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MicroAlgal (MicroAlgae) Biorefinery Utilizing Dunaliella salina – Process

Modeling and Techno-Economic Assessment (TEA) using SuperPro Designer.

Preprint · August 2022

DOI: 10.13140/RG.2.2.11426.71365

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 MicroAlgal Biorefinery
Utilizing Dunaliella salina
Process Modeling and Evaluation using
SuperPro Designer®
by

Nikiforos Misailidis, Amir Mustafa, Rafael da Gama Ferreira, and Demetri Petrides
August 2022

This is the ReadMe file of a SuperPro Designer example that analyzes an integrated microalgal
biorefinery. Microalgae are grown in open solar ponds, utilizing the CO2 of the exhaust of a Combined
Heat and Power (CHP) gas turbine. After cell harvesting and disruption, heptane is used to extract the
non-polar components from the homogenized slurry. The non-polar components in the organic phase are
separated via membrane filtration to produce beta-carotene, the main product of the biorefinery, and free
fatty acids. The aqueous phase is processed to produce polar lipids, glycerol, and proteins. Ethanol is
used to precipitate part of the proteins, and acetone to precipitate the remaining proteins and
carbohydrates. All three solvents (heptane, ethanol, and acetone) are recovered and reused in the
process. The flowsheet of the process is appended to the bottom of this document. You may test-drive
this model by downloading the functional evaluation edition of SuperPro Designer from the downloads
page of our website (www.intelligen.com). All the files of this example can be found in the Examples \
Bio-Materials \ MicroAlgalBiorefinery folder. The default installation path of the SuperPro Designer
Examples folder follows below.
C:\ Users \ Public \ Public Documents \ Intelligen \ SuperPro Designer \ v# \ Process Library \ Examples

If you have any questions regarding this example or SuperPro Designer in general, please send an email
message to dpetrides@intelligen.com

INTELLIGEN, INC.
Simulation, Design, and Scheduling Tools
For the Process Manufacturing Industries
www.intelligen.com
Introduction

Microalgae species such as Dunaliella salina are well-known for their high content of valuable nutritional
and bioactive compounds such as carotenoids, glycerol, lipids, and proteins which are suitable for
nutraceutical & cosmeceutical applications. This example analyzes a sustainable process that utilizes
microalga D. salina and produces carotenoids, glycerol, polar lipids, and proteins

Microalgae are a promising feedstock as a source of biofuels, proteins and bioactive compounds that can
help address the problem of the growing demand for renewable sources in response to the increasing
world population and the need for sustainable energy and food sources. There is an economic need to
convert microalgal facilities into multiproduct biorefineries, moving away from microalgal processes solely
focused on biofuel production [1].

Figure 1. Application of microalgae-based products in various fields [2].

D. salina is a species of halophilic (well-adapted to unusually saline environments) unicellular microalgae

found particularly in hypersaline environments such as salt lakes and salt evaporation ponds. Known for its

2
antioxidant activity due to its ability to produce large amount of carotenoids, it is responsible for most of the
primary production in hypersaline environments worldwide and is also used in cosmetics and dietary
supplements [3]. D. salina is considered an extremophile, an organism that thrives in environments that
most organisms cannot tolerate due of its rare ability to survive in extreme salty habitats. In fact, D. salina
grows optimally at concentrations of 1.5–3.0 M NaCl, or about 3 to 6 times that of average seawater. Even
stranger, the pigment that makes it pink becomes increasingly concentrated when D. salina is exposed to
saltier environments, suggesting it may play a role in protecting the cell from high salt levels. However, D.
salina is mostly able to cope with extreme salinity for two different reasons. First, it has no cell wall (just a
malleable membrane), allowing it to expand and contract to maintain a habitable internal salt concentration.
Second, it is able to produce large amounts of glycerol to balance the osmotic pressure of the salt [4]. When
D. salina is grown at high salinity, the intracellular glycerol content exceeds 50%, which is sufficient to
balance out most of the required osmotic pressures. In this state, glycerol acts as a compatible solute,
protecting enzymes from both inactivation and inhibition.

In addition to this exceptional halotolerance of most Dunaliella species, Dunaliella acidophila can grow in
very acidic environment (pH 0–1). Dunaliella antarctica thrives in sub-zero temperatures and some strains
of D. salina can tolerate high light intensities. In addition, Dunaliella is more tolerant of fuel oil contamination
compared to other planktonic algae. As such, these organisms are unique in their ability to adapt to some
of the harshest conditions in global habitats [5].

Figure 2. Emanoil Constantin Teodorescu [6].

D. salina was first recognized in 1905 by Emanoil Constantin Teodorescu who was a Romanian botanist.
This alga is commonly found in natural marine habitats where it turns the water red in color. F.S Milko
reported in 1963 that Dunaliella contains high concentrations of β-carotene and later it was also
recommended as a commercial source of glycerol [7]. D. salina is a photosynthetic, unicellular, eukaryotic
organism, suitable for large-scale factory farming with simple and inexpensive culture. In addition, D. salina

3
does not have a rigid cell wall and has a single large chloroplast that can be transformed into its nucleus
and chloroplast simultaneously with the exogenous genes [8].

Figure 3. Lake Hillier, Australia. Pink Color due to the presence of Dunaliella salina microalgae [4].

4
 Figure 4. Colored scanning electron micrograph of Dunaliella salina. Magnification: x6000 when printed
10cm wide. Specimen courtesy of Mike Allen, Plymouth Marine Laboratory [9].

Figure 5 shows the ultrastructure of a D. salina cell; different components of the cell are identified with
different letters and the description of these components is given below.

(a) The cell membrane is mainly involved in cell growth and division, signal transduction, and changes in
osmotic pressure, which are closely related to the regulation of salt tolerance.

(b) The nuclear system is used for genome storage, gene expression and regulation. It may be hijacked
for recombinant protein expression as well.

(c) The cytoplasmic system, in which glycerol synthesis, nitrogen or phosphorus assimilation takes
place etc.

(d) The flagellar system is responsible for cell movement, cycle regulation, light capture, and other
processes.

(e) The chloroplast system is mainly involved in photosynthesis, lipid and carotenoid metabolism, and
carbon assimilation processes. Currently, it is also being developed as an expression system to produce
foreign proteins.

(f) Colonies of D. salina can be used in various applications, e.g. as biological fertilizer, animal feed,
biofuels, bioactive compounds, etc. [8]

5
 Figure 5. Ultrastructure of Dunaliella salina cell and its various components [8].

D. salina accumulates large amounts of β-carotene as droplets in the chloroplast to prevent chlorophyll
photo-damage when culture conditions include high light intensities, high temperature, high salinity, and
nutrient deficiency. β-carotene contents of up to 14% of dry weight have been reported for D. salina [5].
Apart from β-carotene, D. salina is also an important source of other lipidic components (10 – 25 % of dry
cell weight). Roughly 41% of the lipids present in D. salina are polar, suitable for nutraceutical &
cosmetical applications. Proteins can account for 10-40% of the total dry cell weight and are commonly
used for animal feed applications. In addition, carbohydrates represent up to 30% of the total dry cell
weight. Therefore, carotenoids, polar lipids, glycerol, proteins and carbohydrates may potentially be
recovered and purified from D. salina [1].

Large Scale Dunaliella salina Biorefinery

The first step before mass culture of D. salina in open ponds or bioreactors is to select strains that are best
suited to mass culture in terms of β-carotene content and growth rate. Only a few strains of D. salina can
accumulate large amounts of β-carotene (up to 10-14% of dry weight).

Since D. salina is an obligate photoautotroph, light is the only source of energy for its metabolism. In open
ponds, sunlight is the only light source, while in photobioreactors light can be provided either by using white
fluorescent lamps or by sunlight. Growth and carotenoid synthesis respond differently to different qualities
and quantities of light. In fact, carotenoid induction is wavelength independent, but strongly dependent on

6
light intensity. As for temperature, D. salina can thrive in a wide range of temperatures from below 0°C to
around 45°C. In laboratory cultures, the optimum temperature for D. salina growth is around 32°C with a
broad optimum between 25°C and 35°C. Due to technical limitations, the temperature in open ponds is not
controllable. On the other hand, temperatures around 40°C and more promote carotenoid induction but at
the same time slow down the growth rate. In addition, temperatures above 40°C cause dramatic leakage
of glycerol into the medium, which can serve as an organic carbon source for bacteria and filamentous fungi
which may then become dominant.

Dunaliella species have a wide range of pH tolerance from 0 to 11, but the optimal pH for D. salina is
between 9 and 11. There is a risk of precipitation by multiple calcium salts and flocculation of the algal
biomass at higher pH, especially when the Ca2+ concentration is high, which are conditions normally found
in many natural water sources. In some open ponds and photobioreactors, where the main source of
inorganic carbon is bicarbonate ion, the pH is only controlled by the addition of HCl. Since Dunaliella is
photoautotrophic, it can only use carbon dioxide and bicarbonate as inorganic carbon sources. The lack of
a suitable inorganic carbon source is the most common growth limiting factor under common culture
conditions (high salinity, elevated pH and high temperature). Typically, gas bubblers inject small bubbles
of CO2 into the culture. Alternatively, NaHCO3 can be used as a carbon source for good growth between
pH 7.5 and 9.5; However, the algae will grow well up to a pH of 11 when a higher initial concentration of
bicarbonate is provided. This is because at higher pH, the supply of soluble carbon dioxide becomes
restricted, such that more bicarbonate is needed. The best source of nitrogen for D. salina is nitrate. In
practice, 5 mmol/L of NaNO3 or KNO3 are added to the medium for optimal growth. On the other hand,
nitrate limitation is one of the most common ways for reducing the growth rate, which leads to the induction
of carotenoid production. However, prolonged nitrogen limitation in the culture can eventually lead to high
cell mortality as well as a severe reduction of carotenoids per unit culture volume. Other sources of nitrogen
such as ammonium salts and urea are not suitable, because under certain conditions they can kill the algae.
Phosphorus in the forms of KH2PO4 or NaH2PO4 gives the best results. Dunaliella also requires high
concentrations (approximately 2 mmol/L) of sulfate for maximum growth, but this is rarely added to
commercial pond medium as natural water sources such as seawater or tap water contain much higher
contents of sulfate, around 30 mmol/L). Other elements required for good growth of D. salina include K+,
Ca2+, Mg2+, Cl-, Na+, chelated iron and trace elements. The ratios of Mg2+: Ca2+ and Cl--: SO42-) in the medium
can also affect both growth and carotene synthesis. However, in most cases, there is no need to add these
elements to the medium if it is composed of technical grade salt or seawater [5] [7].

Clearly, D. salina cells consists of several valuable components at variable concentrations depending on
the medium concentration and culture conditions. Carotenoids are the main high value product of such a
facility. Carotenoids are antioxidants, with cancer fighting properties. When consumed by humans, they are
converted to vitamin A, which helps to improve vision and normal growth and development. Carotenoids
also have anti-inflammatory and immune system benefits. The growth of the carotenoid market is attributed

7
to their increasing applications in pharmaceuticals, cosmetics, food, and animal feed. The global
carotenoids market is expected to grow from $2.34 billion in 2017 to $3.59 billion in 2025, representing a
CARG of 5.5% [10]. The beta carotene market size was $520 million in 2020, and is estimated to grow at
a CARG of larger than 6% until 2027 [11].

Process Description
A conceptual micro-algal biorefinery process was modeled and economically evaluated using SuperPro
Designer to estimate the expected material requirements, process equipment, utilities consumption and
ultimately production costs. For reporting and analysis purposes, the process has been divided into five
sections:

➢ Media Preparation and Culture Ponds (black icons)

➢ Combined Heat and Power (dark red icons)
➢ Harvesting (blue icons)
➢ Organic Phase Processing (green icons)
➢ Aqueous Phase Processing (orange icons)
➢ Acetone Extraction (turquoise icons)
➢ Solvents Recovery and Reuse (violet icons)

Flowsheet sections in SuperPro are simply sets of related unit procedures (processing steps). For
information on how to specify flowsheet sections and edit their properties, please consult Chapter 8.1 of
the SuperPro manual. The contents of each of this example’s flowsheet sections are described in greater
detail below. The process will be easier to follow if you open this example within SuperPro and view the
flowsheet while reading the description of the process.

Media Preparation and Culture Ponds

This section includes media preparation and microalgae culture ponds. The media prepared consist of salts.
As explained earlier, high salinity media offers several advantages including higher carotenoid content and
reduced risk of contamination. The basic carbon source for microalgae growth is the Carbon Dioxide (CO 2)
from the exhaust gases of a Combined Heat and Power (CHP) system, which is a section on its own and
which is later described.

The process begins with the solar ponds in procedure P-1, which operate in continuous mode. The high
salinity media are prepared by mixing a recycling stream, which will be later analyzed, with seawater in a
custom mixer (P-2). Potassium Nitrate (KNO3) is stored in hopper P-4 and mixed in the media via the P-5
custom mixer at a concentration of about 1.63 g/L. Sodium Bicarbonate is stored in hopper P-6 and mixed
into the media via the P-7 custom mixer to a final concentration of 4.2 g/L. Finally, the salinity of the

8
media is increased by adding Sodium Chloride, which is stored in P-8 and added via the P-9 custom
mixer to a final concentration of 100 g/L. The prepared media are continuously fed to the solar ponds P-1.
The solar ponds operation includes three reactions, representing the dissociation of Potassium Nitrate,
the algae growth and CO2 neutralization. The reactions have the following stoichiometries and
conversions:

Solar Pond Reactions – Mass or Molar Stoichiometries Conversion

KNO3 Dissociation (molar):

1 KNO3 + 1 Water → 1 HNO3 + 1 KOH 100%

Algae Growth (mass):

44 CO2 + 5 HNO3 + 2 MgSO4 + 1 NaCl + 16 Water → 16% (resulting

23.5 D.salina + 44.5 O2 from the target
concentration
Target concentration: 1 g/L of D.salina specification)

Neutralization (molar):

1 CO2 + 2 KOH → 1K2CO3 + 1 Water 100%

Combined Heat and Power (CHP)

As explained above, there is a gas turbine system in the facility, which provides the necessary electricity
and heat to run the plant, and supplies the solar ponds with CO2, which is the carbon source for the
growth of the microorganisms. Air is compressed in the gas turbine P-10, then mixed with natural gas.
The mixture is combusted, increasing the pressure of the gases, and then expanded through the turbine
to generate electricity. 2 MT/h of natural gas is consumed to generate around 8 MW of electricity,
corresponding to an electrical efficiency of about 30%. The exhaust gases exit at a temperature of slightly
above 900 °C and a CO2 content of 8.7% w/w and are used to generate around 20 MT/h of 5 bar steam in
P-11. The exhaust gases from P-11 are used to generate 60 MT/h of hot water in P-12, then mixed with
fresh air in P-13 and bubbled in the solar ponds for carbonation and algal growth. The overall heat input
of about 26.3 MW of natural gas is converted into electricity (30%), steam (53%) and hot water (11.5%).

Harvesting

After the solar ponds, the microalgae slurry is harvested using a decanter centrifuge (P-14). The
supernatant of the centrifuge is split in P-15. 90% of the solution is recycled back to P-2 to be mixed with
fresh sea water. The remaining 10% is sent to wastewater treatment. The concentrated algae in the
underflow of the first centrifuge (stream S-102) is mixed with water in a custom mixer (P-16) and passed
through another centrifuge (P-17). This washing step significantly reduces the amount of salt in the

9
biomass. The slurry is then sent to a high-pressure homogenizer (P-18) to disrupt the cellular structure.
The homogenization reaction has the following stoichiometry:

Homogenization Reaction – Mass Stoichiometry Conversion

100 D.salina → 5 β-carotene + 18 Carbohydrates + 2 FFA + 26 Glycerol + 10 100%

Lipids + 7 Polar Lipids + 24 Proteins + Salts

The homogenate is then mixed with heptane in a custom mixer (P-19) and enters a decanter separator
(P-20). Water is a polar solvent and as such not miscible with the non-polar heptane, so that the solution
forms two phases. The separation in the decanting unit is based on partition coefficients. Heptane was
set as the light organic phase, while water was set as the heavy phase. The homogenized components of
the algae are separated between the two phases according to the polarity of their molecules. The partition
coefficients of each component were set accordingly. Carotenoids, free fatty acids, and triglyceride lipids
partition in the heptane phase, while salts and polar lipids remain in the aqueous phase.

Organic Phase Processing

The heptane organic phase is sent to the “Organic Phase Processing” section. The first step involves a
neutralization reactor (P-21). The neutralization takes place with a 10% potassium hydroxide aqueous
solution, which is prepared in a custom mixer (P-22). In the neutralization reactor the lipids are hydrolyzed
and saponified. The neutralization reactions have the following molar stoichiometries:

Neutralization Reactions – Molar Stoichiometries Conversion

Lipids Hydrolysis:

1 Lipid + 3 Water → 3 FFA + 1 Glycerol 100%

FFA saponification:

1 FFA + 1 KOH → 1 Soaps + 1 Water 30%

The saponified mixture enters another decanter (P-23), which separates the two phases. The soaps in
the aqueous phase are removed using a filter (P-24), while the remainder of the solution (stream S-185)
is sent to the aqueous phase processing section. The light organic phase contains the carotenoids and
the smaller free fatty acid molecules. These are separated based on their size differences using a
nanofiltration unit (P-25).

The retentate of the nanofiltration unit (stream S-136) which contains most of the carotenoids is sent to a
thin film evaporator (P-26) to remove the heptane solvent. The evaporator operates under vacuum to

10
remove heptane at reduced temperatures, thus avoiding damage to the carotenoid molecules. The
product stream is then flashed in P-27 at a lower pressure to evaporate the residual heptane. The liquid
of the flash tank is pumped in P-28 and cooled down to 30 °C in P-29 against cooling water. This stream
contains the carotenoids and is the main product of the algae biorefinery. The vapors from the thin film
evaporator and the flash tank, containing heptane, are mixed in P-30, condensed against cooling water in
a condenser (P-31) which operates at 0.2 bar pressure, and pumped in P-32. The liquid heptane is mixed
with another heptane stream (which is later analyzed) in P-40 and recycled back to the “Harvesting”
section.

The filtrate of the nanofiltration (stream S-135), which mostly contains free fatty acids, is sent to another
thin film evaporator (P-34), which evaporates the heptane solvent. The final traces of heptane are
evaporated in a subsequent flash tank (P-34), which operates at a lower pressure. The free fatty acids
are then pumped in P-35 and cooled down in P-36 against cooling water. The heptane vapors from
evaporator P-34 and flash tank P-35 are combined in P-37 and condensed in P-38 against cooling water.
The liquid heptane is then pumped into P-39 and sent to the P-40 mixer as mentioned above.

As a result, the Organic Phase Processing section separates and purifies two products: carotenoids and
free fatty acids.

Aqueous Phase Processing

The heavy phase of the P-20 decanter (stream S-126) is sent to the Aqueous Phase Processing section
where the aqueous solution is mixed with 94% w/w ethanol solution in a custom mixer (P-41) up to a final
ethanol content of around 30%. The high ethanol concentration precipitates most of the proteins in the
reactor that follows (P-42). The precipitation reaction has the following mass stoichiometry:

Protein Precipitation – Mass Stoichiometry Conversion

100 Protein → 100 Protein(s) 70%

The suspension is then mixed in P-43 with the filtrate of the soap filtration (stream S-185) from the
Organic Phase Processing, and a recycled stream (S-168) which will be later analyzed. The solution is
centrifuged in P-44, which separates the precipitated proteins. The concentrated proteins are further
drained and washed with water in a belt filter (P-45). The filtrate (stream S-168) is recycled back to the
mixer P-43. In addition to reducing the moisture content of the proteins stream prior to the dryers, the
washing of the belt filter recovers and recycles most of the ethanol contained in the stream. The protein
solids (stream S134) are then mixed in a mixer (P-46) with another protein-rich stream which will be later
analyzed and sent to a rotary dryer (P-48). The protein-rich stream is dried with hot air and cooled with

11
ambient air in the two sections of the rotary dryer. The final dried material has a water content of 10%
w/w. Proteins are the first product obtained from the aqueous phase. NOTE - Detailed information on how
to initialize rotary dryers that include both drying and cooling sections is provided in the Miscellaneous
Modeling Tips section close to the bottom of this document.

The hydroethanolic phase in the supernatant of the centrifuge (stream S-128) contains glycerol,
carbohydrates, some soluble proteins, polar lipids, and salts. These are initially separated by a suitable
membrane filter (P-49) according to their molecular sizes. The larger molecules of soluble proteins, large
carbohydrates, and polar lipids remain in the retentate, while the salts, glycerol and small carbohydrates
pass through the membrane. The retentate is washed with water in a mixer (P-50) and passed through
another membrane filter (P-51). This step further purifies the large molecules by reducing the amount of
small molecules present. The filtrates of the two membrane filters (streams S-198 and S-199) are
combined in P-52 and fed to a multi-effect evaporator (P-53), which concentrates the glycerol solution
using steam. All the ethanol and most of the water are evaporated. The vapors of the last stage are
condensed in a condenser (P-54) against cooling water, pumped (P-55) and sent to a mixer (P-56) in the
Solvents Recovery and Reuse section to recover the ethanol.

The retentate of the P-51 membrane filter (stream S-113) is concentrated by a multi-effect (two-stage)
evaporator (P-57) using steam. The vapors from the second stage are condensed in a condenser (P-58)
against cooling water and pumped in P-59. They are mixed with the liquid from the first stage in mixer P-
56 to recover and reuse the contained ethanol. The concentrated solution (stream S-125) contains
around 92% water. It is cooled in P-60 and pumped by P-61 to a mixed bed ion exchange column (P-62),
which removes the salt ions from the solution. The column is regenerated and washed using water from
flow distributor P-63, hydrochloric acid 7% w/w prepared in the custom mixer P-65, and sodium hydroxide
solution 4% w/w prepared by P-64. The mixed bed ion exchange procedure operates in batch (cyclical)
mode and includes the following operations:

- Load. The product solution is loaded onto the column and the purified product is collected in the
outlet.
- Wash Product Solution. The product loading stops, and the remaining product solution in the
resin is washed out with some water. The column outlet during this wash step is combined with
the product stream.
- Wash Column. The column is washed further, with the wash water ending up in the IX waste.
- Regenerate 1. The resin is regenerated using 7% hydrochloric acid. The outlet during this step is
sent to the IX waste stream.
- Regenerate 2. The resin is regenerated using 4% sodium hydroxide. The outlet during this step
is sent to the IX waste stream.
- Wash 1. The regenerated column is washed with water to remove the regeneration solutions and
prepare the resin for the next cycle.

12
There are two mixed bed ion exchangers operating in staggered mode. The cycle time was set equal to
the process time of the “Load” operation in a way that there is always an ion exchanger in load mode,
while the other unit is being regenerated. For more information on the use of staggered equipment,
please consult the Industrial Enzymes production example.

Acetone Extraction

The product solution from P-62 is free of ions and contains soluble proteins, carbohydrates, and polar
lipids. An additional precipitation step is performed with acetone to remove the polar lipids. Acetone is
added via a custom mixer (P-66) up to a final acetone composition of 50% w/w and the solution is sent to
a precipitation vessel (P-67). Most of the proteins and carbohydrates precipitate. The precipitation
reactions have the following mass stoichiometries:

Acetone Precipitation – Mass Stoichiometries Conversion

Protein Precipitation:

100 Protein → 100 Protein(s) 95%

Carbohydrates Precipitation:

100 Carbohydrates → 100 Carbohydrates(s) 95%

The precipitated solids are separated using a disk-stack centrifuge (P-68) to a final solids concentration of
400 g/L and further concentrated and washed using a belt filter (P-69). The washed solids (stream S-178)
are combined with the ethanol precipitated proteins (in P-46) and sent to the proteins dryer (P-47). The
supernatant of the P-68 disk-stack centrifuge (stream S-123) is concentrated in a two-stage evaporator
(P-70). The vapors from the last evaporation stage are condensed (in P-71) against cooling water and
pumped in P-72 to the P-73 mixer in the Solvents Recycle and Reuse section, to recover and reuse the
acetone. The concentrated product solution contains the polar lipids, which are considered a co-product.

Solvents Recovery and Reuse

The process uses three solvents: heptane, ethyl alcohol and acetone. Each of these solvents is
recovered and reused in the process.

Heptane Recycling
Heptane is immiscible in water. In the process it is added in the custom mixer P-19 and separated from
the water solution in the decanters P-20 and P-23. The evaporated and condensed heptane streams are
combined in P-40 and recycled back to the process via a make-up mixer (P-74) and a custom splitter (P-

13
75). The mixer P-74 adds some make up heptane, while the custom splitter P-75 allows the process to
pull out the required amount of heptane. The rest is purged (stream S-209).

Ethyl Alcohol Recycling

Ethyl alcohol is added in the process via the P-41 custom mixer to precipitate proteins. The added
ethanolic stream has a concentration of around 94% w/w. The alcohol is then recovered in the vapor
streams of the P-53 and P-57 evaporators, which are combined in P-56. The resulting water / ethanol
mixture (stream S-148) is preheated to a final temperature of about 90 °C and fed to a distillation column
(P-77). The VLE calculations for the rigorous distillation column were performed based on “Modified
Raoult’s Law” using “Wilson” binary coefficients. For additional information on the use of Rigorous VLE
modelling, please consult the Isobutanol example in the Biofuels folder. The ethyl alcohol is taken from
the distillate at a composition just below the azeotropic mixture between ethanol and water, at around
94% w/w. Ethyl alcohol is then cooled down to 35 °C (in P-78) against cooling water and recycled back to
the process via the P-79 make-up mixer and the P-80 custom splitter. The P-79 mixer adds some make-
up ethanol, while the custom splitter P-80 operates in pull out mode supplying the process with the
needed amount of ethanol.

The stillage in the bottom of the column contains water. It is used to preheat the column feed solution in a
heat exchanger (P-76). It is then further cooled down in P-81 against cooling water and distributed back
to the process via the custom splitter P-82, which operates in pull out mode, and the flow distributor P-83.
The users of this water stream are the two belt filters (P-45 and P-69) and the P-50 mixer. In the belt
filters, water washes the cake replacing the ethanol and acetone solvents. In the P-50 mixer, it washes
the retentate to reduce the amount of low molecular weight contaminants.

Acetone Recycling
Acetone is added to the process via the P-66 custom mixer to precipitate proteins and carbohydrates.
The added stream has a concentration of around 98.5% w/w acetone. It is then recovered in the vapors of
the P-70 evaporator and in the cake wash of the P-69 belt filter (stream S-190). The two acetone streams
are combined in P-73. At this stage the acetone forms a dilute solution with water, containing around 40%
w/w acetone. This solution is preheated to a final temperature of around 55 °C (in P-84) and fed to a
distillation column (P-85). This distillation column was also modeled using a rigorous distillation
procedure, and the selected phase equilibrium model was set to be based on “Modified Raoult’s Law”
using “Wilson” binary coefficients. Acetone exits in the distillate with a composition of around 98.5% w/w.
The acetone is then cooled down to 35 °C (in P-86) against cooling water and recycled back to the
process via the P-87 make-up mixer and the P-88 custom splitter. The mixer P-87 adds some make-up
acetone, while the custom splitter P-88 operates in pull out mode supplying the process with the needed
amount of acetone. The stillage from the column is used to preheat the column feed stream (in P-84), and
then sent to wastewater treatment.

14
Economic Evaluation

This plant produces around 170 MT of β-carotene per year. Various assumptions were made for the costs
of raw materials, heat transfer agents, wastewater treatment, equipment purchase costs, labor, etc. In
addition to mass and energy balances, SuperPro Designer calculates the capital (CAPEX) and operating
expenses (OPEX), production cost and profitability of the project. The results can be found in the
Economic Evaluation (EER), Cash Flows Analysis (CFR), Itemized Cost (ICR) and Excel Custom reports.
In addition to β-carotene, the algal biorefinery produces several co-products with variable purities and
yields. Table 1 summarizes the various products produced by this process and their yields on recovered
dried algae. Glycerol is the component with the highest concentration in the algae cells. However, it is
recovered at a very low purity, since most of the water-soluble impurities (salts carbohydrates etc.), end
up in this stream. Proteins is the second highest recovered co-product in terms of volume. Beta carotene
has the lowest amount yield, but it is the main product of the process because of its high selling price.

Table 1. Flowrates, purities, and yields of the algal biorefinery process.

Total Purity on Yield of pure
Pure
Stream DS (%) substances on
component
Flow (kg/h) Recovered
flow (kg /h)
Algae DS (%)
Beta-Carotene 34.1 62.9 21.5 4.80
Proteins 150.0 78.0 105.9 23.64
Polar Lipids 67.3 76.2 28.5 6.36
Free Fatty Acids 34.4 98.5 33.9 7.57
Glycerol 303.7 44.0 114.0 25.45

Table 2, which was extracted from the EER, provides information on equipment sizes and purchase
costs. The SuperPro Designer built-in cost models were used for estimating the purchase cost of most
equipment items. User-defined cost models were used for estimating the purchase cost of tanks, reactors,
solar ponds, centrifuges, and decanters. The specification of user-defined cost models is explained in
detail in the manual and in the ReadMe file of the Corn Refinery example of SuperPro. Table 3, which
was also extracted from the EER, provides an estimate of the Direct Fixed Capital Cost, which is around
$144 million for a plant of this capacity.

15
2. MAJOR EQUIPMENT SPECIFICATION AND FOB COST (2022
prices)
Quantity/
Standby/ Name Description Unit Cost ($) Cost ($)
Staggered
1/0/0 DC-101 Decanter Centrifuge 5,770,000 5,770,000
Throughput = 444828.66 L/h
1/0/0 GT-101 Gas Turbine-Generator 4,431,000 4,431,000
Electric Power = 8019.23 kW
1/0/0 DS-103 Disk-Stack Centrifuge 1,287,000 1,287,000
Throughput = 9778.47 L/h
1/0/0 DS-104 Disk-Stack Centrifuge 772,000 772,000
Throughput = 3791.42 L/h
1/0/0 EV-101 Multi-Effect Evaporator 715,000 715,000
Mean Heat Transfer Area = 85.28
1/0/0 C-106 Rigorous Distillation Column 700,000 700,000
m2
Column Volume = 6773.66 L
6/0/0 RP-101 Raceway Pond 621,000 3,726,000
Vessel Volume = 43075.26 m3
1/0/0 DC-102 Decanter Centrifuge 578,000 578,000
Throughput = 27725.55 L/h
1/0/0 V-101 Neutralizer 504,000 504,000
Vessel Volume = 321.17 m3
1/0/0 C-104 Rigorous Distillation Column 500,000 500,000
Column Volume = 962.38 L
1/0/0 HX-102 Heat Exchanger 440,000 440,000
Heat Exchange Area = 692.30 m2
1/0/0 HX-101 Heat Exchanger 302,000 302,000
Heat Exchange Area = 369.52 m2
1/0/0 EV-103 Multi-Effect Evaporator 299,000 299,000
Mean Heat Transfer Area = 12.14
1/0/0 TFE-102 Thin Film Evaporator 287,000 287,000
m2
Film Area = 6.37 m2
1/0/0 BF-102 Belt Filter 283,000 283,000
Belt Width = 0.20 m
1/0/0 BF-101 Belt Filter 282,000 282,000
Belt Width = 0.25 m
1/0/0 EV-102 Multi-Effect Evaporator 253,000 253,000
Mean Heat Transfer Area = 4.08 m2
1/0/0 V-102 Decanter Tank 197,000 197,000
Vessel Volume = 8.86 m3
1/0/0 V-103 Decanter Tank 192,000 192,000
Vessel Volume = 8.53 m3
1/0/0 R-101 Stirred Reactor 191,000 191,000
Vessel Volume = 9.34 m3
1/0/0 R-102 Stirred Reactor 163,000 163,000
Vessel Volume = 21.02 m3
1/0/0 HP-103 Hopper 160,000 160,000

16
 Vessel Volume = 21955.72 L
1/0/0 TFE-101 Thin Film Evaporator 135,000 135,000
Film Area = 1.58 m2
1/0/1 INX-102 Ion Exchanger 133,000 266,000
Column Volume = 2.26 m3
1/0/0 RDR-102 Rotary Dryer 120,000 120,000
Drying Area = 15.23 m2
1/0/0 HP-101 Hopper 110,000 110,000
Vessel Volume = 11797.03 L
1/0/0 HX-109 Heat Exchanger 98,000 98,000
Heat Exchange Area = 56.45 m2
1/0/0 NF-101 Reverse Osmosis Filter 78,000 78,000
Membrane Area = 432.92 m2
1/0/0 PM-109 Centrifugal Pump 74,000 74,000
Pump Power = 18.29 kW
1/0/0 NF-102 Reverse Osmosis Filter 71,000 71,000
Membrane Area = 384.16 m2
1/0/0 HG-101 Homogenizer 70,000 70,000
Rated Throughput = 5305.80 L/h
1/0/0 NF-103 Reverse Osmosis Filter 57,000 57,000
Membrane Area = 298.64 m2
1/0/0 HX-108 Heat Exchanger 56,000 56,000
Heat Exchange Area = 22.40 m2
1/0/0 HX-105 Condenser 54,000 54,000
Condensation Area = 162.82 m2
1/0/0 HP-102 Hopper 51,000 51,000
Vessel Volume = 3251.91 L
1/0/0 HX-106 Condenser 43,000 43,000
Condensation Area = 86.87 m2
1/0/0 HX-113 Condenser 38,000 38,000
Condensation Area = 9.35 m2
1/0/0 HX-103 Condenser 38,000 38,000
Condensation Area = 8.49 m2
1/0/0 HX-104 Condenser 38,000 38,000
Condensation Area = 52.87 m2
1/0/0 HX-107 Heat Exchanger 30,000 30,000
Heat Exchange Area = 8.07 m2
1/0/0 MF-101 Microfilter 29,000 29,000
Membrane Area = 1.07 m2
1/0/0 HX-110 Heat Exchanger 16,000 16,000
Heat Exchange Area = 2.72 m2
1/0/0 PM-102 Centrifugal Pump 13,000 13,000
Pump Power = 0.34 kW
1/0/0 PM-104 Centrifugal Pump 12,000 12,000
Pump Power = 0.27 kW
1/0/0 PM-106 Centrifugal Pump 10,000 10,000
Pump Power = 0.00 kW
1/0/0 PM-108 Centrifugal Pump 10,000 10,000
Pump Power = 0.01 kW
1/0/0 PM-107 Centrifugal Pump 10,000 10,000
Pump Power = 0.07 kW
1/0/0 PM-101 Centrifugal Pump 10,000 10,000
Pump Power = 0.00 kW

17
1/0/0 PM-103 Centrifugal Pump 10,000 10,000
Pump Power = 0.08 kW
1/0/0 PM-105 Centrifugal Pump 10,000 10,000
Pump Power = 0.06 kW
1/0/0 HX-112 Heat Exchanger 10,000 10,000
Heat Exchange Area = 1.26 m2
1/0/0 HX-115 Heat Exchanger 9,000 9,000
Heat Exchange Area = 0.08 m2
1/0/0 HX-114 Heat Exchanger 9,000 9,000
Heat Exchange Area = 0.08 m2
1/0/0 HX-111 Heat Exchanger 9,000 9,000
Heat Exchange Area = 0.21 m2
1/0/0 V-104 Flash Drum 2,000 2,000
Vessel Volume = 57.82 L
1/0/0 V-105 Flash Drum 2,000 2,000
Vessel Volume = 65.87 L
Unlisted Equipment 5,907,000

Auxiliary
TOTAL 29,536,000
Equipment

3. FIXED CAPITAL ESTIMATE SUMMARY (2022 prices in $)

3A. Total Plant Direct Cost (TPDC) (physical cost)

1. Equipment Purchase Cost 29,536,000
2. Installation 12,628,000
3. Process Piping 6,680,000
4. Instrumentation 6,937,000
5. Insulation 886,000
6. Electrical 2,954,000
7. Buildings 7,271,000
8. Yard Improvement 4,430,000
9. Auxiliary Facilities 6,937,000
TPDC 78,259,000

3B. Total Plant Indirect Cost (TPIC)

10. Engineering 19,565,000
11. Construction 27,391,000
TPIC 46,955,000

3C. Total Plant Cost (TPC = TPDC+TPIC)

TPC 125,214,000

3D. Contractor's Fee & Contingency (CFC)

12. Contractor's Fee 6,261,000
13. Contingency 12,521,000
CFC = 12+13 18,782,000

3E. Direct Fixed Capital Cost (DFC = TPC+CFC)

DFC 143,996,000

18
Tables 4a,b,c provide information on the assumed unit costs and the calculated annual amounts and
costs for a) raw materials, b) labor and c) utilities. The total annual cost of raw materials is around $12.7
million. According to Table 3b the cost of labor is approximately $10 million per year. Table 5c displays
the utilities costs, which were estimated to be about $6.3 million per year. Note that the generated steam
and power from the CHP unit are calculated later as revenues.

4a. MATERIALS COST - PROCESS SUMMARY

Unit Cost Annual Annual Cost

Bulk Material %
($) Amount ($)
Acetone 1.00 39,600kg 39,600 0.31
Air 0.00 1,283,892,269kg 0 0.00
Ethyl Alcohol 1.00 39,600kg 39,600 0.31
HCl(aq) 0.15 569,070kg 85,361 0.67
Heptane 0.50 15,840kg 7,920 0.06
KNO3 0.40 5,956,318kg 2,382,527 18.78
KOH(aq) 0.40 21,929kg 8,772 0.07
NaCl 0.00 22,170,887kg 0 0.00
NaHCO3 0.40 1,641,889kg 656,756 5.18
NaOH(aq) 0.40 315,293kg 126,117 0.99
NG 0.43 15,840,000kg 6,791,486 53.53
Sea Water 0.00 533,432,235kg 0 0.00
Water 3.00 849,553MT 2,548,658 20.09
TOTAL 12,686,797 100.00
NOTE: Bulk material consumption amount includes material used as:
- Raw Material
- Cleaning Agent
- Heat Transfer

4b. LABOR COST - PROCESS SUMMARY

Unit Cost Annual Amount Annual Cost

Labor Type %
($/h) (h) ($)
Operator 69.00 143,006 9,867,434 100.00
TOTAL 143,006 9,867,434 100.00

4c. UTILITIES COST (2022 prices) - PROCESS SUMMARY

Unit Cost Annual Ref. Annual Cost

Utility %
($) Amount Units ($)
Std Power 0.10 14,054,612 kW-h 1,405,461 22.18
Steam 30.00 139,862 MT 4,195,870 66.22
Cooling Water 0.05 14,707,221 MT 735,361 11.60
TOTAL 6,336,693 100.00

19
Figure 6 shows a breakdown of the total annual operating costs. Clearly the facility-dependent cost
(depreciation of the capital investment, maintenance, etc.), raw materials and labor have the highest
contribution to the total annual operating costs, accounting for 45%, 21% and 16%, respectively.

Figure 6. Annual Operating Cost Breakdown (%).

As discussed, the analyzes algal biorefinery generates several co-products with variable yields and
selling prices. In addition, the CHP section produces energy in various forms. Table 5, which was
extracted by the EER, summarizes all the annual production rates of all the coproducts of the algal
biorefinery, their assumed selling prices, and their expected annual revenues. Clearly beta carotene is the
predominant revenue source, followed by electricity and steam, contributing around $59, $6.3 and $4.8
million of annual revenues. However, part of the produced electricity and most of steam is used to cover
the needs of the plant itself, as summarized in Table 4c above. Proteins and polar lipids are considerable
material revenue streams that contribute around $4.8 and $3.7 million per year with the assumed selling
prices. Further processing and purification of some co-products might increase their selling prices and
ultimately improve the economic bottom line of the plant.

20
Table 5. Annual production rates, selling prices, and annual revenues of material and energy co-
products of the algal biorefinery process.
Annual Revenue Selling Price
Annual Revenue ($)
Rate
Beta Carotene 169,935 kg $350/kg 59,477,387
Proteins 1,192,542 kg $4/kg 4,770,167
Polar Lipids 533,398 kg $7/kg 3,733,784
Free Fatty Acids 272,548 kg $2/kg 545,096
Glycerol 902,500 kg $0.5kg 451,250
Utilities
Steam 160,776 MT $30/MT 4,823,280
Hot Water 475,200 MT $0.5/MT 237,600
Electricity 63,512,312 kWh $0.1/kWh 6,351,231

Total Revenues 80,389,880

Finally, Table 6 displays the executive summary. The total CAPEX required was estimated to be about
$154 million. For a selling price of $350/kg of β-carotene produced, the expected payback time is around
5.5 years.

6. EXECUTIVE SUMMARY (2022 prices)

Total Capital Investment 154,110,000 $

Capital Investment Charged to This Project 154,110,000 $
Operating Cost 60,934,000 $/yr
Main Revenue 59,477,000 $/yr
Other Revenues 20,912,493 $/yr
Total Revenues 80,390,000 $/yr
Cost Basis Annual Rate 169,935 kg MP/yr
Unit Production Cost 358.57 $/kg MP
Net Unit Production Cost 358.57 $/kg MP
Unit Production Revenue 473.06 $/kg MP
Gross Margin 24.20 %
Return On Investment 18.34 %
Payback Time 5.45 years
IRR (After Taxes) 11.60 %
NPV (at 7.0% Interest) 74,344,000 $
MP = Flow of Component 'B-Carotene' in Stream 'β-carotene'

21
Miscellaneous Modeling Tips

Initializing a Rotary Dryer in SuperPro v13

In SuperPro Designer v13, the rotary, spray and fluidized bed dryers can optionally include a section for
cooling the dried product. Figure 7a displays the “Operating Conditions” tab of the P-48 rotary dryer unit
that includes both drying and cooling sections (note that both the “Use Hot Gas” and the “Use Cold Gas”
options are enabled). The “Use Cold Gas” box brings up the “Post-Drying Cooling” tab through which you
specify parameters for product cooling. When that option is activated, a second inlet air/gas stream needs
to be attached to the “Secondary Gas In” inlet port of the unit along with a second outlet air/gas stream.

The option “Preheat the inlet Gas” allows the user to specify the desired hot air/gas temperature, which is
accomplished by heating the inlet air/gas stream with a heating agent. Another new feature is the ability
to specify entrainment of drying solids to the outlet air stream. In fact, each section can have its own
solids entrainment parameter.

Figures 7b & c display the “Main Drying” and “Post-Drying Cooling” tabs. The user can specify the
volatiles content (LOD) and the product temperature after each section.

22
b

23
Figure 7. The Operating conditions, Main Drying and Post-Drying Cooling interfaces of the rotary dryer in
SuperPro v13.

Summary

The objective of this example was to analyze an algal biorefinery process in SuperPro Designer that is
easy to understand and follow. As indicated in the preceding analysis, a plant with a capacity of around
170 metric tons of β-carotene per year requires a total CAPEX of around $154 million and annual
operating expenditures (including depreciation) of around $61 million. The predominant cost is the facility-
dependent cost, followed by the cost of raw materials. Assuming a selling price of $350/kg of β-carotene,
the payback time for such an investment was estimated to be around 5.5 years.

References

[1] J. Monte et al., “Biorefinery of Dunaliella salina: Sustainable recovery of carotenoids, polar lipids and
glycerol,” Bioresour. Technol., vol. 297, p. 122509, Feb. 2020, doi: 10.1016/j.biortech.2019.122509.
[2] K. N. Chidambara Murthy, “Production of Beta-Carotene from Cultured Dunaliella Sp. and Evaluation
of Biological Activities.” CENTRAL FOOD TECHNOLOGICAL RESEARCH INSTITUTE, Dec. 2005.
[Online]. Available: https://www.researchgate.net/publication/49177875_Production_Of_Beta-
Carotene_from_Cultured_Dunaliella_Sp_and_Evaluation_Of_Biological_Activities
[3] “Dunaliella salina,” Wikipedia. Mar. 06, 2022. Accessed: May 10, 2022. [Online]. Available:
https://en.wikipedia.org/w/index.php?title=Dunaliella_salina&oldid=1075535561
[4] A. R. Supply, “Dunaliella salina: the alga that’s always pretty in pink,” Algae Research Supply.
https://algaeresearchsupply.com/pages/dundunaliella-salina-the-algae-that-s-always-pretty-in-pink
(accessed May 10, 2022).
[5] A. Hosseini Tafreshi and M. Shariati, “Dunaliella biotechnology: methods and applications,” J. Appl.
Microbiol., vol. 107, no. 1, pp. 14–35, Jul. 2009, doi: 10.1111/j.1365-2672.2009.04153.x.
[6] “Emanoil C. Teodorescu,” Wikipedia. Apr. 23, 2022. Accessed: May 12, 2022. [Online]. Available:
https://en.wikipedia.org/w/index.php?title=Emanoil_C._Teodorescu&oldid=1084216765
[7] A. Ben-Amotz, J. E. W. Polle, and D. V. Subba Rao, Eds., The alga Dunaliella: biodiversity, physiology,
genomics and biotechnology. Enfield, NH: Science Publishers, 2009.
[8] S. Feng et al., “CRISPR/Cas technology promotes the various application of Dunaliella salina system,”
Appl. Microbiol. Biotechnol., vol. 104, no. 20, pp. 8621–8630, Oct. 2020, doi: 10.1007/s00253-020-
10892-6.

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[9] “Dunaliella Salina by Steve Gschmeissner/science Photo Library,” Pixels.
https://pixels.com/featured/1-dunaliella-salina-steve-gschmeissnerscience-photo-library.html
(accessed May 11, 2022).
[10] Anonymous 2022a, https://www.globenewswire.com/news-release/2019/10/15/1929461/0/en/Global-
Carotenoids-Market-is-expected-to-reach-USD-3-59-billion-by-2025-Fior-Markets.html, (accessed July 28,
2022)

[11] Anonymous 2022b, https://www.gminsights.com/industry-analysis/beta-carotene-market, (accessed

July 28, 2022)

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