINE
1 INTRODUCTION
2 SOLID-LIQUID SEPARATION :
FILTRATION, CENTRIFUGATION

3 CELL RUPTURE
INTRODUCTION
 WHY WE NEED TO
LEARN ABOUT THIS?
 After successful fermentation or enzyme
reactions, desired products
must be separated and purified. This final step is
commonly known
as downstream processing or bioseparation,
which can account for
up to 60 percent of the total production costs,
excluding the cost of
the purchased raw materials
 The fermentation products can be the cells themselves (biomass),
components within the fermentation broth (extracellular), or those
trapped in cells (intracellular), examples of which are listed in
Table 10.1.
 As shown in Figure 10.1, if the product of our interest is
the cell, cells are separated from the fermentation -broth and then
washed and dried. In the case of extracellular products, after the cells
are separated, products in the dilute aqueous medium need to be
recovered and purified.
 UNIQUE CHARACTERISTIC OF
BIOSEPARATION PRODUCTS
THE PRODUCTS ARE IN DILUTE CONCENTRATION
1 IN AN AQUEOUS MEDIUM.

THE PRODUCTS ARE USUALLY

2
TEMPERATURE SENSITIVE.
THERE IS A GREAT VARIETY OF
3 PRODUCTS TO BE SEPARATED.
 UNIQUE CHARACTERISTIC OF
BIOSEPARATION PRODUCTS
THE PRODUCTS ARE IN DILUTE CONCENTRATION
1 IN AN AQUEOUS MEDIUM.

THE PRODUCTS ARE USUALLY

2
TEMPERATURE SENSITIVE.
THERE IS A GREAT VARIETY OF
3 PRODUCTS TO BE SEPARATED.
 UNIQUE CHARACTERISTIC OF
BIOSEPARATION PRODUCTS
THE PRODUCTS CAN BE INTRACELLULAR, OFTEN
4 AS INSOLUBLE INCLUSION BODIES

THE PHYSICAL AND CHEMICAL PROPERTIES OF

5 PRODUCTS ARE SIMILAR TO CONTAMINANT

EXTREMELY HIGH PURITY AND HOMOGEINITY

6 MAY BE NEEDED FOR HUMAN HEALTH CARE
SOLID-LIQUID
SEPARATION
FILTRATION
 Filtration separates particles by
forcing the fluid through a filtering
medium on which solids are
deposited.
There are generally two major types of filters: pressure and vacuum
filters. The two types of filters most used for cell recovery are the
filter press and rotary drum filters.
 A filter press is often employed for the small-scale
separation of bacteria and fungi from broths. For large-scale
filtration, rotary drum filters are usually used. A common filter
medium is the cloth filter made of canvas, wool, synthetic fabrics,
metal, or glass fiber.
 Assuming the laminar flow across the filter, the rate of filtration
(dVf/dt) can be expressed as a function of pressure drop ~p by the
modified D'Arcy's equation
 the filtration rate is proportional to the pressure drop and
inversely proportional to the filtration resistance L/k, which is the
sum of the resistance by the filter medium Rm and that by the cake Rc
FILTRATION
CENTRIFUGATION
Centrifugation is an alternative method when
the filtration is ineffective, such as in the
case of small particles. Centrifugation
requires more expensive equipment than
filtration and typically cannot be scaled to
the same capacity as filtration equipment.
 Two basic types of large-scale centrifuges are the tubular and the
disk centrifuge. The tubular centrifuge consists of a hollow cylindrical
rotating element in a stationary casing. A typical tubular centrifuge has
a bowl of 2 to 5 in. in diameter and 9 to 30 in. in height The disk
centrifuge consists of a short, wide bowl 8 to 20 in. in diameter that
turns on a vertical axis
 The expression for the terminal velocity can be derived from the
balance of the forces acting on a particle, which is applicable when the
Reynolds number is less than 1
CENTRIFUGATION
CELL RUPTURE
 Once the cellular materials are separated, those with
intracellular proteins need to be ruptured to release their
products. Disruption of cellular materials is usually difficult
because of the strength of the cell walls and the high osmotic
pressure inside. The cell rupture techniques have to be very
powerful, but they must be mild enough so that desired
components are not damaged.
CELL RUPTURE
 CELL RUPTURE

1 PHYSICAL METHODS

2 CHEMICAL METHODS

3 BIOLOGICAL METHODS
 CELL RUPTURE

1 PHYSICAL METHODS

2 CHEMICAL METHODS

3 BIOLOGICAL METHODS
 PHYSICAL METHODS

1 MILLING

2 HOMOGENIZATION

3 ULTRASONICATION
 PHYSICAL METHODS

1 MILLING

2 HOMOGENIZATION

3 ULTRASONICATION
 CHEMICAL METHODS
1 SURFACTANTS
2 ALKALIS
3 ORGANIC SOLVENTS
4 OSMOTIC SHOCK
 CHEMICAL METHODS
1 SURFACTANTS
2 ALKALIS
3 ORGANIC SOLVENTS
4 OSMOTIC SHOCK
 BIOLOGICAL METHODS

1 ENZYMATIC DIGESTION
 BIOLOGICAL METHODS

1 ENZYMATIC DIGESTION
THANK YOU FOR YOUR
ATTENTION
ANY QUESTION