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Investigating algae for CO2 capture and accumulation and

simultaneous production of biomass for biodiesel production

Abdul Hai Alami, Shamma Alasad, Mennatalah Ali, Maitha

Alshamsi

PII: S0048-9697(20)37060-1
DOI: https://doi.org/10.1016/j.scitotenv.2020.143529
Reference: STOTEN 143529

To appear in: Science of the Total Environment

Received date: 13 July 2020

Revised date: 21 October 2020
Accepted date: 27 October 2020

Please cite this article as: A.H. Alami, S. Alasad, M. Ali, et al., Investigating algae for
CO2 capture and accumulation and simultaneous production of biomass for biodiesel
production, Science of the Total Environment (2020), https://doi.org/10.1016/
j.scitotenv.2020.143529

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© 2020 Published by Elsevier.

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Investigating algae for CO2 capture and accumulation and simultaneous production of

biomass for biodiesel production

Abdul Hai Alami1, 2, *, Shamma Alasad1, Mennatalah Ali1 and Maitha Alshamsi1
1
Sustainable and Renewable Energy Engineering, University of Sharjah, P.O.Box 27272, Sharjah, United
Arab Emirates

2
Center for Advanced Materials Research, Research Institute of Science and Engineering (RISE),
University of Sharjah, Sharjah, P.O.Box 27272, United Arab Emirates

*corresponding author, email: aalalami@sharjah.ac.ae, phone: +971-56-160-5355, fax: +971-6-5585191

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Abstract

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Carbon capture and sequestration technologies are used to reduce carbon emissions. Membranes,
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solvents, and adsorbents are the three major methods of CO2 capture. One of the promising
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methods is the use of algae to absorb CO2 from flue gases and convert it into biomass. Algae

have great potential as renewable fuel sources and CO2 capture using photosynthesis for carbon
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fixation has also attracted much attention. This paper presents an extensive and in-depth report
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on the utilization of algae for carbon capture and accumulation. This is done in conjunction with
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cultivating the algae for the production of biomass for biodiesel production. Different systems

are investigated for algae cultivation as well as carbon capture to effectively mitigate carbon

emissions. The performance and productivity of these biosystems depend on various conditions

including algae type, light sources, nutrients, pH, temperature, and mass transfer. Macroalgae

and microalgae species were explored to determine their suitability for carbon capture and

sequestration, along with the production of biodiesel. The steps for producing biodiesel were

comprehensively reviewed, which are harvesting, dehydrating, oil extraction, oil refining, and
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transesterification. This technology combines active carbon capture with the potential of

biodiesel production.

Keywords: carbon dioxide capture; algae for CO2 capture; algae for biomass production;

biodiesel

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Table of Contents:

Abstract ...................................................................................................................... 1

1.Introduction............................................................................................................. 2

2.Methods of CO2 Capture…………………………………………………………………………………….4

2.1 Absorption……………………………………………………………………………………………..…...4

2.2 Adsorption…………………………………………………………………………………..…………………5

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2.3 Membrane Technology…………………………..……………………………………………………..6

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3. Using Algae for CO2 Capture………………………………………………………………………..……9

3.1 CO2 Capture using Photosynthesis……………………………………………………….……..9

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3.2 Algae cultivation Systems…………………………………...…………………………………..……………11

3.2.1 Open Systems……………….…..……………………………………………………………………..12

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Circular Ponds…………………………………………………………………………………………………………..13
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Raceway Ponds…………………………………………………………………………………………………………13

3.2.2 Closed Systems….……………………………………………….…………………………………..….14

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Tubular Photobioreactor…………………………………………………………………………………………..15
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Vertical Column Photobioreactor………………………………………………………………………….….16

Bubble Column……………………………………………………………………………………………….…18

Airlift…………………………………………………..…………………………………………...………………18

Flat Panel Photobioreactor……………………………………………………………………………………...19

3.3 CO2 Fixation Efficiency…………….………………………………………………………………21

3.3.1 Parameters affecting CO2 fixation efficiency…………………………………………22

Algae Types……………………………………………………………………………………………………………....22
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Macroalgae for Carbon Sequestration………………………………………………………..…….22

Microalgae for Carbon Sequestration……………………………………………………..…………23

Illumination Sources…………………………………………….…………………………………………...25

Nutrients……………………………………………………………………………………………………………………..27

pH……………………………………………………………………………………………………………………………….30

Temperature……………………………………………………………………………………………………..………...31

CO2 Concentration and Flow Rates……………………………………………………………………………...32

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Mass Transfer………………………………………………………………………………………………………..….…32

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3.4 Impact of Flue Gases on Algal Biomass…………………………………………………………….….....34

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CO2…………………………………………………………………………………………………………………….….…….35
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NOx………………………………………………………………………………………………………………….…….……..37
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SOx…………………………………………………………………………………………………………………..……………38

4. Biomass Measurements and Growth Rate……………………………………………………….39

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5. Harvesting and Biodiesel Production………………………………………………………………42

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5.1 Harvesting Techniques……………………………………………………………………………….42

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5.2 Drying and Dehydration……………………………………………………………………………..49

5.3 Oil
Extraction…………..…………………………………………………………………………………………..54

5.4 Oil Refining………………………………………………………………………………………………...58

5.5 Transesterification……………………………………………………………………………………..63

6.Conclusion…………………………………………………………………………………………………….….64

References……………………………………………………………………………………………………………66
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1. Introduction

Flue gases emitted from the combustion of fossil fuels are the main source of greenhouse gases,

as 68% of the total emissions are caused by carbon dioxide (CO2), contributing to global

warming and climate change [1]. Global warming is the increase in temperature in the lower

atmosphere that has already affected global climate and resulted in drastic changes in the glacier

mass at the poles, and hence the total reflectivity of the Earth, causing a vicious cycle with high

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volition [2]. To add to this, it was found that the increase in CO2 concentration as a result of

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human activity in industrialization and transportation is the leading cause of accelerating climate

change [3]. This changes manifests in phenomena that were not witnessed before, such as the
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rise in sea level and sporadic floods and droughts in adjacent locales [4]. Furthermore, the global
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concentration of carbon dioxide has increased from 280 ppm to 368 ppm in the past 200 years,
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which necessates a serious effort to develop novel technologies to reduce and mitigate emissions.

Carbon capture and storage (CCS) technologies have been developed for this purpose, and they
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devise effective ways of capturing CO2 being emitted from power plants and transportation
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sectors and storing it away for prolonged periods of time [5]. Depending on the location intended
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for capture, there are three approaches currently used for CO2 capture [3]:

 Post-combustion capture: where CO2 is removed after the combustion of a fuel.

 Pre-combustion capture: which involves separating and storing CO2 before combustion.

 Oxy-fuel combustion: where coal is burned in the presence of oxygen to produce a

concentrated stream of CO2.

These three methods are technologically complex, are expensive and are honed for industrial

settings [6]. More interest has grown in mimicking plant need for CO2 capture by the natural
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process of photosynthesis [2]. Biological post-combustion CO2 capture is also important since it

has the added benefit of benefiting from the produced biomass as a by-product [7]. From a

botanical viewpoint, more than 50% of total photosynthesis processes take place in algae, thus

algae represent a promising feedstock for CO2 mitigation with limited dependence on weather or

irrigation conditions. More interestingly, these macro and microalgae are also studied for

biofuels production specifically for of their ability to grow in diverse conditions [8] and

compared to terrestrial plants, many algal species are able to produce higher biomass yields [9].

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Researchers found that microalgae are able to sequester 10-50 times more CO2 than their

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terrestrial counterparts [2] be it from atmospheric air with low CO2 concentration or from flue

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gases with 12% to 15% of CO2 [10]. Some species are also able to tolerate high levels of SOx
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and NOx that are present in such flue gases [7]. Because of their high photosynthetic efficiencies,

algae are able to accumulate high lipid content that can further be extracted from the biomass to
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produce chemical energy such as biodiesel, which is a promising alternative to fossil fuels [11].
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Various authors have studied the combined benefit of using microorganisms for carbon dioxide
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capture and simultaneous biodiesel production [12]. For example, Klinthong et al. have

investigated the positive impact of using microalgae for CO2 capture, as it does not compete for
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land with other crops and at the same time its production is rapid with a good yield of biodiesel

[13]. In antoher study, Jalilian et al have studied the adaptation of macro- and micro- algae for

biofuel production and for mitigation of environmental pollution [8], indicating the sustainability

of the algae and their formidable contribution to CO2 bio-sequestration. Razzak et al also studied

the incorporation of microalgae cultivation with waste water treatment and CO2 capture [14]. The

benefits of such approach are numerous, including the contribution of such algae tor water

filtration, CO2 capture as part of their biological metabolic processes, and also their small
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footprint when it comes to land usage. Pavithra et al investigated the effect of microalgae not

only on biofuel production, but also as an added technique for removal of heavy metals from the

environment [15]. Mondol et al also reported on using metal organic frameworks (MOFs) to

enhance biofuel production and also the removal of nitrogen-containing compounds [16]. The

rate at which CO2 is being absorbed from by the algae is very important. Higher flow rates could

indicate less-than-effective absorption depending on the algae species, as indicated by the work

of Depra et al [17]. The growth kinetics of microalgae should also be considered and highlighted.

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Authors such as Aghaalipour et al [18] such parameters for specific algae strains at different

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carbon dioxide feeds, and observed that 10% CO2 feed yielded maximum growth kinetic

enhancements.
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This paper presents a focused discussion on technologies and setups that utilize algae as a
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potential medium for carbon dioxide capture. And since carbon dioxide is an important nutrient

for algae, then there is an added interest in methodologies to also cultivated algae and harvest it
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as useful biomass. This imposes a limit on the maximum operational temperature of carbon
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dioxide capture, but the added benefit of incubating the algae and producing biomass is an

extremely attractive route. In the following paragraphs, a brief outline on available carbon
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dioxide capture techniques is given, then a more focused analysis of algae-based capture

methods is discussed. Finally, methods to extract biomass from the algae are presented and

analyzed [19]. While the authors are aware that such approach have the promised advantages,

some disadvantages include the rapid growth of such algae that might not be too sustainable

along with the health and safety issues that they create. The biofuel itself will be combusted at

some future point, which in turn contributes to the emission issue that this technology is working

to eliminate.
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2. Methods of CO2 Capture

As discussed before, post-combustion is the most close-to-market CCS strategy [20] as shown in

Figure 1. Although many post-combustion CO2 capture technologies such as absorption,

adsorption, and membrane have been explored, none of them provides an ideal solution due to

the significant amount of flue gas emitted from power plants and mass transfer limitations in

these methods [21]. In addition, the storage of carbon dioxide in depleted oil and gas reservoirs,

mined salt caverns and deep ocean storage involve many problems such as the long-term

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integrity of the chosen volumes, and the risk of CO2 escaping back to the atmosphere.

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Furthermore, the costs of transporting CO2 to the storage sites is a major concern [2].

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Figure 1: Post-combustion capture [3]

2.1 Absorption
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Amine-based chemical absorption is the most developed CO2 capture method. This process starts

by supplying the flue gas to the solvent from the bottom for absorption, then the CO2-rich

solvent that exits is sent to the stripper for regeneration which releases pure CO2 when heated

that will be compressed for transportation and storage. Finally, the CO2-lean solvent, such as

Monoethanolamine (MEA) (solvent of choice for chemical absorption), is sent back to the

absorber to repeat the cycle [22]. After the CO2 is absorbed by the aqueous solution, the MEA

regenerates during desorption, where a reversible reaction takes place by high temperatures [23].

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The water vapor in the regenerated CO2 can be separated by condensation. The absorption and

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desorption reactions are written as [2]:

CO2 + 2C2H4OH ─NH2 C2H4OH ─NHC-p + C2H4OH─ NH3⁺ (1)

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Although this technology is one of the most mature (to date) for CO2 capture, there are several
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drawbacks, of which the high energy consumption for high temperature solvent regeneration

[21].
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Physical absorption where CO2 absorption process takes place without chemical reaction and
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based on Henry’s law, under high pressure and low temperature, then the desorption process
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occurs under low pressure and increased temperature. However, this method is less efficient due

to the low partial pressure of CO2 in the post combustion flue gas [21].

2.2 Adsorption

Another way that the capture CO2 could be accomplished is by using solid adsorbents

[24]. Carbon dioxide first diffuses into the pore of a mesoporous adsorbent in a rate higher than

that for amine absorption as the gas-liquid mass transfer is affected by the low contact area.

However, this technique has several disadvantages, including low adsorption capacities at low
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pressures and the impact of water vapor and other gases on the adsorption process. Many

materials are proposed for adsorption purposes, such as amine-based chemical adsorbents which

have been widely investigated. This is mainly due to their chemically modified active sites that

provide high surface area for favorable adsorption process [25]. The main drawbacks here are

high costs and the need of high CO2 adsorption capacity and are still the main challenges [21],

[25].

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2.3 Membrane technology

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Membranes offer good selectivity towards extracting specific compounds from a gas stream. For

example they show size-based selectivity of oxygen versus nitrogen in oxyfuel combustion
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systems or carbon dioxide from natural gas in natural gas processing, carbon dioxide from flue
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gas in post combustion system and carbon dioxide from hydrogen in pre-combustion system
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[26].

Membranes are considered to be semi-permeable materials that can be used to separate

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substances by different mechanisms such as ionic transport, adsorption/diffusion,

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solution/diffusion and molecular sieve [27]. The solution/diffusion process involves dissolving a

gas, sending it to the membrane surface and then spreading it by mass transfer across the

membrane. Molecular sieving introduces physical separation for separating smaller molecules

from the bigger ones by using a very delicate mesh mode [28]. Membrane perform as filters

which work on separating one or more gasses from a feed mixture and producing a gas-permeate

as it shown at Figure 2. Two factors are imposing the performance of the membrane which are

selectivity (The predilection of the membrane to allow for one gas strain to go over the other)
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and permeability (Which is the flux of a particular gas among the membrane) [26].Membranes

can be classified based on materials they are made of, which can be organic as polymeric or

inorganic involving metallic, zeolite, carbon or ceramic, also it may be porous or non-porous

[27].

Regarding the CO2 capture, membranes are categorized into two sorts: 1) Gas separation

membrane, and 2) Gas absorption membrane [27].

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Figure 2: Gas separation membrane schematic [26]

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Gas separation membrane

This type of membrane benefits from the preferential penetration the constituents of a gas

mixture across the porous structure of the membrane [29], which allows one of the component to

diffuse quicker than others through the membrane [27]. The prime design and operational factors

of membranes are permeability and selectivity [29] and for CO2 bearing combination, the CO2

contained in a gas stream is introduced at high pressure into a membrane separator that may
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consist of a large number of hollow cylindrical membranes ranked in parallel (see Figure 3). As

CO2 passes across the membrane is retracts on the shell side of the separator where the pressure

is reduced [27]. This design is simple and works for a steady state process and has no moving

parts. The membrane separation of CO2 from sprightly hydrocarbons is currently used in natural

gas, petroleum and chemical industries [30].

Gas separation membrane is not exclusively used for CO2 capturing from the flue gases because

of its need to flue gas pressurization and the relatively high mixture flow [29].

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Figure 3: Gas separation membrane principle [27]

Gas absorption membranes

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Gas absorption membranes are composed of microporous solid membranes as shown in Figure 4,

and they work on introducing the target gas to the absorbing liquid flow. It has been proven that

when CO2 separation is needed, it should first be diffused across a membrane and then uptaken

by a liquid absorbent. Due to the high driving force relating to large flow rates, this operation

leads to a higher removal rate than gas membrane separation [27]. The development of

independent monitors of liquid flows and gas flows has led to more efficient operation than the

usual separator membrane [29].

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Figure 4: Gas absorption membrane principle [27]

3. Using Algae for CO2 Capture

3.1 CO2 capture using photosynthesis

Algae sequester CO2 from the atmosphere during photosynthesis and recycle it in the form of

biomass [31]. Photosynthesis is a process that takes place in chloroplasts, by which solar energy

is converted into chemical energy in the form of lipids and carbohydrates, which can further be
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used for producing biofuels, food, or other chemicals [32]. The overall photosynthesis reaction is

written as [7]:

6H2O + 6CO2 + Photons (light) C6H12O6+ 6O2 (2)

The process is divided into two parts: a light-dependent reaction and light-independent one. In

the former reactions, solar radiation is converted into adenosine triphosphate (ATP) and

nicotinamide adenine dinucleotide phosphate (NADPH), both are molecules found in all living

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cells and organisms. ATP is an intercellular energy transfer compound and is consumed in living

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metabolic processes. On the other hand, NADPH mainly converts CO2 into glucose as it appears

in the last step of photosynthesis reactions [1]. The reaction occurs in the thylakoid membranes
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and consists of two photosystems that use excited chlorophyll molecule via sunlight energy to
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transport electrons [5]. The reaction starts in photosystem ІІ (PSІІ) through which water
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molecules are dissociated into oxygen ions, hydrogen ions, and electrons which are passed to

photosystem І (PSІ) [1], [5]. ATP and NADPH are formed by electrons and hydrogen
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transfer[5]. The electron transport chain provides energy for the synthesis of ATP, and NADPH
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is formed from NADP⁺ by the reduction of ferredoxin and the transfer of two electrons in PSІІ
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[33]. The overall light-dependent reaction is written as follows [13]:

2H2O + 2NADP⁺+ 3ADP + 3P + light 2NADPH + 2H⁺+ 3ATP + O2 (3)

The light-independent reaction (or dark reaction) begins with capturing CO2 molecules by

ribulose-1,5-bisphosphate (RuBP), catalyzed by ribulose-1,5-bisphosphate

carboxylase/oxygenase (Rubisco) enzyme, which determines the rate of carbon dioxide fixation.
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This step is also known as CO2 fixation. Then, the next step is the Calvin-Benson cycle. During

this step, organic compounds are produced as sugars from CO2 and H2O with 3 molecules of

ATP and 2 molecules of NADPH required to reduce each molecule of CO2 [34]. Finally, RuBP

is regenerated to continue the CO2 fixation process. The sugars produced can be converted

through metabolic reactions to amino acids and lipids [5], [33]. The overall light-independent

reaction is written as [13]:

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3CO2 + 9ATP + 6NADPH + 6H⁺ C3H6O3-phosphate+ 9ADP + 8P + 6NADP⁺+ 3H2O (4)

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When high concentration of dissolved Oxygen is produced as a by-product, Rubisco enzyme can
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use it for photorespiration. At this process O2 is consumed to produce CO2. In addition, the

generation of high concentration of O2 during photosynthesis can lead to algal growth inhibition,
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since the exposure to solar radiation, for example, can produce reactive oxygen species that may

damage cellular components and membranes [35].

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Carbon concentrating mechanism (CCM) increases the concentration of CO2 for Rubisco
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enzyme. When pure CO2 is dissolved in water, the pH drops due to the formation of carbonic

and bicarbonate acids, creating acidic media [5] . The equilibrium reactions are written as [11]:

CO2 + H2O H2CO3 HCO3⁻ + H⁺ C +2H⁺ (5)

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The Rubisco enzyme will only react with CO2, thus when it reaches saturation, another enzyme

called Carbonic anhydrase (CA) is used to convert the carbonic and bicarbonate acids into CO2,

and therefore into biomass [5]. This enzyme was found to be in both micro and macroalgal

species, allowing for active and passive carbon capture [32].

Furthermore, the carbon concentrating mechanism can significantly affect the growth and CO2

bio-fixation of algae. At high CO2 concentration, the activity of carbonic anhydrase (CA) can be

inhibited due to the increase in carbonic and bicarbonate ions and decrease in pH value of the

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medium. However, some algal cells are able to adapt by gene regulation and increasing the

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generation of adenosine triphosphate (ATP) to maintain pH stability inside the cell by active

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3.2 Algae cultivation systems

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Algae cultivation systems are classified into open systems, such as open ponds, where the culture

is directly exposed to the environment and closed systems, such as photobioreactors, where the
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culture is fully enclosed in a container. Table 1 summarizes the advantages and limitations of
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different algae cultivation systems.

Table 1: Advantages and limitations of different algae cultivation systems [36].

Cultivation Advantages Limitations

systems
Open ponds Relatively economical, easy to clean, good for mass Little to no control of culture conditions,
cultivation of algae culture easily contaminated, require large
land mass, limited to few strains of algae,
poor productivity
Tubular Relatively cheap, suitable for outdoor cultures, large pH gradients, dissolved O2 and CO2 along
photobioreactors illumination surface area, good biomass productivities the tubes, fouling, wall growth, requires
large land space
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Vertical column High mass transfer, high potential for scalability, low
photobioreactors energy consumption, low photoinhibition and photo-
oxidation, good mixing with low shear stress, good for
algae immobilization, easy to sterilize
Flat panel Good light path, good biomass productivities, low oxygen
photobioreactors build-up, large illumination surface area, good for algae
immobilization, easy to clean, suitable for outdoor
cultures, relatively cheap
Small illumination surface area,
illumination surface area decreases with
scale-up, shear stress to algae cultures,
require complex materials
Difficulty in scaling-up, difficulty in
controlling culture temperature, possibility
of hydrodynamic stress to some algae
strains, wall growth

Open Systems

Open systems are divided into natural waters (lakes and ponds) and artificial ponds. Mass

cultivation of microalgae in open systems has been used worldwide in the last few decades for

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biomass production [37]. It is commonly done in large ponds, circular ponds, and raceway ponds

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[38].

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Open systems are simple, cheaper, easier to build and operate than closed systems. Additionally,
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open systems are easy to scale-up which can be augmented by increasing the area not the depth .
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They have to be shallow to ensure microalgae receive sufficient light, since sunlight can only

penetrate limited water depth. Moreover, temperature and lightning cannot be controlled due to
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the system’s lack of operational controls. The major limitations of open systems include poor

light usage by the cells, low mass transfer rate due to inefficient stirring mechanisms, water
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evaporative loses, CO2 diffusion into the atmosphere, and the requirement of large areas of land.
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Bad weather can inhibit algae growth. In addition, open systems can be easily contaminated by

predators, alien species and microorganisms, which restrict algae production [7]. Thus, open

systems are only used for organisms that can tolerate extreme conditions, like high salinity or pH

[39]. Altogether, open systems have low biomass productivity of 0.05-0.1 g L-1 day-1 and low

cell concentrations of less than 1 g L-1 [40].

Circular Ponds
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Circular ponds are the oldest algae cultivation systems and are shown in Figure 5. However, they

are not preferred because they require expensive concrete construction as well as high energy

input for mixing using a rotating arm at the center of the pond.

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Figure 5: Circular ponds [41].
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Circular ponds operate at a water depth of 30-70 cm. The most commonly used algae for
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biomass production in circular ponds are Chlorella sp. [42]. The average biomass productivity

can reach about 15 g m-2day-1 in dry weight basis [43].

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Raceway Ponds

Raceway ponds are the most widely used algae cultivation systems. They are the most feasible

and cheapest to construct as they can be constructed using concrete, glass fiber, or plastic.

Raceway ponds consist of a closed loop with oval-shaped recirculation channels, where the flow

is directed around bends by baffles positioned in the flow channel, as seen in Figure 6.
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Figure 6: Raceway ponds [44].
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They are continuously stirred with a paddlewheel to provide a homogeneous culture and prevent

sedimentation [42]. Paddlewheels are the driving force of the flow. The culture is circulated
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around the racetrack, following the same direction as the paddlewheel. Moreover, the circulation
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rate can be adjusted by the paddle speed. Raceway ponds operate at a water depth of 15-30 cm.

Fresh feed, containing nutrients, is added in front of the paddlewheel. Algae broth is harvested

behind the paddlewheel after it had circulated through the loop [7]. The average biomass

productivity can reach up to 22 g m-2day-1 in dry weight basis [43].

Closed Systems

Closed systems or photobioreactors consist of an illuminated vessel or container that encloses the

culture and provide a controllable environment with no direct exchange of gases between the
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cultivation system and the outside environment [45]. Photobioreactors are flexible and versatile

with control systems installed for monitoring and controlling conditions like temperature, pH,

light intensity, nutrients, and CO2 concentration [46]. They are suitable for strains that cannot

tolerate extreme environment. Closed systems lower the risk of contamination, minimize CO2

losses, prevent water losses by evaporation, have higher biomass productivities and cell

concentration but at much higher costs [7]. Photobioreactors designed for CO2 sequestration

should have high mass transfer and the ability to use CO2 gas as a means of mixing and

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providing nutrients for algae growth [47]. In general, photobioreactors have high biomass

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productivity greater than 0.8-1.3 g L-1 day-1 and high cell concentrations greater than 1 g L-1 [40].

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photobioreactors used for microalgae cultivation, and those will be discussed in the following

sections.
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1. Tubular Photobioreactor
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Tubular photobioreactor is one of the most suitable configurations for outdoor mass

cultivation [48]. The tubes are usually made of transparent glass or plastic like acrylic,
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polypropylene, or polyvinylchloride that are long with small internal diameters to increase
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light penetration. An example is shown in Figure 7.

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Figure 7: Tubular photobioreactors [49].
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The diameters are normally 10-60 mm, the length can extend to several hundred meters, and the
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liquid velocities are 0.2-0.5 m/s. Tubular photobioreactors can be arranged in different forms and

orientations including vertical, horizontal, helical, and inclined, which helps in achieving high
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light conversion efficiency [50]. Horizontal tubular photobioreactors have a larger surface area to
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volume ratio and a better incident light angle than vertical. The tubular system is comprised of

several components including solar array for algae growth, harvesting unit for separating algae

from the suspension, degassing column for gas exchange, and circulation pump for culture

mixing [35]. However, photolimitation can occur in tubes with large diameters because cells in

the center receive less light, which restricts their growth. Furthermore, mass transfer problems

occur in long tubes, resulting in high concentrations of dissolved carbon dioxide at the aeration

zone and high concentrations of dissolved oxygen at the degasser zone. In summer, high

temperature will increase in the culture due to the limited volume from the small diameters,
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therefore making it difficult to scale-up [50]. Other disadvantages of tubular photobioreactors

include high energy consumption, oxygen accumulation which can inhibit photosynthesis, CO2

depletion, pH gradient, fouling, the possibility of microalgae growth on the tube walls, hence

blocking light, and the requirement of large land space as the tubes spread over a large distance.

Nevertheless, they have large illumination surface area, hence a good biomass productivity. As

well as a better air-residence time inside the photobioreactor, which can provide more dissolved

CO2 [7].

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2. Vertical Column Photobioreactor

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Vertical column photobioreactors are transparent vertical tubes made of glass or acrylic. They
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are rigid and have bigger diameters of 15-20 cm, so a gas sparging system is needed. Bubble

column and airlift are two types of vertical column photobioreactors that have sparger at the
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bottom of the column as seen in Figure 8. A sparger is used to convert inlet gas into tiny bubbles
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that will rise upwards, creating turbulence that provides the required mixing and mass transfer of

CO2, as well as the removal of oxygen produced during photosynthesis [7].

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Figure 8: Vertical column photobioreactors [51].

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This type of mixing exerts low shear stress and has low energy consumption. Vertical columns
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are compact, easy to clean and maintain but have a smaller surface area of irradiance. Internal

light sources can be installed inside the column; however, it is not cost effective. Furthermore,

this type of photobioreactor is expensive because of the materials used in the construction of the

column in order to make it sturdier. Nevertheless, it has high mass transfer and biomass yield

[52]. Vertical column photobioreactors have no physical agitation system implemented in its

design. Hence, very little cell damage is associated with this photobioreactor. Tall columns can

form CO2 gradients, which deprive algae of CO2, thus creating pH gradients. Moreover,
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increasing the length increases the residence time of oxygen that was produced by

photosynthesis, reaching to a level that is inhibitory [35]. Diameters of 20 cm and more leads to

high dark fraction in the middle of the column. This zone does not contribute to productivity and

might have damaging effects on growth [53].

3. Bubble Column

Bubble column is the most common type of vertical column photobioreactor with a height more

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than twice its diameter. It has a high surface area to volume ratio, a good heat and mass transfer,

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a homogeneous culture environment, and an efficient oxygen release. In addition, it has a low

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capital cost. The gas bubbles rising upward from the sparger provide the required mixing and gas
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transfer. Depending on the design of the column, perforated plates may be installed to break up

bubbles and increase turbulence. Outside light is often used as the source of illumination.
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Nonetheless, an inner illumination can be used for a higher light penetration efficiency and
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uniform light distribution. Photosynthetic efficiency depends on the gas flow rate and the light

and dark cycle that form when the liquid is circulated from the central dark zone to the external
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light zone at a high gas flow rate. This circulation flow pattern does not happen when the gas
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flow rate is less than 60 m/s because of the lack of back mixing. Increasing the gas flow rate

results in a shorter light and dark cycles, thus increases the photosynthetic efficiency [7].

4. Airlift

Airlift is another type of vertical column photobioreactor that consists of a vessel with two

interconnecting regions: riser and downcomer. The gas riser tube is where the gas mixture flows
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upward from the sparger to the surface, while the downcomer does not receive the gas but the

medium flows toward the bottom and circulates within the riser and downcomer [7].

Internal loop and external loop are two forms of airlift photobioreactor. The internal loop

photobioreactor can be further classified into internal-loop split airlift reactor and internal-loop

concentric tube reactor. The regions in an internal loop reactor are separated either by a draft

tube or a split cylinder. Whereas in an external loop, the regions are separated by two different

tubes. The sparger in the riser tube mixes the medium by bubbling the gas, causing it to move

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haphazardly upwards. This enables the liquid to move upward due to the decrease in the riser’s

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density [7].

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Airlift creates a circulation flow where the liquid culture flows continuously through light and
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dark phases, giving a flashing-light effect to the microalgae cells. The biomass productivity

depends on the light zones and operating time. Moreover, the performance is controlled by the
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residence time of gas in the different zones [7]. Airlift photobioreactor has the advantage of
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excellent mixing, which allows cells exposure to light radiation despite having large diameter

column and high cell density. Therefore, airlift has a better biomass production of microalgae
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than other vertical column photobioreactors [35].

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5. Flat Panel Photobioreactor

Flat panel photobioreactors are thin rectangular reactors made of transparent materials such as

glass, plexiglass, polycarbonate, and optical light film. They are characterized by large

illumination surface area and short light path. Therefore, they have low oxygen accumulation.

Furthermore, flat panels have a high surface area to volume ratio, as well as an open gas

disengagement system [7] as shown in Figure 9.

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Figure 9: Flat panel photobioreactors [49].

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There are various dimensions of flat panel, but height of lower than 1.5 m and width less than 0.1
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m are preferred. In addition, the panel’s depth should be below 0.07 m in order to maintain
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average irradiances above 100 E m-2 s-1 and to maintain cell density of about 1 g l-1 [54]. Flat

panel photobioreactors can be categorized into pump driven and airlift. Pump driven depends on

the liquid flow created by pumping to generate turbulence for mixing. Whereas airlift depends on

compressed air to provide the power of mixing. The thickness of the panel is an important

parameter in the design of the photobioreactor because it determines the surface area to volume

ratio and the length of the light path. The smaller the thickness, the better the distribution of

light. Moreover, a short light path or thickness allows for a higher optimal cell density and
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biomass productivity. However, thin photobioreactors are expensive, hard to clean, and subjected

to light inhibition and temperature fluctuation. The tilt angle is another parameter to consider in

flat panel photobioreactors. The optimal tilt angle of the reactor that allows maximum incident

light will change throughout the year due to the position of the sun [35]. Specifying the optimal

position of flat panels, particularly the direction, tilt angle, and distance between the panels,

significantly determines the overall system yield [54]. Mixing can be performed by using a

perforated tube to bubble air or by using a motor to mechanically rotate the panel. Flat panel

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photobioreactors have high biomass productivities and high photosynthetic efficiency. However,

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it is hard to control the temperature of the culture and scale up because increasing the volume

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will increase the hydrostatic pressure. And the system cannot withstand high pressure. Flat
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panels can be easily combined together but it might face problems with relatively high space and

light energy requirements, thus, the possibility of a low mass production per unit of space
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efficiency [7].
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3.3 CO2 Fixation Efficiency

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Pure CO2 is not required for algal growth cultures. CO2 can be sequestered by algae from a
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variety of flue gases emitted from combustion processes [55]. Carbon dioxide capture efficiency

depend mainly on the type of strain selected, the concentration of CO2, cultivation system,

environmental and operating conditions. Biomass and growth rates measurements are essential

for evaluating the bio-fixation efficiency [56]. Under optimal conditions, CO2 capture

efficiencies can reach as high as 80% to 99% [57]. Furthermore, at high CO2 concentrations,

algae can enhance their adaption when the supply is gradually increasing. Therefore, slowly

increasing the CO2 supply will cause a higher fixation and growth rate [7]. Usually, different
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CO2 concentrations are used for determining the CO2 fixation efficiency. The removal efficiency

is determined by:

(6)

The rate of CO2 fixation can be determined from the amount of carbon in algal biomass. It can

be estimated from:

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RCO2 (g CO2 ) = Cc (7)

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Where is the volumetric growth rate in linear growth phase (g dry weight ), MCO2
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and MC are the molar mass of CO2 and elemental carbon, respectively, CC is the average carbon

content of the biomass (% w/w). For microalgae measured by an elemental analyzer, obtained
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0.507 g carbon per g dry cell weight [56], and another simplified method assumed that each 1 kg
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of produced biomass equals 1.88 kg of recycled CO2 [58] .

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3.3.1 Parameter affecting CO2 fixation efficiency:

Algae Types

Algae are categorized into two types: microalgae and macroalgae (seaweed), depending on their

size [59]. They could be found in marine water, fresh water, or rocky shores. Chemical or

physical parameters, such as light, temperature, carbon dioxide, pH, aeration, and salinity could

affect algae growth [8]. The process of combining carbon sequestration with biomass production
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could be used for faster removal of CO2 from the atmosphere [60]. Therefore, both types were

employed for carbon dioxide sequestration and biofuel production [61].

Macroalgae for Carbon Sequestration

Macroalgae, known as seaweed, is a multicellular algae that grows largely in marine

environments [59] . Marine macroalgae have high calorific value which makes them an

alternative for the production of fuels and other chemicals [62] . They have greater growth rates

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than terrestrial plants, but lesser than microalgae. In addition, they are rich in carbohydrates and

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better used for producing biofuels like bioethanol and biogas. However, since macroalgae can be

used for carbon capture along with the accumulation of biomass, it would be interesting to
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develop genetic tools to move its metabolic pathways towards the enhancement of lipid
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production, which can further be used for producing biodiesel [63].
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The kelps, Macrocystis and Laminaria, are types of marine macroalgae that have a very high
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photosynthetic rates and productivities, therefore, they have the potential to make a significant

contribution towards CO2 capture and storage [64]. Also, it was found that species of Sargassum,
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Ascophyllum, and Fucus have the highest photosynthetic rates in the phaeophytes algal group,
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while Porphyra and Palmaria have faster photosynthetic rates in the rhodophytes. From the

chlorophytes, Ulva and Enteromorpha can also achieve high rates of CO2 capture per gram fresh

weight [9]. The first study done on freshwater macroalgae, Oedogonium, to identify the effect of

CO2 on the biomass production and carbon capture in large-scale outdoor tanks, found that the

low efficiencies of carbon capture when CO2 is delivered on-demand were higher than

microalgal efficiencies when flue gas is continuously added. In addition, by supplying a higher

concentration of CO2, both the biomass productivity and the amount of carbon sequestered per

unit area increased, but the efficiency of carbon capture decreased. However, without the
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addition of CO2, the biomass fixed carbon at a rate of 481 g C , while when supplied

with CO2 at a pH 7.5, the rate of carbon fixation was 1.38 kg C . This specie has a

carbon content of 41-45% which is higher than most of marine seaweeds (22-35% Carbon),

making it a candidate feedstock for carbon capture and conversion into biomass [65].

Microalgae for Carbon Sequestration

Microalgae are unicellular microorganisms that have a variety of applications in different

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industries[8]. Unlike macroalgae that have low lipid content, microalgae can accumulate high

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lipid content, which makes it a target feedstock for the production of biodiesel [8], [60].

Microalgae have high photosynthetic efficiencies, which means more CO2 consumption, and
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high growth rates. They are capable of doubling their biomass every 4-6 hours, and can
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withstand various environmental conditions. Depending on the specie, the oil content can reach
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80%. Therefore, microalgae can be used for CO2 storage and conversion to fuels [60] .

Microalgae are able to survive under a variety of carbon-rich conditions, which is reflected in the
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diversity of cellular lipids obtained from them. Researchers investigated the impact of high CO2
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levels on different species, the desired species were the ones that tolerate high concentrations
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coupled with high biomass production such as, lipids or triglycerides [11]. The common species

used for carbon capture include freshwater microalgae, such as Scenedesmus, Spirulina platensis,

and Chlorella. Recent research done on marine microalgae, mainly concentrate on several

species, such as Nannochloropsis, Dunaliella salina, and Isochrysis galbana [66], [67].

Marine microalgae are capable of surviving under extreme conditions, such as high salinity

environments and high temperatures, and generating higher amounts of crude oil than other

plants [68]. A research was done on two species with high biomass and oil content, which are

Isochrysis galbana and Nannochloropsis, with lipid content of 7.0%–40.0% and 22.7%–52.0%,
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respectively, where the two species were able to grow under different CO2 concentrations from

flue gases [69]. The maximum biomass concentration of the two species was under the 10% CO2

concentration, with (0.71±0.08) g/L and (0.78±0.03) g/L [66]. However, different cultivation

conditions affect the metabolism and CO2 fixation of the species. In another experiment done on

Nannochloropsis, it was observed that the biomass was able to grow better when supplied with

soluble sodium carbonate/bicarbonate derived from flue gas than when using only dissolved CO2

gas as carbon source. The dry biomass maximum yield was 0.55 g using the 20% carbonates

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solution, while the maximum biomass weight obtained using 15% dissolved CO2 was around

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0.44 g. This indicates that CO2 capture from industries using alkali absorption and storing it in

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liquids as carbonates and bicarbonates is an alternative way to enhance the growth of microalgae
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[11].
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Fresh water microalgae, such as Chlorella sp. was able to grow under a CO2 concentration up to

40%, whereas the highest biomass concentration of 2.05 g/L, was achieved at a concentration of
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10% v/v [5]. Scenedesmus sp., on the other hand, was able to survive under 100% CO2 and the
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biomass concentration has increased to 3.65 g/L in 30 days, while it was only 1.19 g/L using

atmospheric CO2 (0.036 %) [1]. The growth rate and CO2 fixation rate of Spirulina sp. were
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found inversely proportional to the concentration of CO2. High CO2 concentration may enhance

the mass transfer, however, the decrease in pH may inhibit the algal growth [57].

Illumination sources

The rate of growing of algae is controlled by a group of environmental factors in the culture

system as temperature, photoperiod, nutrient and light [70]. Light is one of the basic energy
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sources that algae need. As usual, cells obtain and store the light energy which affects the

capacity of the carbon fixation, and depending on it, can determine the rate of the cell growth

and its productivity. Light has impact on algae productivity as well as on its biomechanical

profile [7]. For enlighten the culture regime artificial light, sunlight or both of them can be used

[71]. Since light sources have a different spectrum from the sunlight, indoor validate models

probably are not viable to the outdoor cultures [72]. Sunlight is used in either closed or open

systems. While artificial light is used mainly in closed systems [7]. A study has investigated the

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effect of light sources as LED and fluorescent computationally and experimentally through

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modeling on the algae growth, to see how it affects the growth rate. Growth on fluorescent

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source has been found to be significantly higher than using other sources[73]. A review article
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mentioned primary factors that help in determining the microalgae cultivation rate of growth,

which are photoperiod (dark and light cycles) and light intensity [70]. Light and microalgae
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growth show a convoluted relationship and this relation could be presented in two modes: (i)
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light intensity and (ii) light-dark period cycle.Three stages are categorized in the effect of light

intensity: light inhibition, light limitations which are also called compensated light intensity, and
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light saturation [7]. Lower than this stage, light is considered as a limiting factor for algae
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productivity. At this point, the maximum rate of photosynthesis is obtained and rate of algae

growth at the light saturation stage is stabilized [74]. For determining the photosynthesis

efficiency and utilization efficiency, intensity of saturation light can be used, which is considered

as an important factor. Range of light intensities are generally used, about 100 to 210 μE/m2/s,

and saturation light intensity ranges is around 140 to 210 μE/m2/s. There are types of algae with

phycobilisomes that require low illumination, while other types require high illumination. For

example: Chlorella sp. and Scendesmus sp. have a saturation light intensity about 200 μE/m2/s
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[7]. Another study chose a certain algae species to investigate the effect of light intensity on their

growth. The results of the study revealed that light irradiance ranged from 33mmol to 400

mmol . For Selenastrum minutum, the growth rate was reported to be 1.73 at an

irradiance of 420 mmol and is considered as the maximum range. Minimum rate of growth

was found at irradiance of 200 mmol for Botryococus braunii [73]. For flasks, light

intensity is 1000 lux and around 5,000 to 10,000 for large volumes (see Figure 10). For

regulating light intensity, should be correlated with species density. When having low cell

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density, high light intensity will cause photoinhibition, while for high density species will have

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limited light penetration [71]. To sum up, cells density should increase in order to increase

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intensity of light provided [7]. For maximizing photosynthesis in fluorescent tubes, it is
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favorable to use blue and red spectrum [71]. Light is one of the important sources for

photosynthesis activity and autotrophic growing. Algae species that contain chlorophyll a and b,
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which are considered as the main light harvesting pigments, are affected by red and blue light.
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Some studies showed that green algae grow better than brown and red algae, since it composes

of chlorophyll a and b [74]. In terms of exposure time, it is determined that eighteen hours per
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day is the minimum range for artificial enlighten [71].

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Figure 10: Effect of light intensity on algae growth [7].
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Depending on engineering studies, it showed that the reactor geometry can decrease the
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microalgae suspension to light. Some studies showed the plane and circular geometry

implication, pointing that circular geometry is better in allowing for light infiltrate comparing to
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plane geometry. Thus, it will receive enough amount of light through allowing volume fraction
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of the reactor. Even though, in outdoor situation, plane geometry provides more uniform

distribution of light. Light afforded can be considered as hegemonic agent for identifying

productivity of photosynthesis. Another factor can be taken in consideration is culture

temperature. It can impact the algal cell’s metabolic reaction, thus needed light intensity can be

adjusted. Microalgae can grow normally under low light intensity demands in case of higher

temperature culture [7].

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Some researchers studied the growth of Nannochloropsis species under different light intensities

and wavelengths. Primary monochromatic light wavelengths are green, blue, white and red

LEDs. The ascending order for growing algae for LEDs was blue, white, green, red. The species

of Nannocloropsis reached the highest rate of growth at 0.64 and 0.66 in phototrophic

and mixotrophic culture respectively, beneath blue light [74].

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Nutrients

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Productivity of ecosystem is generally determined by nutrients and light entering the system.

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Some studies show that the elemental ratios in terms of qualitative output can be specified by

balancing nutrients and light [75]. Main nutrients for algae are phosphorus and nitrogen to
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preserve algae growth [71]. Other nutrients are cyanocobalamin, biotin, vitamins and various
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trade metals [7].

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In Oklahoma lakes and rivers, the elements found in water and considered as variables for the

growing of algae were nitrogen, phosphorous, calcium, potassium and magnesium. In this study,
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the nutrient elements were represented into two levels, high and low. It demonstrated that the
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growing of algae was low at low concentration of nitrogen and wasn’t effective as at high levels

of nitrogen. While at both levels of phosphorus, the effects were very similar [76]. Production

demands for CO2, land and nutrients depend on the mass balance evaluation [77].

Phosphorus and nitrogen can be found in both air and water. They are indigenous part of marine

ecosystem [71]. As an assumption, algae biomass typically composes of (C106H181O45N16P),

that means a 16 N:P (7.3 g N:1 g P) of fertilizer will be needed. Depending on the existence of

nutrients in the cultivation system and algae species, the N:P ratio can be varied from around 4:1
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to nearly 40:1. However, at low ratio of N:P in the high rate ponds of wastewater treatment, the

high productivity can be obtained [78]. A study revealed that the highest lipid production

achieved when the algae intake the nitrogen totally from wastewater. However, it declined

greatly after additional nitrate was added [79].

Nitrogen has a direct connection to the basic microalgae metabolism [7]. Nitrogen forms in the

shape of nitrite, nitrate and ammonia salt or ammonium [71]. Ammonium is favorable more than

nitrates due to the quick growing of algae and considered as primary source of nitrogen.

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Enhancing growth of microalgae can be done by supplying the medium by nitrate, if it was

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lacking nitrates. In case of nitrogen deprivation, microalgae will be able to produce more lipids,

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but growing in lower rates [7]. Nitrogen is a significant parameter in regulating the lipid content
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of algal cells[78]. The lipids are compounds that can accumulate in difficult conditions, even if
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the productivity is low [7].

Phosphorus can be provided at the form of dihydric phosphate, phosphate and hydrogen
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phosphate [71]. Combining phosphor with metal ions can be unsuitable for algae. Highly low
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concentration of P and N leads to algae growth inhibition. On the other hand, high concentration
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results in decreasing the rate of the growth, and in worst case killing the algae. Generally,

inorganic and chemical fertilizers provide nutrients. Benefit of using chemical fertilizer is

helping in lowering the pollution rates in the cultivation medium, leading to foster water reuse

for re-cultivation of algae [7]. Several researches suggest growing algae in wastewater to take off

nitrogen and phosphorus, as water consist of large quantities of phosphate and nitrate.

Recent US wastewater and fertilizer resource availability gathered with results at baseline

modeling representing the requirements of P and N which shows a fender to the development of

microalgae large scale based biofuel. Results of baseline indicate that wastewater sources can
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offer enough nutrients to generate around 3.8 billion gallons of fuel yearly, which is

approximately equal to about 6 percent of the DOE target of 60 billion gallon annually [77].

pH

pH control is important in obtaining high biomass productivity. pH greatly affects the growth of

algae. Algae species have different optimal pH ranges under which they grow. The optimal pH

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of most algae species is between 7-9 [35]. pH is a function of CO2 inputs, as it changes according

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to the concentration of CO2 within the culture. And an appropriate pH level is necessary to

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obtain maximum lipid content from the algae. The pH level can be controlled by increasing or

decreasing the CO2 concentrations. Furthermore, the more algae there is, the less carbon dioxide
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remains in the culture [52]. In the cultivation of a high-density microalgae with aeration from a
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pure CO2 or from flue gases with a high CO2 content, the CO2 dissolves in the culture,
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decreasing the pH and turning it into an acidic medium. Then, the CO2 is absorbed by the

microalgae during photosynthesis, thus increasing the pH of the culture [50]. There is no notable
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change in pH from atmospheric CO2 [7]. However, the transfer of NOx, SOx, and CO2 from the
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flue gas into the algae culture can cause pH to drop significantly if not properly controlled [80].

Combustion flue gas causes the pH to drop to about 5.5 with a CO2 concentration of 14% or

more. In addition, the presence of 100-250 ppm SO2 in the culture reduces the pH to about 3.5-

2.5 at an aeration rate of 0.25 vvm. SOx have a strong influence on algae growth. So, an extreme

pH change will restrain algae growth. Buffer solutions and active pH control can help prevent pH

reductions [7]. Sodium bicarbonate is used as a buffering agent to control the culture’s pH.

Hydrochloric acid and acetic acid can help keep the pH from rising excessively beyond the

tolerance of the algae [81]. Moreover, NaOH can be added to the nutrients to maintain a neutral
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pH, which is cost effective [80]. CO2 capture’s efficiency and productivity depends on the pH of

the culture [55].

Temperature

Temperature has a great impact on the system’s performance. It depends on the

environmental conditions and cannot be adjusted if the photobioreactor was placed outside. If

inside, it would be maintained at room temperature, particularly between 20-25 ºC [52].

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Temperature simultaneously impacts three competing cellular processes of algae, namely

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photosynthesis, photorespiration, and endogenous metabolism [35]. Temperature significantly

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affects speed of growth and reproduction. High temperatures usually accelerate metabolic rates
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of microalgae, while low temperatures can cause inhibition of microalgae growth. The optimal
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growth temperature of most microalgae species is in the range of 20-30 °C. When the

temperature is much higher or lower than the optimum, the specific growth rate of microalgae is
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decreased [5]. Many microalgae species are able to tolerate temperatures up to 15 °C lower than
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their optimal, however, temperatures lower than 16 °C will slow microalgae growth. On the other

hand, exceeding the optimum temperature by only 2-4 °C can result in total culture loss. There
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are many species that cannot tolerate temperatures above 35 °C because it is usually lethal to

them, whereas, some species can tolerate up to 60 °C. For example, Chlorella KR-1 and ZY-1

can grow at temperatures up to 40 C, and Chlorella sorokiniana UTEX 1230 can grow at 42 C

[7]. Developing a reliable and cost-effective temperature control mechanism is essential in a

photobioreactor design. Without temperature control, the temperature in a photobioreactor would

reach 10-30 C above the ambient temperature. Therefore, cooling mechanisms are often utilized

to keep the culture within a certain range. Some of the cooling methods include submerging the
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culture in a water pool, spraying water, shading, or incorporating a heat exchanger with the

photobioreactor [35].

CO2 Concentration and Flow Rates

Algae can tolerate CO2 to a certain level before it starts to limit its growth. This is due to two

reasons: 1) High CO2 concentrations can cause environmental stress that affects the capacity of

algal biomass to capture CO2, and 2) reduction in pH due to the formation of bicarbonate acid

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[7].

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At high flow rates, more CO2 is supplied to the algae. An experiment was conducted using
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different flow rates of pure CO2 added to the cultures of Desmodesmus sp. microalgae. It was
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observed that at a very high flow rates, the growth rates and dry biomass weight reduced after 12
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days. At 250 ml/min flow rate, the maximum dry weight was 70.01 mg/l with a specific growth

rate of 0.28 per day. The optimum flow rate was 50 ml/min, which achieved the highest dry
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weight of 496.81 mg/l and growth rate of 1.26 per day. At the lowest flow rate used, which was
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10ml/min, the dry weight was 357.75mg/l and a growth rate of 0.61 per day [2].
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In another study done on Chlorella sp. microalgae, at 1.75% CO2 air ratio, the maximum specific

growth rate occurred at a flow rate of 50ml/min with 1.11 , and the best CO2 fixation was

44.7 after 16 days. On the other hand, using 30ml/min flow rate resulted in a 100% CO2

fixation, but the growth rate efficiency was not high due to the lack of CO2 [10].

Mass Transfer
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Carbon dioxide mass transfer is related to algae growth and carbon dioxide reduction

effectiveness. Carbon dioxide mass transfer coefficient or kLa (CO2) presents mass transfer

condition occurring in the reactor. kLa (CO2) is a hydrodynamic parameter often used to evaluate

photobioreactor performance in microalgae cultivation process. This process demands an optimal

kLa value, where a high kLa indicates a better CO2 mass transfer within the culture. CO2 transfer

efficiency affects CO2 biofixation to improve microalgae productivity in the system [5]. The

photosynthetic production of algae is always followed by the production of oxygen and the

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consumption of carbon dioxide. Oxygen level above air saturation can inhibit photosynthesis in

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many algae species. Moreover, high levels of oxygen and irradiance can result in severe photo-

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oxidation which reduces the yield of the culture. So, it is important to establish good mass
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transfer capacity and photosynthesis rate [54].

The mass transfer coefficient, kLa, is the rate at which the gases, particularly air and carbon
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dioxide, is diffused into the medium.

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kLa = ( ) (8)
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Where kLa is the mass transfer coefficient (s-1) and PG/VL is the power input (W/m3) [52]. The
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term kLa is generally used to describe the overall volumetric mass transfer coefficient in

photobioreactors. It depends on several factors such as agitation rate, type of sparger, superficial

gas velocity, temperature, and surfactants [36].

The mass transfer from gas to liquid phase is given by the following equation:

(9)

Where is mass transfer rate, kLa is mass transfer coefficient, C* is equilibrium gas

concentration at the gas-liquid interface, and C is gas concentration in the solution [82].
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Mass transfer is a complex process in microalgae culture that involves three phase mass transfer,

which are: Gas (CO2) to liquid (culture medium); gas (CO2) to solid (microalgae); and liquid

(culture medium) to solid (microalgae). The mass transfer from gas to liquid phase is the major

limiting step in microalgae cultivation, since CO2 has a low mass transfer coefficient. The

oxygen produced during photosynthesis can inhibit microalgae growth. A typical solution is to

provide the gas with high flow rates to work in turbulence [7]. Sparger is installed at the bottom

of the photobioreactor to convert gas with different CO2 flow rate into small bubbles in order to

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improve mass transfer [5]. Mass transfer performance and biochemical reaction rate depend on

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CO2 concentration, bubble size, gas holdup, gas-liquid contact area, and gas-liquid ratio. Mass

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transfer can be increased by enhancing CO2 concentration gradient using NaHCO3 to generate a
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chemical reaction and by expanding contact area using hollow fiber membranes [7].
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3.3 Impact of Flue Gases on Algal Biomass

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Flue gases are considered as worthy carbon sources for algae photosynthesis, but they’re also
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composed of other chemical components involving heavy metals. In addition to CO2, flue gases

contain around 142 various compounds [83]. Knowing the flue gas composition is necessary and
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must be controlled due to its variety depending on the combustion system, generation source and

temperature of the combustion. Flue gas composes of some toxic compounds as SO2 to the algae.

In comparison, it contains of certain compounds as NO which can be metabolized.

The benefits of using CO2 from the flue gas, instead of that diffusing from the atmosphere is

returning to the insufficient amount of CO2 in the atmosphere that is not able to achieve the high

productivity in the biomass [84] and it is limited by the low concentration of CO2 in the air

which is around 0.03 to 0.06% CO2 [83].

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CO2

Effective capturing for CO2 from the flue gas at the power station restricted by the

algae’s tolerance to elevated CO2 levels. Generally, growth of microalgae needs around 10

volume percent of CO2 concentrations. For some species, more CO2 concentration tends to be

detrimental and lead to reduce the growth rate of biomass. Several studies had indicated that high

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CO2 levels inhibit the production of algal biomass and contribute in decreasing the efficiency of

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photosystem II. (Photosystem II: It’s a functional unit used in capturing photons from the light

energy to remove the electrons from water molecules).

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However, it is known that high levels of CO2 is desirable, although some species under high
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levels of CO2 do not exhibit substantial increase in the productivity of biomass. While others
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species display high tolerance for CO2 concentrations which can reach 100 vol% CO2 [84].
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When a study tested an increase in CO2 supply by 15%, it was found that Nannochloropsis sp.

showed a high production of biomass and substantial increase in its growth rate after about 2 to 4
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days. Another study found a high increase compared to Chlorella pyrenoidosa when CO2
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concentration was increased up to 20 percent [85]. Providing cultures with CO2 in various forms

as compressed CO2 gas, carbon, salts or bicarbonate help in improving the production of the

biomass by 260 percent and production of photosynthesis by around 256 percent, also it

enhances the lipid production. Microalgae absorb ranges around 20% to 70% of CO2, which

requires about 1.3 to 2.4 Kg CO2 per Kg of dry microalgae [83].

Some of the suggestions for improving CO2 tolerance in microalgae were by making genetic

modification through carbonic anhydrase or by adding bases as nitrates to prevent CO2

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acidification. Having low flow rates with high flue gas concentrations will result in a lower

concentration of dissolved inorganic carbon in the cultivation medium. Inorganic carbon source

contribute in enhancing the carbohydrate, lipid and biomass productivity [85]. Limitations of

mass transfer while sparging CO2 to the culture system, CO2 absorption kinetic and the need for

constructing the system of cultivation near a power station to avoid the cost of transporting CO2

are considered as barriers to algae CO2 fixation using flue gas [84]. However, bubbling CO2 with

flue gases offer an alternative for reducing the cost and at the same time contribute in reducing

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the greenhouse gas emissions [83].

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The rate of CO2 supplied in flue gases ( can be defined as [86]:

-p (10)
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), ρCO2 is density of CO2 (
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where is flue gas volumetric flow rate ( ), is

CO2content in flue gas before crossing the suspension (vol.%). CO2 assimilated in the
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suspension had been partially used by photosynthesizing algal cells and partially lost owing to its

escape into the atmosphere. The rate of supplying CO2 related to a unit of culture area A can be
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expressed by [86]:
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(11)

is the consumption rate of carbon dioxide consumed by algae (g ) within a unit

of culture area, is the liquid phase mass transfer coefficient ( ) for CO2 transfer from

the suspension into the atmosphere, is Henry’s constant for CO2 ( ), is

the mean partial pressure of CO2 in algal suspension (kPa), is the partial pressure of CO2 in

ambient atmosphere (kPa), Dec is degree of flue gas decarbonization after crossing the gas
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stream suspension (%). The mass transfer coefficient for CO2 was determined from the equation

[86]:

(12)

( ) is the mass transfer coefficient for transferring the dissolved oxygen from the

suspension to the atmosphere. and are the diffusion coefficients for CO2 and O2 in the

suspension ( ). The partial pressure of dissolved carbon dioxide is determined to be

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reduced exponentially from the beginning ( ) to the end ( ) of the cultivation area. Thereby,

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the formula utilized in calculating the mean pCO2 in the suspension of cultivation area is shown

below [86]: -p
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(13)
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Utilization (%) by the algae from CO2 supplied with flue gas can be expressed as [86]:
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(14)
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NOx

Nitrogen oxide and nitric oxide are the two main forms of NOx. They typically exist in the flue

gases. NO concentrations range between 90 and 95 vol% and NO2 is around 5-10 vol%. One of

the most significant nutrients for the growth of the biomass besides the carbon is Nitrogen.

Nitrogen can be consumed in different forms as N2, , , NO and [84]. One of

the parameters usually used for increasing the content of the carbohydrate in algal cells and the

lipid content is nitrogen deprivation [83]. For example Scenedesmus obliquus, Chlorella vulgaris
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and Chlamydomonas sp. demonstrated high lipid content and productivity under the conditions

of exuberance of CO2 and depletion of nitrogen [85].

The supply of energy from the transfer of photosynthetic electrons or extrinsic organic carbon

greatly affects the nitrite and nitrate reduction. The precise process by which NOx is ingested is

till now unknown. Rising the NO concentrations is not really well understood how will affect the

growth of microalgae. However, a study found that when having a high concentration of NOx

contained in the flue gas utilized for cultivation, a quicker rate of growth and a higher density of

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cell will be obtained. Other study showed that ranges between 17.65 and 88.25 mm have no

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impact on the photosynthetic efficiency and the growth of Chlorella sp. It was also observed that

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the growth rate decreased with higher concentrations of nitrite. One of the observations indicated
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that the growth of N.salina or P.trocornutum did not get affected by the NO [84].
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SOx

Combustion of compounds of organosulfur, sulfur, and hydrogen sulfide form the sulfur oxide.
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The existence of catalytic ash content, the sulfur chemistry in the fuel and the temperature of the
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combustion are considered as the factors used in dividing sulfur into SO2 and SO3. Sulfur in the

flue gas consists of around 2 vol percent of SO3, which reacts fast with water to form H2SO4

[84]. Highly dissolved concentrations of SO2 and CO2 can impact the PH, thereby it must be

controlled or buffered [87]. Certain species can withstand the high levels of NOx and SOx such

as Scenedesmos sp. and Chlorella sp. [85]. However, Chlorella sp.KR-1 demonstrated an

elevated tolerance levels of CO2 around 100ppm and around 15 vol percent of CO2. And in spite

of the verity that Chlorella sp.KR-1 showed a high tolerance to SOx compared with other
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species. Its growth was totally inhibited at concentration above 150 ppm of SO2. However, a

study investigated that when the SO2 concentration in the medium was 50 ppm, the rate of

growing of the microalgae was not impacted. On the other hand, increasing the concentration up

to 400 ppm (PH<4) lead to inhibit the growth within 20 h [84]. It was tested that the flue gases

from coal fired power plants work on enhancing the growth of the microalgae [85]. A study had

examined the impact of aerating a cultivation system consisting of Chlorella sp. By using flue

gas flow coming from a power plant, coke oven and hot stoke in a steel plant. It showed that the

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flue gas contains around 15-80 ppm of SO2, about 8 to 80 ppm of NOx and from 24 to 25 vol%

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of CO2 [88]. Some research mentioned that the existence of SOx and NOx in flue gases may

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lead to inhibit the growth [80]. Many metals are essential for the growth of the biomass as Zn,
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Fe, Mg, Co, Ni and Cu [87]. The microalgae need low concentration of specific heavy metals for

the synthesis of enzymes as some studies had mentioned. However, high toxicity may be
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produced from the high concentration of some heavy metals as Hg, Cr and Pb. A study had
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studied the effect of 10 popular flue gas heavy metals which are (cd, Hg, Zn, As, Se, Cr, Ni, Cu,

Pb, and Co) on the productivity of the microalgae. The results showed an improving in the
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growth of biomass about 12 percent and rising the lipids rate by 61 percent [84]. However,
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having more concentrations of flue gas metals will adversely affect the lipid production and

growth [83]. A study compared the growth of algae culture under flue gas and under control

culture that used a mixture of air (11% (v/v) CO2) and CO2 for supplying the culture. However, it

has been noticed that the growth rate was higher under the flue gas [84]. It’s obvious that the

gasses are benefit to the environment footprint [83].

4. Biomass Measurements and Growth Rates

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Biomass concentration determines the carbon dioxide fixation, since an increase in the

concentration does not only provide higher biomass quantity per volume of culture, but also

increases the amount of carbon fixed. Algae grow at different growth rates, the growth kinetics

takes into account the different culture parameters [63],[58]. For microalgae, the growth occurs

through 5 phases:

1. Induction Phase: During this phase, the increase in cell density is slow when transferred

into liquid culture. This is due to the cell metabolism adaption to growth, such as the

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increase in the amount of enzymes that are involved in carbon bio-fixation and cell

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division.

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2. Exponential Phase: At this phase, the cell concentration increases logarithmically as a
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function of time (t): Ct = Co (18), where Ct and Co are the concentrations at time t
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and 0, respectively, and m = specific growth rate, which depends mainly on the specie,

temperature, and light intensity.

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3. Declining Growth Rate: The cell divides slowly, due to physical or chemical factors such
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as: nutrients, pH, light intensity, or CO2 that starts to limit growth.

4. Stationary Phase: The cell density remains constant during this phase, as a result of the
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balance between the limiting factor and growth rate.

5. Death or (Crash) Phase: The algal cell density decreases, and factors such as: lack of

nutrients, overheating, and unbalanced pH cause algae crushes. The exponential phase or

(Growth Phase) determines the success of biomass production [71].

The biomass productivity is the increase in weight per unit time [58]. The maximum biomass

productivity (Pmax, g ) can be calculated from:

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Pmax = (15)

where is the biomass concentration (g at the end of cultivation time (day), and is

the initial concentration (g at (day) [89].

As the number of cells increases, the biomass production increases, which will lead to more lipid

yield [68].

The specific growth rate ( , ) is calculated from:

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(16)

where is the cell concentration at the beginning

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the exponential growth phase [89]. The cell growth can be indicated by measuring the
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dryweight of biomass, however, sometimes salt crystals add to the weight. Thus, determining the

chlorophyll content is considered a better option for growth measurement [63]. The growth of
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macroalgae begins with a rapid initial phase, then a decrease in growth occurs in later stages.

This is because the inner parts are often shaded from light subjection, thus reducing the
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photosynthesis rate. The growth rate of macroalgae is measured from the formula:
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Growth Rate (% [( ) ] (17)

Where is the initial weight, is the final weight, and t is the time (days) [90].

For microalgae, the growth can be detected by using several methods such as, cells counting in a

volume of culture, or using the absorbance or light scattering methods. However, for macroalgae,

optical methods cannot be applied since it requires a uniform suspension. The dry weight (DW)

measurement is often used for determining the growth rate, yet this method is achieved by oven
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drying, sun drying, or freeze drier, which includes sacrificing the biomass sample. Therefore, the

fresh weight (FW) measurement offers an alternative and accurate way that involves dewatering

the macroalgae using a variety of methods for biomass assessment [91].

5. Harvesting and Biodiesel Production

After the cultivation process, the biomass can be converted to biofuel, such as biodiesel or

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bioalcohol depending on the amount of accumulated lipids or sugars respectively. Biodiesel can

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be used to substitute mineral diesel. It is renewable, and less toxic compounds are released when

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burned. Some algal species are able to produce oil up to 60% of their weight to convert it to

biodiesel, thus, they are considered the most promising source for the production of biodiesel.
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The steps involved in producing biodiesel are: harvesting, dehydrating, oil extraction and
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conversion of oil into biodiesel [92].

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5.1 Harvesting Techniques

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Harvesting is known as extracting algae from its culture area. Dried process and other processes

for obtaining the needed products are obtained after harvesting the algae. Processing and
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harvesting are important in terms of mass production; however, these processes are expensive.

The method that will going to be used depends primarily on the final product and culture strains.

Some microalgae characteristic has impacts on the harvesting process as size and density. Size

and density of microalgae make them difficult in harvesting for biomass production, since

microalgae are very small and have low cell density ranging between 0.3-0.5 g/L, with few cases

reaching 5g, and its size is around 1 to 20 μm. The industrial scale is setting optimum
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requirements for cell sludge to contain at least 300-400g/L. That means suspended microalgae

should be concentrated at least 100 times, which is an energy-intensive operation [13].

Processing of algae is based on the concept of separating solid-liquid processes. One of the

critical stages of harvesting is by thickening algae suspension till a thick algal slurry or cake

forms. However, to allow for harvesting and processing, the water content should be reduced as

much as possible. Algae harvesting has two stages process (see Figure 11):

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 Bulk processing: removing algae from the suspension of bulk. Through this process, total

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solid matter can be around 2-7 percent, by the usage of flotation, gravity sedimentation,

or flocculation.


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Thickening: Using some techniques as ultrasonic aggregation, filtration, and
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centrifugation to concentrate the slurry. Therefore, this is usually a step more energy-
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intensive than bulk process.

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Figure 11: Drying and harvesting techniques [93]

Essentially, for viable production; the option of algae harvesting technology has to be energy-

efficient and relatively cheap. Extraction methods used is necessary for the final concentration of

slurry, which affects the energy input needed. Harvesting process includes thickening,

dewatering, and/or drying [7].

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Screening

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Screening is considered as the first process in most algae harvesting [7]. Screening is a type of

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operation used as the pre-processing of microalgae cultures [93]. The efficiency of it based on
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the distance between the openings screen on the panel and the cell size [7]. Vibrating screens and

microstrainers considered as the main instruments used for the screening [94]. Larger microalgae
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in this method are handled better with bigger openings, which conduct a lower operating costs
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and higher flow rates. Although the method's simplicity and low investment, it had been

demonstrated to have a poor efficiency in recovering microorganism sized of microalgae and

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further processing is needed.

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Thickening

Methods of thickening must be implemented to increase the concentration of the solid of biomass

suspension and to decrease the volume to be handled, because volume devaluation leads to

substantial savings in downstream procedure. Thickening processes generally compose of

flocculation/ coagulation (both biological and chemical), flotation, sedimentation by gravity, or

an electrical path [93].

Filtration
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Filtration uses suction-pumped membranes to isolate algae from water and suction pump is used

to force algae flow through the filter medium. The algae concentrated on the filter medium are

being harvested at the next step [71]. The benefit of this method is its ability for covering very

low density of microalgae cells [7]. Around 0.45 micron membrane filter is used for the algae

[71]. Regrettably, filtration can be clogged easily [95] and is comparatively having low

efficiency. In addition, although it’s simple, it’s too expensive [7].

Flocculation

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Flocculation allows algal cells to accumulate into larger clumps which are easier to filter and/or

settle faster, to allow their removal from downstream processes as seen in Figure 12 [94]. This
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technology is suitable for usage with a large range of microalgae which made it one of the most
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common microalgae harvesting process [96]. A study had determined that the energy
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requirement and cost for microalgae harvesting could be greatly decreased in case of flocculation

ability to pre concentrate the cell [97]. Flocculation alone is not that much efficient for
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harvesting in open pond systems of large scale. To enhance flocculation method, mixtures,
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concentrations, chemistry and size should optimize algae recovery depending on selection of the
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strain, algae flocculent interaction mechanisms, and in particular processes on empirical

determinations [7]. In flocculation method chemicals are used, to cause the algae to form lumps.

The downside of flocculation is that chemicals are utilized [97]. When algae are isolated,

chemicals are hard to extract from the algae. It is therefore not economically effective for

industrial use and for personal use and chemicals are costly for commercial use [71].
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Figure 12: Basic flocculation step [71]

Gravity sedimentation
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Due to the large volumes treated and low value of the biomass produced, gravity sedimentation
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is one of the most popular harvesting techniques for algae in wastewater treatment. It is used for

the separation of algae; since the purity of the overflow is primarily important and the suspension
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of algal feed is typically diluted [7]. The effectiveness of settling of solids by gravity depends on

the density and radius of the particles of the microalgae [94]. Particles of low density microalgae
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do not settle very well and cannot be effectively isolated by settling [7]. In this technique
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microalgae harvesting can be accomplished by means of lamella separators and sedimentation

tanks or ponds [94]. To boost the settling of algae, flat inclined plates are inserted into a settling

tank to facilitate interaction with solids and to settle them down the plates. The slope of the

plates let the settled algal particles to float down into the sump which they are drained from [7].

The solid content of algae concentrated into 1.6 percent [93]. This method is appropriate for

operational efficiency, but further thickening of the algae slurry is required [7]. Although the
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process's simplicity, sedimentation works for diverse microalgae strains and is highly energy

effective [93]. On the other hand, it’s considered as time consuming [98].

Stokes’ Law had described the sedimentation by making an assumption that velocity of

sedimentation is proportional to the square of Stokes’ cell radius and the density difference

between the microalgae cells and the medium as showing below:

(18)

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Where r is radius of the cell, η is dynamic viscosity of the fluid, is the solid density and is

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the liquid density [95].

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Floatation
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Flotation method is considered as an alternative to gravity sedimentation, which is specifically

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effective on suspensions of light algae. However, flotation is different from gravitational

sedimentation in a point; gravitational sedimentation works better with thick suspension of algae,
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while flotation can be used when suspended particles have such a low settling velocity that they
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cannot settle in sedimentation tanks [7]. Also it’s comparatively fast in compared with

sedimentation for a number of microalgae strains [95] and it’s more effective and beneficial [94].

Flotation is used for the algae harvesting in wastewater in conjunction with flocculation. The

algae floats on the medium's surface, and is eliminated as scum [71]. Flotation procedure is

categorized into various devices according to bubble size and production: electrolytic flotation,

dispersed air flotation and dissolved air flotation [95][94][96].

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• Dissolved Air Flotation — removes algae from medium by foam flotation and

flocculation. Using chromium, sodium, magnesium, titanium or potassium, an air

compressor with bubbles is provided and blended with air and algae.

• Froth Flotation — it works by separating algae from the culture by controlling pH level.

The air bubbles go through it to produce foam of accumulated algae above liquid level.

However, for commercial use it is too expensive [71].

The factors that may affect the efficiency of the flotation are ionic strength, the collector type

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(flocculant or surfactant), bubble formation type and level of PH in the medium [96]. One of the

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features of flotation is its ability in capturing particles with a diameter smaller than 500 [94].

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However, flotation has high operational cost and high energy usage, particularly if it needs small
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bubbles [95].
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Centrifugation
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Centrifugation is typically used in the separation of industrial suspensions, and many

investigations have been done on algal harvesting using this method. Centrifugation is a fast
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spinning container system that enforces centrifugal force to its contents, separating fluids or
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liquids from solid, as wastewater after treated and algae. The efficiency depends on the species

selected (as per size related). This technology is fast and energy intensive. Laboratory

centrifugation studies have shown that 80-90% of the microalgae can be recovered within 2-5

minutes. Even though, it is costly for individual use [7].

• The reverse flow vacuum system —can be used to prevent filter clogging. The pressure

comes from the top and the mechanism works softly to keep the cells from being packed.

It can be achieved in 3 hours using 20 liters to 300 ml.

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Harvesting methods Advantages Disadvantages

Chemical coagulation - Simple and fast method.
flocculation - No energy requirements.
Auto and bioflocculation - Inexpensive method.
- Allows culture medium
recycling.
- Non-toxic to microalgal biomass.
Gravity sedimentation - Simple and inexpensive method.
-Chemical flocculants may be expensive and
toxic to microalgal biomass.
-Recycling of culture medium is limited.
-Changes in cellular composition.
-Possibility of microbiological contamination.
-Time consuming

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• Direct vacuum —supplying a stir blade in the flask above the filter to keep the
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concentration phase from settling [71].

Table 2 summarizes the advantages and disadvantages of each of the harvesting methods

discussed.
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-Possibility of biomass deterioration.

Flotation -Feasible for large scale
applications
-Low cost method.
-Low space requirements.
-Short operation times.
Electrical based processes -Applicable to a wide variety of
microalgal species.
-Do not require the addition of
chemical flocculants.
Filtration -High recovery efficiencies.
-Allows the separation of hear
sensitive species.
-Low concentration of the algal cake.
-Generally, requires the use of chemical
flocculants.
-Unfeasible for marine microalgae harvesting.
-Poorly disseminated.
-High energetic and equipment costs.
-The possibility of fouling/clogging increases
operational costs.
-Membranes should be regularly cleaned.
-Membrane replacement and pumping represent
the major associated costs.

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Centrifugation -Fast method. -Expensive method.
-High recovery efficiencies. -High energy requirements.

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-Suitable for almost all microalgal -Suitable only for the recovery of high-value
species. products.
-Possibility of cell damage due to high shear

-p forces.

Table 2: Advantages and disadvantages of harvesting methods [93]

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5.2 Drying and dehydration

The two main challenges to implement an integrated algae system are by involving algae
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harvesting and a large-scale algae production in a way that permit biofuels and other bioproducts

to be produced for downstream procedure. Contamination utilized in some harvesting

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technologies would be difficult to remove, depending on the characteristics of the algae culture.
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The goal of increasing the productivity by controlling of the growth of the environment can

provide additional costs. Also enough land and water should be available [99].

Dehydrating:

After harvesting, dry process of algae slurry should exist for stabilization, extraction, end use and

other processes [7].

Drying
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Drying the flour algae requires a lot of efforts and creative methods [7]. Functional techniques of

drying algae would result in eliminating the algal degradation quality [100]. Drying methods are

mainly used for drying the slurry to a moisture content of around 12-15 percent [101]. There are

economic restrictions considered from the drying process on production of microalgae, It forms

around 70-75 percent of processing cost [102]. Different drying systems are distinct in terms of

energy demands and capital expenditure [7], especially in terms of energy consumption [100].

Moisture content of slurry microalgae should be reduced to at least 10 percent by using drying

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process and dehydration [13]. Final product and operation scale are the factors of selecting the

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proper method [101]. The objective is harvesting and drying significant amount of algae with

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reasonable cost [102]. It is important to mention that numerous of algal drying methods are
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advanced from conventional sewage sludge dewatering [100]. This paper will discuss some

drying methods used for drying algae.

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Drum drying
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Another name for drum drying is rotary drying, since it transfers materials being dried from one
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edge to another by sloped rotary cylinder using gravity. Some reports stated that using a thin
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layer drum dryer with a certain species give high dried algal product [7].

Solar heat drying:

This method is known as inexpensive and old drying form which can be easily achieved

by using solar radiation. Due to insufficient sunlight and unpredictable sunlight period, In

addition to the long time taking in drying and the large area required, solar heat is considered as

less effective method. Furthermore, unlike drum drying and oven, solar heat drying has no

sterilizing effects [13].

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Spray drying:

It comprises of atomizing the liquid, adding gas droplet with liquid droplet together. Droplet of

atomized water sprit down as well as hot gasses in a straight tower. Drying can be complemented

in a short time. Product after being dried is removed from downward and a cyclonic powder

separator drains the gas stream [7]. Spray drying is used for products with high value, however

the drawbacks of this method is the high costs and likely degrading the algae dramatically [13].

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Flash drying

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One of the popular methods in drying of wastewater sludge is flash drying, which has been

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developed in the US in the 30s century. This method was recommended for drying the algae,

since it provides quick moisture removal by using the spray way or by injecting a mixture of wet
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and dry algae through a hot gas stream [100]. The turbulent hot gases act as a transformer for
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moisture mass transfer from the algae slurry to the gases. Mainly, cost of the quality of the final
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algal product and the drying are significantly affected by the source of the hot gas. Unspotted

waste steam would help in reducing the working costs and guarantee having the final product in
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a good quality [101].

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Vacuum-Shelf drying

Depending on a study performed on Spirulina algae, it showed that drying of Spirulina slurry in a

vacuum shelf dryer with 50-60 °C and 0.06 atm [102], the slurry was dried to about 4 percent

moisture content [100]. The algae which have been dried set a porous structure of the biomass

and a hygroscopic characteristic [101].

Cross flow air drying

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A study showed wet Spirulina algae slurry contains about 55 to 66 percent moisture dried at 62

°C for 14 hours, using a cross flow air inside a compartment dryer which leads to produce high

quality dried algae product with thickness of 2 to 3mm and around 4 to 8 percent moisture

content [100]. The cell wall of Chlorella and Scenedesmus has been found to remain intact after

drying [101]. Additional evaluation showed that this method was quicker than solar heat drying

and less costly than drum drying [102].

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Toroidal dryer

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For drying the algae slurry, toroidal dryer was suggested. It works on the theory of jet milling

and does not involve moving of the parts. Transporting for drying region of solid algal biomass

is done by flow of high velocity air [101].

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Freeze drying
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It is well used in moderate operation conditions. One of its advantages is the ability of breaking
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and converting species cells into smooth powders, with no needing for homogeneity. On the

other hand, freeze drying need large investment and is very slow [13].
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Incinerator drying

For drying and burning wastewater sludge, multiple hearths incinerator manufactured with a

circular steel cylinder that composes of sundry hearths ranked in a vertical stack. The incinerator

can act as an individual dryer in case the input heats were decreased. Thus, the incinerator could

be utilized as a dryer for the algae, with the providence of hot gases. Both of flow of the hot

gases and the wet slurry flow down inside the furnace in parallel. In drying process, the

mechanism of the parallel flow of hot gases and the product is usually used to avoid burning or

scorching a sensitive heat substance as algae [101]. One more type of incinerator is called
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fluidized bed incinerator, it was initially utilized for sludge incineration. This device works on

using a fluidized sand bed as a heat source to foster a uniform algal solid combustion. The time

before the algal slurry is established, the fluidized bed used to preheat by using gas or fuel oil

[100].

Table 3 summarizes the advantages and disadvantages of drying methods. The main challenges

are generally linked to capital cost and high energy, with considering a good quality of products

[102]. Methods used to dry algae are still generally on small scale, primitive and traditional.

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Moreover, the algae industry is classified as an old industry [100].

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Drying methods Advantages -p Limitations
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Rotary drying -Dual advantage of sterilizing and -High energy cost to run dryer
disruption
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Spraying drying -Rapid and efficient drying -Degradation in product quality

method; appropriate for production due to high-pressure
of algae for human consumption atomization; high operating cost
and low digestibility of dried
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algae
Solar drying -The most feasible drying means in -Highly weather dependent with
remote vicinities lacking of energy problem of overheating and
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supply unreliable; not recommended for

algal products intended for
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Cross-flow air drying -Relatively cheap and fast drying
Vacuum-shelf drying -Efficient and fast drying
Flash drying -Rapid removal of moisture
Incinerator drying -Burning or scorching heat
sensitive algal biomass can be
prevented
Table 3: Advantages and limitations of drying methods [100]
human consumption
-High energy cost
-High capital and running costs
-Cost of drying and the final
product quality greatly
influenced by hot gas source
-Complicated and high capital
cost

5.3 Oil Extraction

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Many algae species accumulate neutral lipids in the form of triglycerides that have to be

extracted from the biomass for biodiesel production. Extraction algal oil has been done using

chemical and mechanical methods [33]. The oil yield efficiency is calculated from:

Extracted oil efficiency (wt. %) = (19)

1. Mechanical methods: These methods are categorized into mechanical expeller press and

ultrasonic extraction [33].

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 Expeller press: Pressing the oil is the simplest method used to extract the oil from

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the dry biomass. However, this method is considered less cost-effective, since it

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requires drying the algal biomass before pressing it out with an oil press [33], [59].
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 Ultrasonic extraction: This method is applied by creating bubbles in a solvent using
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ultrasonic waves. The cell walls will then break due to the shock waves caused by

the collapse of bubbles, releasing their contents into the solvent. This method can
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either be done using dry or wet algae. However, when using wet biomass, part of
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the water should be extracted prior to oil extraction [33].

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2. Chemical methods: Chemicals can efficiently recover the oil inside the algae, leaving

only 0.5-0.7% of leftover in algal biomass. This can be done using the solvent extraction

method, which is better than the mechanical pressing method. For this technique, hexane

is mostly used since it’s less expensive and toxic than other solvents like benzene and di-

ethyl ether [103].

 Hexane solvent method: This process can be combined with mechanical expeller press

method by squeezing out the oil using an expeller, then the remaining algae is mixed with
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hexane solvent for extracting all the oil content (See Figure 13). Finally, oil and hexane

are separated by distillation. Other solvents like ethanol (96%) and hexane-ethanol (96%)

mixture can also be used [33]. With this method it’s possible to extract 95% of oil from

algae [71]. A study was done to determine the effect of different parameters on the oil

extraction efficiency from dried and powdered algae using n-Hexane as a solvent. As the

Hexane to algae ratio increased, the oil yield efficiency has also increased. The extracted

oil yield was 2.5 times more when using 1:3 algae to solvent ratio than when using the

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1:1 ratio, this is due to the excess amount of solvent used to extract the oil from the

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biomass. Also, by using different algae biomass size and keeping the mass of algae and

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volume of solvent constant, it was observed that the contact area between the biomass
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and solvent has increased when using smaller size, thus, the extraction efficiency has also

increased. Lastly, by varying the contact time between the solvent and biomass from 10
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to 25 hours, while keeping other parameters constant, the percent of oil yield has
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increased as the interaction time was maximum, which enhanced the mixing and

solubility of oil [103].

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 [71][70][69][68][67][66][66][65][64][63] Soxhlet extraction: This method requires a special

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glassware equipment, called Soxhlet extractor, shown in figure 1 and 2. The oil is

extracted through repeated washing of algae using solvents, such as hexane or petroleum

ether as seen in Figure 14 [33].

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 Figure 13: Tube and Cylinder Soxhlet Extractor[63]

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[33][32][31][30][29][28][28][27][26][25]
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Figure 14: Soxhlet Extractor [25]

Supercritical fluid extraction: The supercritical carbon dioxide (scCO2) extraction requires CO2
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to be liquefied under high pressure and heated to achieve a supercritical phase at which a
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homogeneous phase that has the properties of liquid and gas appears. CO2 will then act as a

solvent to extract the oil [13], [33]. This technology is efficient and can produce a higher lipid

yield than the organic solvent extraction. The lipids extracted using wet Chlorococum sp. was

found to be higher than when using dry biomass, at 60 C and 30 MPa, with a resident time of 80

min, therefore, this technique minimizes the energy consumed during the drying process in

opposition to organic solvent extraction[13]. However, more energy is needed to achieve high

pressures [71]. Furthermore, the strain type and cultivation conditions can affect in determining

the most effective oil extraction method [13].

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5.4 Recent Advances in Oil Extraction from Algae

Many researches are being tested in order to develop better extraction methods. The single-step

extraction process is being developed so it can do the harvesting and oil extraction from algae

with separating it from water and the biomass in only one step in a short period of time. It is

efficient, uses no chemicals or expensive large equipment, and no drying of algae is required.

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Another technology has been developed by Cavitation Technologies Inc. (CTI), is continuous

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extraction by a Nano reactor that creates bubbles in a solvent that will further collapse near the

cells, creating shock waves that will break the cell walls and release the oil content into the

solvent. -p
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Oil extraction using mesoporous nanoparticles has also been developed to separate the high
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value compounds in the lipid mixture and then, the T300 catalyst was used for conversion of free
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fatty acids and triglycerides into algal oil and then to biodiesel. This conversion process is

efficient and cost effective.

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Hydrothermal liquefaction system has also shown successful results. It is based on the pressure
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cooker mechanism, which uses high temperature and pressure to convert algae slurry to oil. The

wet algae is heated to 350 C (622 F ) at a pressure of 3,000 PSI for 30 minutes while moving it,

then the algae breaks down and reforms into oil. The crude oil produced can be used for different

fuel applications. This process saves energy and cost used in drying the algae [71].

Oil refining
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Oil refining is when the crude oil that has been extracted from crops undergoes various refining

steps to remove waxes, phosphorus, free fatty acids and other impurities. The crude oil has to be

RBD oil before undergoing transesterification and producing biodiesel, which means the oil must

be refined, bleached, and deodorized. The refining process include the following steps:

degumming, neutralization, and dewaxing. In addition to bleaching and deodorization [19].

Degumming

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Dry gums or lecithin are unwanted phosphorous-related compounds in crude oil. Dry gums are

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generally removed during transesterification process. Lecithin is normally soluble in crude oil

but becomes insoluble upon hydration or in the presence of water. The gums can be removed by
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adding phosphoric acid or citric acid to the water, which also enhances the rehydration rate.
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After mixing and with sufficient contact time to hydrate the gums, the gums are separated by
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centrifugation or by a simple filtering process [19].

The ideal degumming process is one that efficiently controls the amount of water and acid while
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maintaining a suitable temperature. Furthermore, the length of contact time is necessary to

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achieve the maximum extent of degumming while reducing losses and improving product quality
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[19].

Water degumming, acid degumming, and enzyme degumming are the three common degumming

process techniques. Water degumming is the simplest degumming process, where hot water

between 60-80 C is used with about 1-3% of water used by oil volume [19]. In this process,

phospholipids absorb water and lose their lipophilic quality, becoming oil insoluble and

accumulating into gums, which are precipitated and then separated by gravitation or

centrifugation. Water degumming of vegetable oil are normally conducted by the addition of

water 2-5% by weight of oil. However, the phospholipids in algal oil are higher and cannot be
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removed with water less than 40%. The large amounts of phospholipids can inhibit the catalyst in

the transesterification reaction, resulting in a lower conversion rate, therefore the phosphorus

content should be reduced. Acid degumming process uses acids such as phosphoric, citric acid,

malic acid or EDTA. It provides higher phospholipids removal from mixed algal oil than water

degumming. Acid degumming consumes large amounts of water [20]. Enzyme degumming

process treats crude oil with lipases to solubilize the triglycerides. This process generates less

waste but with additional cost [19]. However, a simple enzyme degumming process is not

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preferable for high phospholipids containing oils [21].

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Neutralization
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Crude oils contain free fatty acids (FFA) from the natural hydrolysis of triglycerides. The FFAs
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are usually removed by adding an aqueous sodium hydroxide solution or potassium hydroxide

solution in the degummed oil, which converts the fatty acids into water soluble salts (soaps). The
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soap is then removed by draining or centrifugation. After removing the solution, the neutralized
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oil is washed with water in order to ensure the removal of all traces of soap; then it is dried

[104].
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Acid value is used to determine how much sodium hydroxide or potassium hydroxide is required

to neutralize FFA, beyond that needed for the transesterification process. It is calculated by the

following equation, if KOH is used.

Acid value = (20)

Where 56.1 is the molar mass of titrant (KOH), V is the volume of KOH solution required for the

titration in mL, M is the molarity of KOH solutions used, and S is the mass of sample.
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The percent FFA is calculated by the following equation:

% FFA = (21)

Where MA is the molar mass of the chosen acid [104].

Dewaxing

Oil crops such as sunflower and corn oils contain large amounts of waxes that must be removed

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in order to prevent the oil from clouding when brought to lower temperatures. The oil is usually

cooled to 5 C, mixed with water that contains a surfactant like sodium lauryl sulfate, and

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allowed to stand for few hours. When neutralization is carried at this temperature, the soaps wet

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the wax crystals and bring them into suspension in the aqueous phase; then it is removed by
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centrifugation or filtration [104].
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Bleaching
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Colored substances and oxidation products are removed by bleaching. Materials such as natural

clays (bentonites) treated with acids are used for bleaching oils to help improve their adsorptivity
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and filterability. Proper bleaching with acid-activated earth is a critical step in oil processing. If
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done correctly, the oil coming out from the bleaching press is almost colorless with a peroxide

value of zero. The bleaching earth must be entirely removed since clay acts as a strong oxidation

agent that can foul the downstream processing equipment. Moreover, it must be performed by a

good filtration facility. The recovered bleached oil should be protected from thermal and

oxidative abuses, because the oil in this condition is in its least stable state. Inert gases such as

nitrogen are generally used to fill tanks’ headspace of bleached oils in order to exclude oxygen

from the system. The purpose of oil bleaching is not just to remove the coloring compounds
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(chlorophyll, carotene) by adsorption, but also to remove peroxides and secondary oxidation

products from the oil [104].

The following are the three types of bleaching processes:

 Batch atmospheric: A temperature of around 104 C is used, and the bleaching

earth is added from the top of the tank with the agitator running. Afterwards, the

temperature is increased to the suitable bleaching temperature. Finally, the oil is

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circulated through a filter press [104].

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 Batch vacuum: A temperature of around 104 C is typically used. Slurry is made

in a tank and transferred to the vacuum bleacher. After completing the specified
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time at the bleaching temperature under vacuum, the oil is cooled, the vacuum is
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broken, and the oil is filtered [104].
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 Continuous vacuum: Bleaching clay is continuously fed into an oil stream at 104

C and sprayed into a vacuum chamber to remove water and air from the clay and
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the oil. After the temperature is increased, the oil is sprayed into a different
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chamber for bleaching. Then, the oil is cooled and the vacuum is broken [104].
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Deodorization

The last step in the oil refining process is deodorization. It is a vacuum-steam distillation process

at high temperature in which FFAs and volatile components are removed to get a bland and

odorless oil. In order to remove excess FFAs, the deodorizing temperature is usually increased to

250 C; however, this results in a higher production of trans fatty acids (1-3%) as well as a

higher loss of tocopherols. Moreover, deodorization removes odor and flavor-causing

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compounds. These volatile compounds can be removed by bubbling live steam through the hot

oil (200-275 C) under high vacuum (3-10 mm Hg). Deodorization does not have any significant

effect upon the fatty acid composition of most oils and fats. So, this process is not a necessary

step in biodiesel production. Furthermore, deodorization removes FFAs but the live steam causes

hydrolysis. The optimal conditions are 0.02-0.04 FFAs and 0.3-0.5% monoglycerides. The oil at

the end of this step is referred to as refined, bleached, and deodorized (RBD) oil [104].

The following is a set of pretreatment criteria for oil refining recommended by Desmet Ballestra

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[104]:

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1. FFA less than 0.10% (1000 ppm).

2. H2O less than 0.05% (500 ppm). -p

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3. Unsaponifiables, includes cholesterols, polymers, sterols, etc., less than 1%.
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4. Soap less than 0.05% (500 ppm).

5. Phosphorus less than 20 ppm.

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6. Sulfur less than 15 ppm.

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7. Impurities (sediment) less than 0.10% (1000 ppm).

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8. Breakdown fatty acid-sterol-glucose molecules.

Transesterification

The transesterification process is illustrated by the following schematic of Figure 15.

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Figure 15: Schematic of oil transesterification process [104].

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The refined, bleached, and deodorized oil is used to start the transesterification process. Many

experts suggest pretreatment of oil before producing biodiesel [104] as seen in Figure 16.
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Figure 16: Mass balance for the transesterification process [104].

For every 100 kg of RBD oil used, around 10 kg of methanol is used with the catalyst, resulting
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in the production of 100 kg of biodiesel and 10 kg of glycerin [104]. Triglycerides react with

methanol or alcohol in the presence of a catalyst such as sodium hydroxide, producing a mixture

of methyl esters and glycerin [105]. A homogeneous base catalyst such as potassium hydroxide

or sodium hydroxide is normally used to accelerate the reaction. However, base catalyst can react

with FFA in algae lipids to form soap, which results in a lower biodiesel yield as well as

increasing the difficulty of separating biodiesel from glycerol. Nonetheless, base-catalyzed

reactions are way faster than acid-catalyzed reactions that uses sulfuric acid as an alternative

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option. Base catalyst reactions are generally optimized at 60 C under atmospheric pressure for

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90 minutes. The operating cost is higher at higher temperatures and pressures. The biodiesel is

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separated from contaminants in a flask separator [13]. Furthermore, the crude methyl ester is
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washed to remove traces of methanol, glycerin, and catalyst; then it is dried to obtain biofuel.

The obtained biofuel is usually stored in tanks with nitrogen coverage in order to prevent
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oxidation [105].
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6. Conclusion
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As the demand for energy is always increasing, so is the use of fossil fuels, which results

in carbon emissions. This increases the need for a commercially viable carbon conversion

technology. The three major methods of CO2 capture (membranes, solvents, adsorbents) were

identified. The use of algae in CO2 capture and sequestration during the photosynthesis process

was explained. Algae cultivation system has provided a way to capture carbon and use it to

potentially produce biodiesel. Open systems are commonly used because they are simple and

cheaper, however, closed systems are better because they have a controllable environment, thus a

higher biomass productivity. Several parameters affect the CO2 fixation efficiency, including
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type of algae strain, light sources, nutrients, pH, temperature, and mass transfer. The impact of

flue gases composition, namely CO2, NOx, and SOx, on algal biomass was investigated.

Biomass measurements and growth rates are important in determining carbon dioxide fixation.

Harvesting and biodiesel conversion processes were identified and reviewed, which include

harvesting techniques, dehydrating, oil extraction, oil refining, and transesterification. The

presented technology and processes play an important role in the global effort towards a more

sustainable future.

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CRediT author statement

Abdul Hai Alami: Conceptualization, writing draft, Methodology, Data curation, preparation, writing-
reviewing and editing, supervision
Shamma Alasad: Writing draft, writing-reviewing and editing
Mennatalah Ali: Writing draft, writing-reviewing and editing
Maitha Alshamsi: Writing draft, writing-reviewing and editing

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Declaration of interests

☒ X The authors declare that they have no known competing financial interests or personal
relationships that could have appeared to influence the work reported in this paper.

☐The authors declare the following financial interests/personal relationships which may be considered
as potential competing interests:

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Graphical abstract

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Highlights
 Micro and macro algae are investigated for CO2 capture
 These algae are also considered as a biomass resource
 Various cultivation, CO2 sorption and harvesting techniques are discussed

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