INTRODUCTION

Acrylamide (C3H5NO) is one of the most important chemical commodities in

the industrial world today. It is in great demand as it forms the starting material
for the production of a large range of chemical derivatives.
Acrylamide is present in two forms, crystalline and aqueous, but more than
often it is found in its crystalline form. The crystals are colourless to white and
free flowing. Acrylamide is soluble in water, methanol, ethanol, dimethyl ether
and acetone. It is insoluble in benzene and heptane. In general it is
incompatible with acids, bases, oxidizing agents, iron and iron salts. The
melting point of the chemical compound is 840C 850C and its boiling point is
about 1250C. 50% aqueous form of acrylamide is often the preferred form for
applications in which water can be tolerated.
Although acrylamide is suspected to have carcinogenic effects on humans, it
has many applications in a varied number of industries. For example,
acrylamide is widely used in the production of polyacrylamides (polymers
formed from acrylamide subunits), which are majorly used as flocculants for
clarifying drinking water and treating municipal and industrial effluents.
Polyacrylamides are also used in the cosmetic industry and as gels (SDSPAGE) in biotechnology laboratories. Acrylamides are also used as flow
control agents to enhance oil production from wells in the oil industry. Some
other applications of acrylamide include the production of dyes and chemicals
like N-Methylacrylamide, co-polymers used in production of contact lenses,
permanent press fabrics, sizing paper and textiles, etc. They are also used as
binders and retention aids for pulp and paper production. In the construction
business, acrylamide is used as a soil stabilizer and grouting agent for
construction of tunnels, sewers, wells and reservoirs. Adhesives, herbicidal
treatment of lakes and food packaging are other areas where acrylamide is
used.

MARKET OVERVIEW OF ACRYLAMIDE

Acrylamide was first produced on a commercial scale by American Cyanide
CO., Ltd in 1954. Several improvements have been made to the production
process by companies like Mitsui Toatsu, American Cyanamid, Dow and
Mitsubishi Rayon Co, Ltd to eliminate certain steps which lead to large

amounts of ammonium sulphate and as such by this elimination, leads to higher

concentration of aqueous acrylamide.
Mitsubishi Rayon Co, Ltd MRC Japan (formerly called Nitto Chemical
Industry) started enzymatic production of acrylamide on a 4000 ton/annum
production capacity. This increased to 10,000 tons/annum for a ten-year period
and has risen to about 20,000 tons per year since 1998 with improvements in its
production process, which is now, accepted as low-cost, high quality and less
harmful to the environment. MRC has licensed its improved technologies to
other countries and has plans to do this in companies worldwide.
Worldwide demand for acrylamide was 200, 000 tons in 1995, doubled in 2000
to 400,000 tons and was expected to grow at a 3-5% rate yearly from 2000
through 2005. Production of acrylamide exceeded 400,000 tons worldwide in
1999. The world acrylamide market was worth approximately $2 billion in
2006. SNF polydene is the global leader in acrylamide production.
93% of the worlds acrylamide capacity is accounted for by the United States,
Western Europe, Japan and China.
The following chart shows consumption of acrylamide by end use:

Source: http://www.sriconsulting.com/CEH/Public/Reports/606.2000

The largest end use market globally is wastewater treatment. The fastest
growing end use market segment is enhanced oil recovery and 93% of the
worlds consumption for this market is accounted for by China with expected
growth at an average rate of 5% between 2006-2011. The United States is the
leading end user of acrylamide in mining and Japan in pulp and paper industry.

CHEMICAL ROUTE VIA BIO-ROUTE

Apart from the above-mentioned advantages, the investment in equipment is

much lower compared to the chemical route. The production cost is also lower.
The high selectivity and low reaction temperature reduces the amount and
range of by-products.

Advantages of resting cell system over immobilized cells system for the
production of Acrylamide
Using resting cells for acrylamide production via hollow fiber reactor system is
simple, easy and stable when compared to immobilized cells in the same
system. Immobilized cells have a major drawback in their usage as biocatalyst
due to the breakage of the immobilized cell beads, which introduces impurities
into the production process. Free cells have a high and pure product quality
yield in comparison to Immobilized cells. Free cells require low energy
consumption and are more efficient than the immobilized cells. Using free cells
the enzyme activity was higher than immobilized cells. Free cells are less prone
to impurities in comparison to the immobilized cells. Using resting cells makes
the process inexpensive and the reaction time is drastically reduced. The
biocatalyst released is utilized to the optimum extent.

PROCESS DESCRIPTION

The poly-sulfone hollow fiber bioreactor is used in the bioconversion of

acrylonitrile to acrylamide. In this system, membrane separation and
bioconversion are coupled. The PS hollow-fiber membrane was chosen and
applied in this membrane bioreactor system, due to its advantages of high flux,
high anti-pollution ability, cross flow filtration mode and easy cleanup.
The bioconversion was carried out using free cells of Brevibacterium sp as a
biocatalyst in the membrane bioreactor. Industrially, this process takes about
17 hours, with a 99.9% conversion rate of acrylonitrile to acrylamide.
Therefore, this membrane bioreactor has a favorable foundation for industrial
application due to its high efficiency and ability to continuously produce
acrylamide using free resting cells.
MATERIALS AND METHODS
Microorganism: The used strain is Brevibacterium sp. CH2 that has a high
nitrile hydratase activity and a high acrylonitrile concentration tolerance.
Nitrile hydratases (NHases) are mononuclear iron or non-corrinoid cobalt
enzymes that catalyse the hydration of diverse nitriles to their corresponding
amides
NHases have been efficiently used for the industrial production of acrylamide
from acrylonitrile and for removal of nitriles from wastewater. Photosensitive
NHases intrinsically possess nitric oxide (NO) bound to the iron centre and its
photo dissociation activates the enzyme.

Microorganism and Culture

The culture medium was composed of 20 g/l glucose, 5 g/l yeast extract, 7.5 g/l
carbamide, 1 g/l monosodium glutamate, 0.5 g/l KH2PO4, 0.5 g/l K2HPO4,
0.5 g/l MgSO47H2O, 60 ppm enzyme activity inducer, pH = 7.5.
Large-scale fermentation was carried out in a 2m3 fermenter at 28oC 500 rpm
agitation for 96 hours. Aeration was achieved by bubbling filtered air into the
vessel. The cells harvested were used directly for determination of enzyme
activity or used as biocatalysts for bioconversion.
A steel reactor (6000 l) connecting a set of hollow-fiber membrane was the
main part of the membrane bioreactor system, and the free resting cells and
reactants were recycled in the coupling system by a positive displacement

pump. The enzymatic bioreaction, which bioconversed acrylonitrile to

acrylamide by NHase, was completed mainly in the glass reactor and then the
acrylamide solution was separated in the hollow-fiber membrane. In order to
maintain a constant system volume and ensure the stability of the product
concentration, pure acrylonitrile and water were proportionally fed into the
system according to the reaction rate. A cooling fluid in the cooling pipe
controlled temperature. Salt water was used as the cooling fluid to remove the
reaction heat in the system. The filtration rate of the membrane was adjusted in
the range 015 ml/min. Because there are large quantities of organic and
inorganic substrates contained in fermentation broth in addition to cells, such
as protein, nucleic acid, and glucide, the PS hollow-fiber membranes has the
ability to resist these impurities. Original experiments showed that the PS
hollow-fiber membrane could successfully remove 96.5% of protein in the
experimental system. It indicates that PS membranes with the 50 000 Da
(nominal molecule weight cut-off) are ideal for protein filtration and will
satisfy the purification of acrylamide to some extent.
(The protein was analyzed by the Bradford method.)
In the 17 h industrial scale experiment, the efficiency of enzyme activity and
acrylamide productivity was 10.96 g/cell/h and 187.2 g/cell, respectively,
which were almost double those of fed-batch immobilized cell modes.
The feasibility and superiority of this new membrane bioreaction process used
in the bioconversion of acrylonitrile into acrylamide was confirmed both in labscale and industrial-scale tests. The productivity and enzyme utilization of this
new process were increased by almost 100%, compared with those of the fedbatch and immobilized cell modes, which was the present bioconversion
system used in industrial processes. Therefore, this new membrane bioreactor
system has a favorable foundation for an industrial application because of its
high efficiency and continuous production of acrylamide using free resting
cells.

DOWNSTREAM SEPERATION OF AQUEOUS ACRYLAMIDE

The acrylamide produced in the reactor was first filtered using an ultra
filtration membrane. This membrane removes not only the dead cells and other
proteins, but also bacteria and other such contaminants. The purity level of
obtained by this process is high. The acrylamide is obtained in an aqueous form
that is then subjected to further downstream processes mentioned below:

Vacuum stripping-steam is injected into the tank .At high temperature (>25 to
50 C) all the volatile impurities in the product are vaporized and collected
outside in a condenser tank. The product remaining in the tank is partially
crystallized
Crystallizer-the partially crystallized product is left is cooled and pumped into
the crystallizer .the product is left in the crystallizer for a small period time, so
that the crystallization of the product is completed.
Centrifugecentrifuge the crystallized product at 4 degrees C and continuously
rinsed with wash water to remove any un crystallized acrylamide and nonvolatile impurities This wash water released from the centrifuge tank is
recycled back to the vacuum stripping step. This is done in order to crystallize
any un-crystallized acrylamide. A 10% purge is also considered to prevent
impurity accumulation
After centrifugation, 80% concentration of acrylamide is obtained which is
then dried using a spray dryer. We are using this dryer as it has the advantage
of counter flow heating which is the most effective form of drying. The
acrylamide is then packaged and sold in its crystallized form.

PROCESS FLOW DIAGRAM

http://www.fuda998.com/uploadimage/2009822163837680.jpg

www.capconindia.com/image

MASS BALANCE

In the mass balance, taking 100kg of feed per hour as the basis, 5kg of nutrients
and 95 kg of water is out into the fermenter. Assuming that 90%of the substrate
reacts, 0.2kg of un-reacted substrate comes out of the fermenter, along with the
biomass and 95kg of water. This mixture is then pumped into a centrifuge to
separate the cells from the other impurities. The cells will not be entirely free
of water. (This is because there will always be some place left between the
cells) and thus we get a concentrated broth with 71% cells and 29% water. The
stream that contains the wastewater and unused substrate is recycled into the
fermenter. A purge of ten percent is done to prevent the accumulation of the
impurities.

The concentrated broth, along with acrylonitrile and water (in a ratio which
was calculated from the literature) are then added to the reactor. In the reactor
98% of acrylonitrile is converted to acrylamide. We have assumed the figure
to be 98% as the process is being done at an industrial scale. After the
conversion the broth is passed through the ultra filtration unit to remove any
solid impurities along with any bacterial contamination and other proteins.
Dead cells and other cells are also removed from the filter and the stream is
recycled to recover any unreacted acrylonitrile. A 10% purge is done to prevent
accumulation of any impurities.

The acrylamide and water mixture obtained from the filter is then subjected to
downstream purification steps. The first step is vacuum stripping, where most
of the water from the mixture is lost as vapor. All of the volatile impurities are
removed in this step. Some of the acrylamide is crystallized, but the rest of the
crystallization is done in the crystallizer. The vacuum-stripping step
concentrates the acrylamide to about 80%. This concentrated mixture is then
pumped into the centrifuge to remove any non-volatile impurities like
trieneamides e.t.c. The concentrated acrylamide paste obtained after this step
has about 71.2% acrylamide and 28.8% water (assuming that not all the water
is vaporized)

The paste is then passed through a dryer to remove the water (28.8%).
Assuming that no drying process is 100% efficient, the dryer concentrates the
paste to about 98%. The water is lost as vapor. Thus the final amount of
acrylamide obtained is 19.55kg.

ENERGY BALANCE

The energy balance is based on the temperature differences in the different unit
operations and takes into account, the different specific heats of all the
components going into each unit.

COSTING OF EQUIPMENT (Bioreactor)

For costing of equipment and sizing we have assumed that we are using 5000L
Hollow Fiber Membrane Bioreactor and the cost is estimated as 1.2m dollars
which includes the cost of hollow fiber membrane. We are assuming that 35
batches will be produced per year to meet out the 60000 tones requirement; and
so the operating cost is estimated to be $6, 75,000 per year.

SAFETY MEASURES

1. Protective clothing should be worn by workers. As acrylamide is a

carcinogenic agent, human exposure to acrylamide should be prevented as
occupational hazards occur from dermal contact with solid monomer and
inhalation of dust and vapors.
2. Disposal: whatever cannot be saved for recovery or recycling should be
handled as hazardous waste and sent to a RCRA (Resource Conservation and
Recovery Act) approved incinerator or disposed in a RCRA approved waste
facility.
3. Acrylamide should be kept in a tightly closed container and stored in a cool
dry place and kept away from combustible and or reactive materials.

RECOMMENDATIONS
1. We recommend the use of both resting cells and immobilized enzymes
in the system; will make the production process more efficient. Since
immobilized cells cause a lot of problems (cleaning, lower activity) in
the process, it would be better to use immobilized enzymes.
2. We also recommend using counter flow mechanism in the filtration
unit as it would prove to be much more efficient at an industrial scale.

CONCLUSION
To conclude, a new strategy of membrane separation coupled with a bioconversion
process in which the free cells were used as a biocatalyst was found to be more
effective and more economical than the conventional chemical method of producing
acrylamide. Although immobilized cells are more advantageous than free cells in
other production processes, in this process it has been proved that free cells have more
advantages and thus can be used in the place of immobilized cells. The process is
found to have more conversion rate and the byproduct formation (acrylic acid) has
been much lower or no formation has been observed. Therefore, this new membrane
bioreactor system has a favorable foundation for an industrial application because of
its continuous production and high efficiency.