2

Uses of Citric Acid

Uses in the cosmetic industry
It is used in astringent lotions such as aftershave lotions because of its low
pH.
Citric acid is used in hair rinses and hair and wig setting fluids.
Miscellaneous uses in industry
In neutral or low pH conditions the acid has a strong tendency to form
complexes hence it is widely used in electroplating, leather tanning, and in
the removal of iron clogging the pores of the sand face in old oil wells.
Citric acid has recently formed the basis of manufacture of detergents in
place of phosphates, because the presence of the latter in effluents gives
rise to eutrophication.
Fermentation for Citric Acid Production
For a long time the production of citric acid has been based on the use of molasses
and various strains of Aspergillus niger and occasionally Asp. wenti.
Production by Penicillium is available, in practice are not used because of low
productivity.
Recently yeasts, especially Candida spp. (including Candida quillermondi) have
been used to produce the acid from sugar.
Japanese workers described a method to produce the acid by paraffins by bacteria
and yeasts. Among the bacteria were Arthrobacter paraffineus and corynebacteria;
the yeasts include Candida lipolytica and Candida oleiphila.
Fermentation with molasses and other sugar sources can be either surface or
submerged. Fermentation with paraffins however is submerged.
(a) Surface fermentation: Surface fermentation using Aspergillus niger may be done
on rice bran as is the case in Japan, or in liquid solution in flat aluminium or
stainless steel pans.
Special strains of Asp. niger which can produce citric acid despite the high content
of trace metals in rice bran are used. The citric acid is extracted from the bran by
leaching and is then precipitated from the resulting solution as calcium citrate.


Fermentation for Citric Acid Production
(b) Submerged fermentation: As in all other processes where citric acid is made
the fermentation the fermentor is made of acid-resistant materials such as
stainless steel.
The carbohydrate sources are molasses decationized by ion exchange, sucrose or
glucose. MgSO4, 7H
2
O and KH
2
PO
4
at about 1% and 0.05-2% respectively are
added (in submerged fermentation phosphate restriction is not necessary).
The pH is never allowed higher than 3.5.
Copper is used at up to 500 ppm as an antagonist of the enzyme aconitase which
requires iron.
1-5% of methanol, isopranol or ethanol when added to fermentations containing
unpurified materials increase the yield; the yields are reduced in media with
purified materials.
As high aeration is deleterious to citric acid production, mechanical agitation is
not necessary and air may be bubbled through. Anti-form is added.
The fungus occurs as a uniform dispersal of pellets in the medium.
The fermentation lasts for five to fourteen days.
Extraction
The broth is filtered until clear.
Calcium citrate is precipitated by the addition of magnesium-
free Ca(OH)
2
.
Since magnesium is more soluble than calcium, some acid
may be lost in the solution as magnesium citrate if
magnesium is added.
Calcium citrate is filtered and the filter cake is treated with
sulfuric acid to precipitate the calcium.
The dilute solution containing citric acid is purified by
treatment with activated carbon and passing through iron
exchange beds.
The purified dilute acid is evaporated to yield crystals of citric
acid.
Further purification may be required to meet pharmaceutical
stipulations.
7
The Distillery Flow Diagram: From Corn to Ethanol Plus Co-products
1 bushel
of corn
(56 lbs)
Starch
(32 lbs)
Milling
Fermentable
sugar
(36 lbs)
Liquefaction
Saccharification
Ethanol
(17.6 lbs)
+
DDGS
(17 lbs)
+
Heat
(7450 BTU)
+
CO2
(18.4 lbs)

Fermentation
Recovery
Dryhouse
8
Fermentation Fundamentals
Key Ingredients
Starch (sugar source)
Enzymes (breakdown of starches)
Yeast (conversion of sugar to ethanol)

Key Process Components
Enzyme addition
Yeast propagation
Fermentation cycle
Cooling system
Energy CO OH H C O H C
2 5 5 6 12 6
2 2
9
Process Description
Milling
Cooking and mixing
Milled corn and cook water
a-Amylase to break down long polymer chains (starch), reduce solids
Solubilization of sugars
Release of bound sugars: starch is bound to protein and fiber which is released during
cooking
Gelatinization: absorb water so enzymes can react with molecules
Liquefaction
Using a-Amylase to break down starches to shorter chain molecules
Saccharification and Fermentation
Batch processs (~ 50-60 hours)
Exothermic
Glucoamylase is used to break down short chained molecules to glucose
Yeast is added to convert glucose to ethanol
10
Water Balance
Methanator
Slurry Tank
Process Water
Liquefaction
Backset
Beer Feed
Milling
11
Fermentation
Slurry Tank
Liquefaction
Backset
Beer Feed
Fermenter
Yeast Propagation
Enzymes
12
Distillation
Separate Ethanol from non-fermentable
components
Dehydration
Azeotropic Distillation
Molecular Sieves
Non-Fermentables
Centrifugation
Evaporation
Drying
Process Description
13
Distillation
Beer Feed
Evap. Steam
Steam Flow
Sieve Back
Pressure
Rectifier Reflux Sieve Feed
Vaporizer Temperature
14
Dryer, Evaporator and TO
Steam Flow
Centrifuge Feed
Whole Stillage
Thin Stillage
Syrup Tank
Wet Cake
Pad
Evaporators
Dryers
15
Control Strategy
Ethanol Plant-Wide Optimizer
Dryer Evap TOAPC
&
Distill/Sieve APC
Fermentation APC
Slurry/Waterbalance
APC
Mv setpoints
Plant and constraint
Measurements (CV)
Energy,
feedstock,
Product
Values
$ costs
Separation:

- The biomass is separated by filtration
- The liquid is transferred to recovery process:
- Separation of citric acid from the liquid: precipitation

calcium hydroxide is added to obtain calcium citrate
tetrahydrate wash the precipitate dissolve it with
dilute sulfuric acid, yield citric acid and calcium
sulfate precipitate bleach and crystallization
anhydrous or monohydrate citric acid.
Industrial Production of Citric Acid
Industrial Production of Ethanol
Application of Ethanol:

- Medical

- Fuel

- Industrial
Ethanol Production by Anaerobic Bioprocesses
Glycolysis
- Microorganism: S. cerevisae for hexose
Candida sp. for lactose or pentose
Genetically modified E. coli

- Ethanol production is growth-associated with S. cerevisae.

- Medium requirements for high production
- Carbon source: sugar cane, starch materials (e.g. corn, wheat), cellulosic
materials (?!). yield: 0.51 g ethanol/g glucose.
- N, P, minerals.
- Anaerobic
- 100g/L glucose are inhibitory for yeast.
- 5% (v/v) of ethanol are inhibitory for yeast.
- pH:4-6 for 30-35
o
C.

Industrial Production of Citric Acid
Bioreactor: batch, continuous or with cell recycle

95% conversion of sugars with a residence time of

40 h in batch reactor
21 h in continuous reactor without cell recycle
1.6 h in continuous reactor with cell cycle
By-products: glycerol, acetic acid, succinic acid.
Industrial Production of Citric Acid
Separation:

- Distillation to obtaining 95% (w/w) of ethanol-water
mixture, followed by
- Molecular sieves to removing water from the
mixture to get anhydrous ethanol.
Ethanol Production

http://www.cleantechblog.com/uploaded_images/production_process-718492.jpg
Purification of Citric Acid
A typical method used for purification of citric acid from a
fermentation broth involves two major purification techniques:
precipitation and filtration. The following schematic displays a
generic citric acid purification scheme. The scheme will be
discussed in detail in the next few slides.





ffff
Prec
React citric acid
with calcium
carbonate
Filter
precipitate
React precipitate
with sulfuric
acid
Filter
precipitate
Purified Citric
Acid
Purification of Citric Acid
The citric acid broth from the production fermenter is highly
contaminated by leftover biomass, salts, sucrose, and water. First,
the citric acid must be reacted with calcium carbonate to
neutralize the broth and form the insoluble precipitate calcium
citrate. Calcium citrate contains about 74% citric acid. The
stoichiometric equation is as follows:

CaCO
3
+ Citric Acid CO
2
+ Calcium Citrate













Kirk 15



Contaminated Citric Acid
CSTR
Calcium Citrate as
a precipitate plus
contaminants
Calcium Carbonate, CaCO
3
Purification of Citric Acid
The calcium citrate is then washed, heated, and filtered to remove
any number of the contaminants. Depending on the specific
design of the purification scheme, filters can be placed before the
first reaction with calcium carbonate, in series between the two
precipitation reactions or in any other combination that works.
Also, it is important to choose the best kind of filter for what is
being removed. For simplicity, the filters here will remove larger
contaminants first (sucrose and salts) and the smaller
contaminants later.







Kirk 15



Calcium Citrate as
a precipitate plus
contaminants
Filter
Ex. Plate filter, Rotary
presses, rack-and-
frame presses

Calcium Citrate,
biomass, water
Calcium Citrate,
biomass, water
Purification of Citric Acid
To crack the calcium citrate precipitate, sulfuric acid is needed.
The temperature of this reaction should stay below 60C. The
reaction will produce free citric acid and a new precipitate,
calcium sulfate, which will need to be removed later. The
stoichiometric coefficients for this reaction are all one.










Kirk, 15
CSTR
Calcium sulfate as a
precipitate, free citric
acid, biomass, water
Sulfuric
Acid, SO
4
Purification of Citric Acid
In this filter, the calcium sulfate is washed away from the citric
acid and the leftover biomass is removed. Again, the
contaminants that were present in the fermentation broth can be
removed by additional filtration means, such as microfiltration or
ultrafiltration.













Kirk 15



Filter
Ex. Plate filter, Rotary
presses, rack-and-
frame presses

Citric acid,
water
Calcium sulfate as a
precipitate, free citric
acid, biomass, water
Purification of Citric Acid
Further Purification
Citric acid can be produced in two forms monohydrate and
anhydrous. These forms may require additional purification steps
to reach the desired purity.

1. Monohydrate
contains one water molecule for every citric acid
molecule
Requires repeated crystallization until water content is approx. 7.5-
8.8%

2. Anhydrous
Processed to remove all water from end product
Prepare by dehydrating the monohydrate citric acid product at a
temperature above 36.6C
Kirk 16,17
KICgroup 1



Citric Acid Lime/Acid
Fermentation Precipitation
Acidification Crystallization
Lime
Calcium
Citrate
Sugars Residue
Citric
Acid
Sulfuric
Acid
Water
Gypsum
Purification Conclusion
Once the product has been brought to the desired purity, it would
be sent to packaging and distribution.

In summary, there are many different methods and types of
equipment that make up a purification scheme. Some of the
more common types or purification were discussed in the first
half of this tutorial. The second half of the tutorial dealt with a
specific citric acid purification scheme that utilizes precipitation
and filtration to recover free citric acid from a contaminated
fermentation broth.






Citric
Acid
Purification
INDUSTRIAL ALCOHOL PRODUCTION
Ethanol (ethyl alcohol, grain alcohol) is a clear,
colorless liquid with a characteristic, agreeable odor.

In dilute aqueous solution, it has a somewhat sweet
flavor, but in more concentrated solutions it has a
burning taste.

Ethanol, the word, alcohol derives from Arabic al-
kuhul, which denotes a fine powder of antimony used
as an eye makeup. Alcohol originally referred to any
fine powder, but medieval alchemists later applied the
term to the refined products of distillation, and this
led to the current usage.
Uses of Ethanol
(i) Use as a chemical feed stock: In the chemical industry, ethanol is an
intermediate in many chemical processes because of its great reactivity as shown
above. It is thus a very important chemical feed stock.
(ii) Solvent use: Ethanol is widely used in industry as a solvent for dyes, oils, waxes,
explosives, cosmetics etc.
(iii) General utility: Alcohol is used as a disinfectant in hospitals, for cleaning and
lighting in the home, and in the laboratory second only to water as a solvent.
(iv) Fuel: Ethanol is mixed with petrol or gasoline up to 10% and known as gasohol
and used in automobiles.

Denatured Alcohol
All over the world and even in ancient times, governments have
derived revenue from potable alcohol. For this reason when alcohol is
used in large quantities it is denatured or rendered unpleasant to
drink.
The base of denatured alcohol is usually 95% alcohol with 5% water;
for domestic burning or hospital use denatured alcohol is dispended
as methylated spirit, which contains a 10% solution of methanol,
pyridine and coloring material.
For industrial purpose methanol is used as the denaturant.
In the United States alcohol may be completely denatured (C.D.A.
completely denatured alcohol) when it cannot be used orally because
of a foul taste or four smelling additives.
It may be specially denatured (S.D.A. specially denatured alcohol)
when it can still be used for special purposes such as vinegar
manufacture without being suitable for consumption.
Ethanol is a clean-burning, high-
octane fuel that is produced from
renewable sources.

At its most basic, ethanol is grain
alcohol, produced from crops such
as corn.
A bushel of corn weighs 56 pounds and will
produce at least 2.8 gallons of ethanol , 17
pounds of distillers grain & 18 Pounds of
CO
2
In 2005, 97 ethanol plants in 21 states
produced a record 3.904 billion gallons of
ethanol
Ethanol can be made by a dry mill process or a
wet mill process.

Most of the ethanol in the U.S. is made using
the dry mill method. In the dry mill process,
the starch portion of the corn is fermented
into sugar then distilled into alcohol.
The major steps in the dry mill process are:

1. Milling. The feedstock passes through a hammer
mill which grinds it into a fine powder called meal.

2. Liquefaction. The meal is mixed with water and
alpha-amylase, then passed through cookers where
the starch is liquefied. Heat is applied at this stage to
enable liquefaction. Cookers with a high temperature
stage (120-150 degrees Celsius) and a lower
temperature holding period (95 degrees Celsius) are
used. High temperatures reduce bacteria levels in the
mash.
3. Saccharification. The mash from the cookers is
cooled and the secondary enzyme (gluco-amylase) is
added to convert the liquefied starch to fermentable
sugars (dextrose).

4. Fermentation. Yeast is added to the mash to
ferment the sugars to ethanol and carbon dioxide.
Using a continuous process, the fermenting mash is
allowed to flow through several fermenters until it is
fully fermented and leaves the final tank. In a batch
process, the mash stays in one fermenter for about 48
hours before the distillation process is started.
5. Distillation. The fermented mash, now called beer, contains about
10% alcohol plus all the non-fermentable solids from the corn and
yeast cells. The mash is pumped to the continuous flow, multi-
column distillation system where the alcohol is removed from the
solids and the water. The alcohol leaves the top of the final column
at about 96% strength, and the residue mash, called stillage, is
transferred from the base of the column to the co-product
processing area.

6. Dehydration. The alcohol from the top of the column passes
through a dehydration system where the remaining water will be
removed. Most ethanol plants use a molecular sieve to capture the
last bit of water in the ethanol. The alcohol product at this stage is
called anhydrous ethanol (pure, without water) and is approximately
200 proof.
Manufacture of Ethanol
Ethanol may be produced by either synthetic chemical method or by fermentation.
Fermentation was until about 1930 the main means of alcohol production.
In 1939, for example 75% of the ethanol produced in the US was by fermentation,
in 1968 over 90% was made by synthesis from ethylene.
Due to the increase in price of crude petroleum, the source of ethylene used for
alcohol production, attention has turned worldwide to the production of alcohol by
fermentation.
Fermentation alcohol has the potential to replace two important needs currently
satisfied by petroleum, namely the provision of fuel and that of feedstock in the
chemical industry.
The production of gasohol (gasoline alcohol blend) appears to have received
more attention than alcohol use as a feed stock.
Nevertheless, the latter will also surely assume more importance if petroleum price
continues to ride.

Manufacture of Ethanol
Governments the world over have set up programs designed to conserve
petroleum and to seek other energy sources.
One of the most widely publicized programs designed to utilize a new
source of energy is the Brazilian National Ethanol Program. Set-up in 1975,
the first phase of this program aims at extending gasoline by blending it
with ethanol to the extent of 20% by volume.
The United States government also introduced the gasoline programme
based on corn fermentation in 1980 following the embargo on grain sales
to the then Soviet Union.

Substrates
The substrate used will vary among countries.
In Brazil sugar cane, already widely grown in the country, is the major
source of fermentation alcohol, while it is planned to use cassava and
sweet sorghum.
In the United States enormous quantities of corn and other cereals
are grown and these are the obvious substrates.
Cassava grows in many tropical countries and since it is high yielding
it is an important source in tropical countries where sugar cane is not
grown.
It is recognized that two important conditions must be met before
fermentation alcohol can play a major role in the economy either as
gasohol or as a chemical feedstock.
First, the production of the crop to be used must be available to
produce the crop without extensive and excessive deforestation.
Secondly, the substrate should not compete with human food.
Fermentation
The sterilized fermentable sugars are pumped or allowed to flow by gravity into
fermentation tanks and yeast is inoculated or pitched in at a rate of 7-15 x 10
6

yeast cells/ml, usually collected from a previous process.
These broths are inoculated with up to 5% (v/v) of thick yeast broth.
Although yeast is re-used there is still a need for regular inocula.
In general the inocula are made of selected alcohol-tolerant yeast strains usually
Sacch. cerevisiae grown aerobically with agitation and in a molasses base.
Progressively larger volumes of culture may be developed before the desired
volume is attained.
When the nitrogen content of the medium is insufficient nitrogen is added usually
in the form of an ammonium salt.
As in all alcohol fermentations the heat released must be reduced by cooling and
temperatures are generally not permitted to exceed 35-37C.
The pH is usually in the range 4.5-4.7, when the buffering capacity of the medium
is high.
Higher pH values tend to lead to higher glycerol formation.
When the buffering capacity is lower, the initial pH is 5.5 but this usually falls to
about 3.5. During the fermentation contaminations can have
Distillation
After fermentation the fermented liquor or beer contains alcohol as
well as low boiling point volatile compounds such as acetaldeydes,
esters and the higher boiling, fusel oils.
The alcohol is obtained by several operations.
First, steam is passed through the beer which is said to be steam-
stripped.
The result is a dilute alcohol solution which still contains part of the
undesirable volatile compounds.
Secondly, the dilute alcohol solution is passed into the center of a
multi-plate aldehyde column in which the following fractions are
separated: esters and aldehydes, fusel oil, water, and an ethanol
solution containing about 25% ethanol.
Thirdly, the dilute alcohol solution is passed into a rectifying column
where a constant boiling mixture, an azeotrope, distils off at 95.6%
alcohol concentration.
Distillation
To obtain 200 proof alcohol, such as is used in gasohol blending, the
96.58% alcohol is obtained by azeotropic distillation.
The principle of this method is to add an organic solvent which will form
a ternary (three-membered) azeotrope with most of the water, but with
only a small proportion of the alcohol.
Benzene, carbon tetrachloride, chloroform, and cyclohezane may be
used, but in practice, benzene is used.
Azeotropes usually have lower boiling point than their individual
components and that of benzene-ethanol-water is 64.6C.
On condensation, it separates into two layers.
The upper layer, which has about 84% of the condensate, has the
following percentage composition: benzene 85%, ethanol 18%, water
1%.
The heavier, lower portion, constituting 16% of the condensate, has the
following composition: benzene 11%, ethanol 53%, and water 36%.
Distillation
In practice, the condensate is not allowed to
separate out, but the arrangement of plates within
the columns enable separation of the alcohol. Four
columns are usually used.
The first and second columns remove aldehydes
and fusel oils, respectively, while the last two
towers are for the concentration of the alcohol.
A flow diagram of conventional absolute alcohol
production from molasses is given in Fig. 20.4
Distillation
Ethanol Dry-Grind
Grind bushel of corn (56 lb.), add water, make mash,
cook to kill bacteria, expose starch
Add enzymes for flow & to convert starch to sugar
Introduce Yeasts in Batch Fermenters---- produce beer-
--- distill the ethanol
Products:
ethanol (2.75 gal.) requires heat to distill
DDGS- (18 lbs.) generally requires drying
CO2-- ( 18 lbs. ) food grade
150 bushel corn yields 413 gallons of ethanol per acre,
2700 pounds of DDGS

Corn
Corn Dry-Milling Process Overview
Alpha-amylase enzyme
CO2
Yeast and
Gluco-amylase enzyme
Whole Stillage
Thin Stillage
Coarse Solids
Feed Industry Co-products
(Source: Kelly Davis, Chippewa Valley Ethanol Company)
Corn Cleaning
Hammermill Mix Slurry Liquefaction
Cooker
Centrifuge Evaporator
Fermentation
Dist illat ion
Ethyl
Alcohol
Conditioned
Distillers
Solubles
Distillers
Dried
Grains with
Solubles
Distillers
Wet Grains
Rotary Dryer
7. Denaturing. Ethanol that will be used for fuel must
be denatured, or made unfit for human consumption,
with a small amount of gasoline (2-5%). This is done at
the ethanol plant.

8. Co-Products. There are two main co-products
created in the production of ethanol: distillers grain
and carbon dioxide. Distillers grain, used wet or dry, is
a highly nutritious livestock feed. Carbon dioxide is
given off in great quantities during fermentation and
many ethanol plants collect, compress, and sell it for
use in other industries.
Distillers grain can be fed to livestock wet or
dry. Dried distillers grain (DDG) is the most
common variety. Drying the distillers grain
increases its shelf life and improves its ability
to be transported over longer distances. If a
consistent nearby market can be secured,
ethanol producers can supply the feed as wet
distillers grain (WDG). The wet form is not as
easily transportable, but the cost of drying the
product is removed.
Dried distillers grain with solubles (DDGS) is the
form available to the feed industry. The liquid that is
separated from the mash during the distilling process
is partially dehydrated into a syrup, then added back
onto the dried distillers grain to create DDGS.

DDGS is a high quality feedstuff ration for dairy
cattle, beef cattle, swine, poultry, and aquaculture.
The feed is an economical partial replacement for
corn, soybean meal, and dicalcium phosphate in
livestock and poultry feeds. Historically, over 85% of
DDGS has been fed to dairy and beef cattle, and
DDGS continues to be an excellent, economical feed
ingredient for use in ruminant diets.
The personal care products industry is one of the largest
users of industrial ethanol, or ethyl alcohol.

Check the labels hairspray, mouthwash, aftershave,
cologne, and perfume all contain large amounts of
alcohol by volume. Ethanol is also used in many
deodorants, lotions, hand sanitizers, soaps, and
shampoos.
Recovery of Ethanol by
Distillation process
Ethanol was recovered by simple distillation
method.
Sugarcane Source of Green Energy
JUICE
BAGASSE
LEAVES & TOPS
SUGAR
CO-GENERATION
OF ELECTRICITY
Ethanol
Flowsheet of Sugar and Ethanol
Production
J
u
i
c
e

Ethanol from Molasses and Juice

MASH
JUICE FOR SUGAR
FACTORY
SUGAR
ETHANOL
FROM
SUGARS
SUGARCANE
MILLING
FERMENTATION
DISTILLATION
MOLASSES
JUICE FOR ETHANOL
ETHANOL
FROM
MOLASSES
ETHANOL
DISTILLERY
Sugars = Suc + Glu + Fru
Milling Diagram
Usina Santo Antonio S. A. - Fluxograma de Processo - Setor de moendas
gua de
embebio
Tanque Pulmo
Peneira
Rotativa
Trocador de calor
regenerativo
Tanque de
cido
Fosfrico
Caldo clarificado para
destilaria
Bagacilho para lodo
dos decantadores
Circuito acar
Circuito lcool
Eletro-Im
Vinhaa
(Entrada)
Vinhaa
(Sada)
Caldo clarificado do
decantador de lcool
Caldo do circuito lcool
para os trocadores de calor
Legenda
Caldo de cana
guas industriais
Bagao
Produtos qumicos
Bagao para as
caldeiras
Caldo do circuito acar
para a sulfitao
Cana
gua de lavagemde cana
Picador 01
Picador 02
Esteira de cana picada
Mesa 18 - cana inteira
Bagao da peneira
Desfribador
Rolo alimentador
Espalhador
FERMENTATION PROCESS
The medium is inoculated with a suspension
of conidia of Penicillium chrysogenum.
The medium is constantly aerated and
agitated, and the mould grows throughout as
pellets.
After about seven days, growth is complete,
the pH rises to 8.0 or above, and penicillin
production ceases

STAGES IN DOWNSTREAM PROCESSING
Removal of cells

The first step in product recovery is the
separation of whole cells and other insoluble
ingredients from the culture broth by
technique such as filtration and
centrifugation.



ISOLATION OF BENZYL PENICILLIN

The PH is adjusted to 2-2.5 with the help of phosphoric or sulphuric
acids.
In aqueous solution at low PH values there is a partition coefficient in
favor of certain organic solvents such as butyl acetate.
This step has to be carried out quickly for penicillin is very unstable at
low PH values.
Antibiotic is then extracted back into an aqueous buffer at a PH of 7.5,
the partition coefficient now being strongly in favor of the aqueous
phase. The resulting aqueous solution is again acidified & re-extracted
with an organic solvent.
These shifts between the water and solvent help in the purification of
penicillin.



The treatment of the crude penicillin extract varies according to the
objective, but involves the formation of an appropriate penicillin
salt.
The solvent extract recovered in the previous stage is carefully
extracted back with aqueous sodium hydroxide.
This is followed by charcoal treatment to eliminate pyrogens and by
sterilization.
Pure metal salts of penicillin can be safely sterilized by dry heat, if
desired. Thereafter, the aqueous solution of penicillin is subjected
to crystallization.


FURTHER PROCESSING
For parental use, the antibiotic is packed in sterile vials as
a powder or suspension.
For oral use, it is tabletted usually now with a film
coating.
Searching tests (ex: for purity, potency) are performed
on the appreciable number of random samples of the
finished product.
It must satisfy fully all the strict government standards
before being marketed
The main stages of Penicillin production
are:

PRODUCTION OF PENICILLIN V
Phenoxy methyl penicillin
Addition of different Acyl groups to the
medium.
Phenoxyacetic acid as precursor instead of
phenyl acetic acid.

When penicillin was first made at the end of
the second world war using the fungus
Penicilium notatum, the process made 1 mg
dm
-3
.
Today, using a different species (P.
chrysogenum) and a better extraction
procedures the yield is 50 g dm
-3
.
There is a constant search to improve the
yield.

Antibiotics can be selectively toxic by
targeting such features as the bacterial cell
wall, 70S ribosomes, and enzymes that are
specific to bacteria.

In this way the human eukaryotic cells are
unaffected.
For example:
penicillin, ampicillin, amoxycillin, methicillin
Inhibits enzymes involved in synthesis of
peptidoglycan for bacterial cell wall, causing
cell lysis.
Bacteriocidal
Narrow spectrum- little effect on Gram
negative cells.
Downstream processing is relatively easy
since penicillin is secreted into the medium
(to kill other cells), so there is no need to
break open the fungal cells.

However, the product needs to be very pure,
since it being used as a therapeutic medical
drug, so it is dissolved and then precipitated
as a potassium salt to separate it from other
substances in the medium.
The resulting penicillin (called penicillin G)
can be chemically and enzymatically
modified to make a variety of penicillins with
slightly different properties.

These semi-synthetic penicillins include
penicillin V, penicillin O, ampicillin and
amoxycillin.
1. What is the Carbon source?
2. What is the nitrogen source?
3. What is the energy source?
4. Is the fermentation aerobic or anaerobic?
5. What is the optimum temperature?
6. Is penicillin a primary or secondary metabolite?
7. What volume fermenter is used?
8. Why isn't a larger fermenter used?
9. When is penicillin produced?
10. How long can it be produced for?
11. What was the first fungus known to produce penicillin?
12. What species produces about 60mg/dm3 of penicillin?
13. How did scientists improve the yield still further?
14. What is the substrate?
15. Why is batch culture used?
16. What are the processes involved in down-stream processing?
a)
b)
c)
17. Why can't penicillin be taken orally?
18. Name the form of penicillin which can be taken orally.
19. How does Penicillin kill bacteria?
20. Why are Gram negative bacteria not killed by penicillin?
lactose
yeast
glucose
aerobic
25 - 27C
secondary
40 200 dm
3
To difficult to aerate
40 hours after main increase in fungal mass
140 hours (180 40 hours)
Penicillin notatum
Penicillin chrysogenum
Genetic modification
Corn steep liquor
Secondary metabolite
Filtration of liquid
Extraction from filtrate by counter current of butylacetate
Precipitation by potassium salts
Destroyed by stomach acid
Penicillin V, ampicillin
Stops production of cell wall
Different cell wall