Large Scale Simulation of Itaconic Acid Production from

Aspergillus terreus using SuperPro Designer

Laura Nieto1, Christian Rivera1 and German Gelves1*

1
University Francisco de Paula Santander. Av Gran Colombia 12E-96. Cucuta, Colombia
*Corresponding author e-mail:germanricardogz@ufps.edu.co

ABSTRACT
Itaconic acid is obtained from Aspergillus terreus fermentation. It is used in a wide number of sectors such as
industry, agriculture and health This substance is important to replace petroleum products However, its
production is expensive and this hinders the study in the acid production. Simulation is a great since it allows
researchers to make a more effective evaluation and self-assessment, reducing costs and avoiding losses. The
objective of this article is to perform a Itaconic acid production simulation on industrial scale and a
subsequent improvement to reduce costs. All this using SuperPro Designer software to demonstrate the
advantages of simulation. As initial conditions 90g / L of glycerol, 1g / L of NH 4NO3 and 1.25 of KH2PO4 is
taken as the media composition reported fro references. Separation and purification methods are performed to
obtain 500 mg units at the end of the process. it is possible to obtain a production yield of 2893.95 containers
of itaconic acid crystals per hour with an operating cost of 60,425,000USD per year and through an
improvement to the process these costs are reduced to 18,781,000USD maintaining the same production
performance. Also, the cost of manufacturing per container of 500 mg of itanic acid crystals is approximately
80 % cheaper those calculated without the improvement.

Keywords: Simulation, itaconic, Software, Glycerol, Costs

INTRODUCTION
Itatonic acid is an acid that in aqueous solution donates 2 H+ protons per molecule (diprosthetic acid), has
five carbons and presents 2 carboxyl groups. This is also known as methylensuccinic acid. It is used in a wide
range of sectors such as industry, agriculture and health (García,2015). This substance has certain
characteristics that have managed to generate a great interest in the world market, to polymerization, the
carboxyl group that owns the itanic acid confers this property. Itatonic acid has been studied, its methods of
obtaining, as well as its polymerization to synthesize materials that are easy to degrade and environmentally
friendly which is what is most needed today to correct the situation of contaminants that is attributed to
plastics. Its structural similarity to various acrylic derivatives has generated some interest in the polymer
industry, where it is used to produce mostly hydrogels, polyesters, plastics, artificial glass, paints, textiles and
glues. In addition, it is used in medicine, cosmetics, lubricants, plusers, herbicides and wool modifiers and in
the manufacture of detergents, water treatments, dispersants and adhesives. (Martinez, 2017). Currently
substances such as natural gas and Kurdish oil remain the commodity of many chemicals, the constant search
for renewable energy and that of the environment has prompted us to look for alternatives that replace these
wastes fossils base of many chemicals. It has been found that this acid can be obtained by fermentation of
sugars by Aspergillus terreus, thermotolerant fungus, with optimal growth in the range of 35-40oC, which
makes the process temperature close to room temperature, avoiding high energy expenditures. It is common
to find it in tropical and subtropical climates as a contaminant at food storage sites. (Rodriguez, 2018).
Itatonic acid is an important chemical platform that has a wide range of potential and real applications. It can
be used to replace a wide range of petroleum products that will reduce dependence on oil and harmful
environmental effects, yet it only occupies a niche market due to its high cost relative to acrylic acid and other
alternatives, which limits their use to low-volume markets. Today the largest producer of itanic acid is China,
exporting almost 78% of the world's itanic acid. Likewise, the South of America is the place where there are
the greatest imports of itanic acid, exports have been led by China, India, united states, Japan, South Africa
and united kingdom(García,2015). Global production is estimated to be 80000 tonnes per year at around price
and production is expected to grow by 5.5% each year between 2016 and 2023(Rodriguez, 2018). We also
find countries where there is not enough information or technology to produce industrial lycranic acid, and its
production is expensive and makes research difficult in this area. In this digital age new technologies and
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advances in computing have been developed, which are tasked with facilitating our work, one of these tools is
simulation, which allows us to obtain a variety of results and analyze multiple solutions for determine the
profitability of a process. Simulation is a great advantage because it allows researchers to make a more
effective assessment and self-assessment, identify gaps and propose possible optimizations in the processes,
the exercises can be performed again and again leading to the feedback and learning, saving time and
reducing costs (Petrides, 2014). This paper seeks to propose a design for obtaining industrial scale of itaconic
acid from Aspergillus terreus by simulation using SuperPro Designer software.

MATERIALS AND METHOD

Glucose and glycerol have been reported as the substrate with which you achieve better results. The
appropriate conditions for the development of the microorganism are; temperature (30-40oC), continuous
aeration, low starting pH (3-5), low operating pH( 2.2-3.8) high glucose concentrations(10-20%), enough
nitrogen, high concentration of high concentration of magnesium sulfate (0.5%), low phosphate content for
limit the growth of mycelium and adequate, trace levels of metals such as zinc, copper and iron. for an
industrial production of itanic acid, the following processes are generally proposed :

Figure-1. Unit operation for Itaconic Acid Production.

Isolation
For the isolation of Aspergillus terreus has been reported in the literature found in tropical and subtropical
climates, can be isolated from wood, leather, decaying plant material and food products such as stored grains,
potatoes, cereals, fruits and nuts. (Rodriguez, 2018)

Inoculum Preparation
In a process performed by M.Juy, J, Ears and M.lucca the microorganism was cultured in agar malt extract at
a temperature of 3o BC until sporulation was obtained. Petri boxes with agar were washed with 10 ml of
sterile saline and Tween. Then the Erlenmeyer containing 100mL of each medium that was prepared (7 in
total) were inoculated with an aliquot of the suspension of concentrated spores in order to obtain 5x107
spores/mL of production medium. Erlenmeyers stirred at 200rpm at 30ºC for 24hours in an orbital shaker.
The resulting pellets were used as inocules for fermentation.
Table-1. Composition of the media used (Juy, et al, 2010).

Table 2. Results obtained from each medium (Juy, et al, 2010).

In the previous process it was obtained that the lowest concentration of acid obtained was using the M2a
medium, the composition of said medium will be taken as a reference to start the process and it is intended to
reach a final concentration and then proceed to a purification and filling process.

Inoculation
Based on the work illustrated above, we proceed to continue with the possible industrial escalation, for this
will be made use of an inoculation train using 4 fermenters. The initial inoculation tank will be fed with the
M2a medium, with compositions of 90g/L glycerol, 1g/L of NH4NO3 and 1.25 of KH2PO4.

Fermentation
Continuous fermentation will take place in which an open system is established. The sterile nutrient solution
is continuously added to the bioreactor and an equivalent amount of used nutrient solution, with
microorganisms, is removed simultaneously from the system. It will be carried out continuously in order to
recirculate the medium to take advantage of the medium that was not used by the microorganisms of the first
diet. The yield of biomass from substrate and product from substrate to be considered for this process are Yx:s
0.27 g biomass/g substrate Yp:s 0.44 g IA/g substrate(Juy, et al, 2010). The overall reaction equation in each
fermenter will be:

Glycerol+ O2 → Biomass+ Itaconic Acid +CO2 + H 2 O

(1)
Separation
For separation a series of processes are proposed such as evaporation, filtration, crystallization. After
inoculation train it was proposed to use a double-acting evaporator to allow the separation and extraction of
water from Itacnico acid and to increase the concentration of acid. Subsequently it is recommended to use a
liquid liquid extractor using benzene as solvent for the separation of Itaconic acid from other components in
liquid solution, subsequently for removal of other solid compounds, a rotary filter is used which allows for
greater effectiveness in separating and retaining particles of solity, after this the filtered liquid is subjected
again to a double-acting evaporator this time to remove the residual benzene from the previous liquid-liquid
extraction, in this way it is subjected can concentrate the itaconic acidto optimal conditions for purification.

Purification
A crystallizer is used, this unit operation allows to obtain in solid state the dissolved substance in the form of
crystals, allowing to remove the waste water from the process and obtain the extracellular metabolite as pure
as possible. The advantage of this operation is that it also allows to obtain the product in a packageable
presentation for commercialization.

RESULTS AND DISCUSSIONS

The goal for this work is to perform a Itaconic acid production simulation on industrial scale and a
subsequent improvement to this to reduce costs, all this using SuperPro Designer software to demonstrate the
advantages of simulation. As initial conditions 90g / L of glycerol, 1g / L of NH 4NO3 and 1.25 of KH2PO4 will
be taken as the composition of the medium, reported in works carried out by other authors. Figure 2 shows the
unit operations for large scale simulation of itaconic acid usinf Super Pro Designer Software.

Figure 2. Large Scale production of Itaconic acid simulation using SuperPro Designer Software
Figure 3. shows that the total operating cost for itaconic acid production is 60,425,000USD (see Table 3).
Productivity is approximately 14 kg/h. The annual operating time setup to the Superpro Designer software is
7,920 hours. According to the calculations made in Table 3, the cost of production per kg is 555USD and the
cost of making a full container is 3 USD.

Figure 3. Operating costs per year.

The Company Enamine located in the United States sells 500mg of itanic acid at 25 USD. A desired profit
margin should be established and calculate the sale price taking into account the unit cost to determine the
cost of selling the product obtained through this simulation,. Figure 3 shows that the higher costs this process
is generating are in waste treatment with 41,721,000USD, 69% of all costs. To reduce them, an improvement
is proposed (see Figure 4).

Table 3 . Itaconic acid Productivity and Total USD calculated by SuperPro Designer

Total cost (USD) 60,425,000

Productivity (kg/h) 14

Hours per year 7920

Costs (USD/kg) 555

Costs USD/g 0.55

Cost (USD/container) 3

The proposed improvement is to use the water that evaporates in the first evaporator as a feed for the rotary
filter, but first it is passed through a condenser to go from steam to liquid and a recirculation of the second
outlet of the filter is also proposed to mix with the output of the industrial fermented. Recirculation turns out
to be an effective method for exploiting losses, SuperPro facilitates this process and the calculation of costs
including.

Figure 4. Improved Large Scale production of Itaconic acid simulation using SuperPro Designer Software
Table 4 shows that for the improved process the operating cost for product production is 18,781,000USD, the
productivity is 20.98 kg/h. As shown in Table 4, the cost of manufacturing per container of 500 mg of itanic
acid crystals is approximately 80 % cheaper those calculated without the improvement. Figure 5 illustrates
that the cost portion of waste treatment tents to be is 0. It was totally reduced. However, the Labor-
dependent and Facility-dependent percentage increased. The costs of the former did not increase but the
second if, before the improvement the cost was 7,769,000 and with the improvement the serious cost
8,712,000.

Figure 5. Executive Summary of The Improvement

Table 4.Improved Itaconic acid Productivity and Total USD calculated by SuperPro Designer

TotalCost 18,781,000
Productivity 20.98
Hours 7,920
Costs USD/kg 113.02
Costs USD/g 0.11
cost/container 1

CONCULSIONS
Simulation represents a great advantage for this was because it allows to know what is the best way to obtain
a substance of interest or how to improve a process, without having to do it in reality, this avoids many losses
and strengthens research to those areas that are difficult to study by different factors. Production of itaconic
acid using Aspergillus is somewhat difficult because of the little study in the area, the simulation represents a
solution to problems like this. It was successfully achieved to demonstrate how small modifications to the
process make a reduction in costs, in the simulation the cost of 41,721,000USD which belonged to the waste
treatment to zero and managed to reduce the operating costs was reduced. The process would have to be done
in reality to know that it is true that is raised and how much margin errothere is. Being an extracellular
metabolite where lysis was not needed, Biomass is a residual by-product to which an added value could be
given for decreased expenses and increased profits.

REFERENCES

Martínez Río, S. (2017). Diseño de un biorreactor para la producción biotecnológica de ácido itacónico (Tesis
de pregrado). Universidad Complutense De Madrid Facultad De Ciencias Químicas Departamento De
Ingeniería Química, Madrid, España.

García Henao, J. A. (2015). Evaluación de los efectos de la concentración de glicerol en la producción de

ácido itacónico a partir de AspergillusTerreus (Bachelor'sthesis, Uniandes) (Tesis de pregrado) Universidad
De Los Andes Facultad De Ingeniería Departamento De Ingeniería Quimica Bogotá, Colombia.

Rodríguez Rodríguez, E. (2018). Investigación de los métodos de obtención y polimerización del ácido
itacónico(Tesis de pregrado. Instituto politécnico nacional, Escuela superior de Ingeniería Química he
industrias extractivas, Ciudad de México.

Juy M. I., Orejas J. A., Lucca M. E.. Study of itaconic acid production by Aspergillus terrus MJL05 strain
with different variable. Rev. colomb. biotecnol  [Internet]. 2010 ;  Vol 12( 2 ): 187-193.

Petrides D, Carmichael D, Charles Siletti C and Koulouris A 2014 Biopharmaceutical Process Optimization
with Simulation and Scheduling Tools Bioengineering vol 1 pp 154-187