Crystallization & Whole broth

processing
 Crystallization is important as an industrial process because
of the number of materials that are and can be marketed are
in the form of crystals.
 Crystallization may be carried out from a vapor, from a
melt, or from a solution.
 More than 80% of the substances used in pharmaceuticals,
fine chemicals, agrochemicals, food and cosmetics are
isolated or formulated in their solid form.
 Crystallization is in general the last chemical purification
step in the production of ingredients.
Crystallization is the (natural or artificial) process of
formation of solid crystals precipitating from a solution,
melt or more rarely deposited directly from a gas.
 Crystallization is also a chemical solid-liquid separation
technique, in which mass transfer of a solute from the liquid
solution to a pure solid crystalline phase occurs.
Its extensive use is based on the fact that this single
operation is both a separation and a purification process
whereby a solid crystalline product can be isolated with high
purity and with relatively low capital and operating costs.
Crystals
A crystal may be defined as a solid composed of atoms arranged in
an orderly, repetitive array.
Crystals are grown in many shapes, which are dependent upon
downstream processing or final product requirements
Crystal shapes can include cubic, tetragonal, orthorhombic,
hexagonal, monoclinic, triclinic, and trigonal.
In order for crystallization to take place a solution must be
"supersaturated".  
Supersaturation refers to a state in which the liquid (solvent)
contains more dissolved solids (solute) than can ordinarily be
accomodated at that temperature
The crystallization process consists of two major events, nucleation
and crystal growth.
Nucleation
Nucleation is the step where the solute molecules
dispersed in the solvent start to gather into clusters, on
the nanometer scale (elevating solute concentration in
a small region), that becomes stable under the current
operating conditions.
the clusters reach a critical size in order to become
stable nuclei. This is dictated by the operating
conditions (temperature, supersaturation, etc)
Total nucleation is the sum effect of two categories of
nucleation - primary and secondary.
Primary nucleation
Primary nucleation is the initial formation of a crystal
where there are no other crystals present or where, if
there are crystals present in the system, they do not
have any influence on the process.
This can occur in two conditions
1. homogeneous nucleation
2. heterogeneous nucleation
Secondary nucleation
Secondary nucleation is the formation of nuclei
attributable to the influence of the existing microscopic
crystals in the magma.
first type of known secondary crystallization is
attributable to fluid shear, the other due to collisions
between already existing crystals with either a solid
surface of the crystallizer or with other crystals
themselves
Crystal growth
Once the first small crystal, the nucleus, forms it
acts as a convergence point for molecules of solute
touching - or adjacent to - the crystal so that it
increases its own dimension in successive layers.
Growth rate is influenced by several physical
factors, such as surface tension of solution,
pressure, temperature, relative crystal velocity in
the solution
Artificial methods
For crystallization to occur from a solution it must be
supersaturated
This can be achieved by various methods,
1. solution cooling,
2. addition of a second solvent to reduce the solubility
of the solute
3. chemical reaction
4. solvent evaporation
Applications
There are two major groups of applications for the
artificial crystallization process:
1. crystal production and
2. purification.
Equipment for crystallization
Tank Crystallizers
Forced circulation crystallizer
Scraped surface crystallizers
Circulating-magma vacuum crystallizer
Oslo crystallizer
Tank Crystallizers
This is probably the oldest and most basic method of
crystallization
Hot, saturated solutions are allowed to cool in open
tanks.
After crystallization, the mother liquor is drained and
the crystals are collected. 
Controlling nucleation and the size of the crystals is
difficult.  
The crystallization is essentially just "allowed to
happen"
Scraped surface crystallizers
One type of scraped surface crystallizer is the
Swenson-Walker crystallizer,
it consists of an open trough 0.6 m wide with a
semicircular bottom having a cooling jacket outside.
A slow-speed spiral agitator rotates and suspends the
growing crystals on turning.
The blades pass close to the wall and break off any
deposits of crystals on the cooled wall.
The Forced Circulation ("FC") crystallizer
the most common type of crystallizer in the industry
The average FC crystallizer evaporates solvent, thus increasing
the supersaturation in the process liquor, and causing
crystallization to occur.
Most conventional FC units operate under vacuum, or at slight
super atmospheric pressure.
The FC consists of four basic components: the crystallizer vessel,
heat exchanger, the circulating pump and the vacuum equipment
Slurry from the crystallizer vessel is circulated, in plug-flow
fashion, through the heat exchanger, and returned to the
crystallizer vessel again, where its supersaturation is relieved by
deposition of material on the crystals present in the slurry.
Circulating-magma vacuum crystallizer
The magma or suspension of crystals is circulated out of the
main body through a circulating pipe
The magma flows though a heater, where its temperature is
raised.
The heated liquor then mixes with body slurry and boiling
occurs at the liquid surface.
This causes supersaturation in the swirling liquid near the
surface, which deposits in the swirling suspended crystals until
they leave again via the circulating pipe.
The vapors leave through the top. A steam-jet ejector provides
vacuum.
DTB crystallizer is an example of this type of crystalliser
DTB (Draft Tube and Baffle) crystallizer
 Oslo-Krystal Cooling crystallizers
A small quantity of warm concentrated
feed solution enters the crystallizer
vessel at point A, located directly above
the inlet to the circulation pipe B.
Saturated solution from the upper regions
of the vessel together with The small
amount of feed liquor, is circulated by
pump C through the tubes of heat
exchanger D, which is cooled rapidly by
a forced circulation of water or brine.
On cooling the solution becomes
supersaturated, but not sufficiently for
spontaneous nucleation to occur
(metastable).
The supersaturated solution flows down
pipe E and emerges from the outlet F,
directly into a mass of crystals growing
in the vessel.
Whole broth processing
The concept of recovering a metabolite directly from
an unfiltered fermentation broth is of considerable
interest because of its simplicity, the reduction in
process stages and the potential cost savings.
It may also be possible to remove the desired
fermentation product continuously from a broth during
fermentation so that inhibitory effects due to product
formation and product degradation can be minimized
throughout the production phase.
Methods
Ion exchange resins
Reciprocating-plate extraction column
Dialysis
Expanded-Bed Adsorption
Resin method.
Ion exchange resins
 Ion exchange resins are polymers that are capable of exchanging
particular ions within the polymer with ions in a solution that is passed
through them
 The resins are prepared as spherical beads 0.5 to 1.0 mm in diameter.
 These appear solid even under the microscope, but on a molecular
scale the structure is quite open.
 This means that a solution passed down a resin bed can flow through
the crosslinked polymer, bringing it into intimate contact with the
exchange sites.
Extraction of Stretomycin
A process for adsorption of streptomycin on to a series of cationic ion-
exchange resin columns was developed directly from the fermentation broth,
which had only been screened to remove large particles so that the columns
would not become blocked This procedure could only be used as a batch
process.

Extraction of Novobiocin
The harvested broth was first filtered through a vibrating screen to remove
large particles.
The broth was fed into a continuous series of well mixed resin columns fitted
with screens to retain the resin particles, plus the adsorbed novobiocin, but
allow the streptomycete filaments plus other small particulate matter to pass
through.
The first resin column was removed from the extraction line after a
predetermined time and eluted with methanolic ammonium chloride to
recover the novobiocin.
Reciprocating-plate extraction column
 The Karr Reciprocating Plate Column consists of a series of
perforated plates with large diameter holes and high free cross
sectional area mounted on a reciprocated central shaft
 The reciprocating motion imparts energy to the liquids,
creating droplets and thus providing interfacial area so that
mass transfer can take place
 Along with the perforated plates are periodic baffle plates that
suppress axial mixing.
 The entire series of perforated and baffle plates is assemble in
a single cartridge, which is suspended from the drive motor on
top
 There are no internal bearings or guide bushings, and all
maintenance is performed outside of the column
 Dialysis
Removal of soluble impurities from solution by the use
of semipermeable membrane is known as dialysis
Solutes present in a solution(broth) can pass through a
semipermeable membrane.
Cycloheximide was extracted using methylene
chloride. Methylene chloride was circulated in a
dialysis tubing loop which passed through a fermentor.
Cycloheximide was extracted into methylene chloride.
The product yield increased by almost double by this
dialysis-solvent extraction method.
Resin Method
Sterile beads of an acrylic resin, as dispersed beads
or beads wrapped in ultrafiltration method, were
put in fermentors 48 hours after inoculation.
Some of the cycloheximide formed in broth was
absorbed in resin.
Recovery of antibiotic from resin is achieved by
solvents or by changing temperature.
Electrodialysis(ED)
Electrodialysis(ED) is a well known separation process
where ionized compounds are separated from non
ionized compounds in aqueous solutions based on
transport through ion exchange membranes in an
electric field.
Since in a fermentation broth the lactate salt is ionized,
whereas the carbohydrates and proteins and amino
acids are either non ionized or weakly ionized,
recovery and purification of lactate salts from a
fermentation broth by electrodialysis is feasible.
Expanded-Bed Adsorption Theory

Expanded-Bed Adsorption Theory

When the resin has packed in the column, the beads are close together (1). As the
column is fluidized, the resin beads establish a concentration gradient (2). The sample
feedlot is injected, and particulates and cell debris (green dots) move past the resin and
out of the column, while the compound of interest (red dots) interacts with the beads
(3). The column is then repacked, the flow is reversed, and the compound is eluted
from the beads (4).