Renewable and Sustainable Energy Reviews xxx (xxxx) xxx–xxx

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Renewable and Sustainable Energy Reviews

journal homepage: www.elsevier.com/locate/rser

Bioremediation of textile wastewater and successive biodiesel production

using microalgae
Tahir Fazalb, Azeem Mushtaqb, Fahad Rehmanb, Asad Ullah Khana,b, Naim Rashidb,
⁎ ⁎⁎
Wasif Farooqc, Muhammad Saif Ur Rehmana,b,e, , Jian Xud,e,
a
National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China
b
Department of Chemical Engineering, COMSATS Institute of Information Technology, Lahore, Pakistan
c
Department of Chemical Engineering, King Fahad University of Petroleum and Minerals, Dhahran 31261, Kingdom of Saudi Arabia
d
School of Chemistry and Chemical Engineering, Anhui University of Technology, Ma’anshan, China
e
Biochemical Engineering Research Center, Anhui University of Technology, Ma’anshan, China

A R T I C L E I N F O A B S T R A C T

Keywords: Microalgal biodiesel has emerged as an environment friendly alternative to the existing fossil fuels. The com-
Microalgae mercial production of this biodiesel is still challenging due to several technical and economic issues, which span
Textile wastewater from mass cultivation of microalgae to the biodiesel production. Mass cultivation is the most critical step in
Bioremediation terms of water and nutrient requirement. Industrial wastewater such as textile wastewater (TWW) is a cheap
Biodiesel
source for water, which additionally contains necessary nutrients (phosphate, nitrates, micronutrients etc.) and
Sustainability
organic dyes (potential carbon source) for algae cultivation. The application of microalgae for biodiesel pro-
duction employing single objective strategy is not sustainable. Microalgae can be effectively employed to
bioremediate TWW (dyes and nutrients removal) and to produce biodiesel from grown microalgae. This process
integration (bioremediation-biodiesel production) can potentially improve biodiesel production and wastewater
treatment. However, this process coupling needs to be thoroughly investigated to identify and optimize critical
process factors (algal species, cultivation and harvesting methods, bioremediation mechanism etc.). This study
has reviewed the status of TWW as potential source of water and nutrients, role of different algal species in the
bioremediation of TWW, different cultivation systems, harvesting and biodiesel production methods. This review
also suggests future research and development challenges for coupled textile wastewater treatment and mi-
croalgal biodiesel production.

1. Introduction Textile industry is one of the most important industrial sectors of

The growing human population has posed several challenges to the
global economy particularly in terms of environmental conservation and
energy security. Global economy is mainly relying on non-renewable and
finite fossil fuels [1–4]. This overuse of fossil fuels has resulted in two
highly correlated challenges of environmental pollution and energy in-
security. These fossil fuels have considerably contributed to greenhouse
gas (GHG) emissions where CO2 level has approached up to 400 ppm
[5–8]. Besides the use of fossil fuels in vehicular emissions, they are also
use in diesel generators to produce electricity for small and medium en-
terprises (SMEs) in developing countries like Pakistan. This excessive use
of fossil fuel (i.e. diesel) does not only cause financial liability to the local
industry. Thus, local industry needs to look for alternative strategies to
make their production cost competitive and environment friendly.
Pakistan that contributes a huge financial share. Recently, this in-
dustrial sector has confronted energy shortage that caused financial
loss. This industry switched their energy sources from national grid to
in-house diesel run generators to meet energy requirements. Although
this strategy has partly worked and provided continuous energy to the
industry but it also added financial liability. In order to save local
textile industry, there is an immediate need to search for sustainable
energy sources. Besides energy supply, textile industry is also facing the
challenge of wastewater management. Textile industry consumes sev-
eral hundred thousand gallons of water each day, and proportionally
produces huge volume of wastewater [9]. Textile wastewater contains
variety of dyes and auxiliary chemicals which may pose serious risks to
the environmental recipients [10–14]. The global concerns of energy
demand and non-tariff bindings regarding environmental conservation

⁎
Corresponding author at: Department of Chemical Engineering, COMSATS Institute of Information Technology, Lahore, Pakistan.
⁎⁎
Corresponding author.
E-mail addresses: drmsrehman@ciitlahore.edu.pk (M.S.U. Rehman), jianxu@ahut.edu.cn (J. Xu).

http://dx.doi.org/10.1016/j.rser.2017.10.029
Received 17 February 2017; Received in revised form 22 August 2017; Accepted 26 October 2017
1364-0321/ © 2017 Elsevier Ltd. All rights reserved.

Please cite this article as: Fazal, T., Renewable and Sustainable Energy Reviews (2017), https://doi.org/10.1016/j.rser.2017.10.029
T. Fazal et al.
may risk the fate of textile exports of Pakistan. Local textile industry
contributes 9% to the global textile needs [15] and more than 60% to
the total exports of the country [16]. Textile sector earned $13.7 billion
in 2013–2014 which dropped to $12.5 billion during 2015–2016 [17]
mainly due to energy liabilities.
Textile wastewater treatment is a big challenge because it contains
organic dyes, phosphates, nitrates etc. which cause multiple and multi-
scale damage to the receiving environment [18,19]. Pakistan require
textile wastewater treatment and reclamation to meet its domestic and
industrial water requirements [20]. Thus, sustainable energy supply
and textile wastewater treatment appear the biggest challenges to Pa-
kistan textile sectors [21,22]. In order to meet energy supply, textile
industry has already installed diesel run electricity generators. These
generators require financial liabilities in terms of both diesel con-
sumption and CO2 emissions. However, textile wastewater treatment
still remains unaddressed challenge. CO2 emissions can be manage
employing direct carbon capture, indirect carbon sequestration and
biological carbon mitigation techniques. Textile wastewater effluents
can be treated using several physico-chemical methods [23–29]. These
physico-chemical processes inherit several drawbacks such as ex-
pensive, less efficient, limited application and sludge handling
[8,30,31]. On the other hand, biological methods are cost effective and
environmentally friendly [14]. Though biological processes are attrac-
tive, but microorganisms (bacteria and fungi) require additional carbon
source for their growth to treat colored wastewater [32,33].
Recently, microalgae have received great attention due to their
potential to fix CO2 and bioremediation of textile wastewater [4]. Mi-
croalgae can simultaneously be used first for the remediation of textile
wastewater and later can be used as a feedstock for the biodiesel pro-
duction [34]. Thus, biodiesel can be used as an eco-friendly fuel in the
generation of electricity and can fulfill the energy requirements of
textile industry. Microalgae can use atmospheric CO2 (from diesel
generator) as carbon source along with organic dyes, and convert them
into carbohydrates through photosynthesis [35]. These carbohydrates
are converted chemically and biochemically into lipids which can be
used to produce biodiesel [36]. Microalgae biomass can contain up to
85% of total lipid content in dry biomass [37]. Therefore, microalgae
biofuels have potential to replace fossils fuels [4,38]. Microalgae are a
potential feedstock for biofuel production due to the requirements of
less amount of water for the growth as compared to terrestrial crops
[39]. Microalgae have also been reported to remove CO2, nitrogen,
phosphorus, and toxic metals from a different type of wastewaters
[40,41]. There are extensive studies available about the microalgae
cultivation in industrial wastewaters [42,43]. These organisms are
photosynthetic and categorized under third generation biofuels. Dif-
ferent species of microalgae: Chlorella vulgaris, Chlorella pyrenoidosa,
Spirogyra sp., Oscillatoria tenuisin and Scenedesmus sp. have shown their
capability to remove reactive dyes from textile wastewater [44,45].
Microalgae can be used to produce biodiesel [46–48], residual micro-
algae as fertilizer [49], and as cultivated biosorbent to treat wastewater
[50,51]. Microalgae may simultaneously solve all of the problems of
local textile industry as proposed in Fig. 1.
This review targets to overview simultaneous bioremediation of
TWW and the production of algal biomass, which can be useful in bio-
fuels production. This review elaborates the role of microalgae in the
bioremediation of TWW via biological and adsorption pathways, classi-
fication and short description of the cultivation methods, critical eva-
luation of microalgae cultivation, harvesting and oil extraction processes.
It further discusses about future R&D perspectives about bioremediation-
biodiesel process integration using microalgae and TWW.
2. Literature review
2.1. Textile wastewater characterization
Textile industries consume huge volume of water for their
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Renewable and Sustainable Energy Reviews xxx (xxxx) xxx–xxx
operations and thus, proportionally generate wastewater. Textile was-
tewater accounts for 30% (288,326 million gallons) of total wastewater
(962,335 million gallons) in the Pakistan [20] according to literature,
different sectors produce wastewater annually [52]. Unfortunately,
only < 8% of this wastewater gets treated prior to its disposal [20,53].
Textile wastewater (TWW) mainly contains high concentration of dif-
ferent dyes [45]. Globally, more than 100,000 different dyes are used in
different industrial sectors [54–57]. These dyes get discharged into
wastewater to a variable extent (10–60%) [11,57–61] and cause wa-
stage of 280,000 t dyes per annum [62–64]. The release of dye waste-
water may pose several risks to the receiving ecosystem [18,65,66].
Textile wastewater is characterized in terms of strong color, salinity,
temperature, pH, biological oxygen demand (BOD), chemical oxygen
demand (COD), total dissolved solids (TDS), total nitrogen (TN), total
phosphorous (TP) and non-biodegradable organic compounds
[18,19,60]. TWW also contains trace heavy metals like Chromium (Cr),
Arsenic (Ar), Copper (Cu), and Zinc (Zn) as presented in Table 1 [67].
The concentrations of nutrients vary in textile wastewaters is source
dependent [68]. For instance, TN ranges from 21 to 57 mg L−1 and TP
varies from 1.0 to 9.7 mg L−1 in textile wastewater [41]. In addition,
COD and BOD also vary due to the dyes used and their metabolites
produced in wastewater because different dyes inherit different struc-
ture [69–73].
2.2. Bioremediation of textile wastewater
TWW has complex composition containing dyes, salts, heavy metals,
reagents etc. [78–81]. Several processes have been investigated such as
adsorption with activated carbon, flocculation, ion exchange, mem-
brane filtration, electrochemical-destruction, ozonation and irradiation
[11,25,36,82–87], however, there is no standalone process, which can
treat TWW completely. Most of these processes are costly, energy in-
tensive, low efficient and produce sludge etc. [31,45,88].
Bioremediation has emerged as a potential technology to treat
textile effluents [33]. Different microorganisms (bacteria, fungi, yeast
and algae) have been explored for the bioremediation of textile dyes
[79,87,89]. However, microalgae is promising than other microbes
because it cannot only treat textile wastewater by uptaking nutrients
and dyes, but it can also accumulate lipids which can be trans-esterified
into biodiesel [90–92]. Biodiesel is an environment friendly sustainable
output of micrloalgal bioremediation process [93–96].
2.2.1. Micrloalgal bioremediation of TWW
Microalgae can be cultivated in textile wastewater, which use dyes
and nutrients for its growth. Microalgal bioremediation of TWW may
occur in two ways i.e. bioconversion or bioaccumulation process and
biosorption process. During the bioconversion process, microalgae
consume dyes as carbon source and convert them into metabolites.
However, microalgae also work as biosorbent where dyes can adsorb to
its surface. Both these phenomena can occur simultaneously for TWW
bioremediation [59]. The mechanism of microalgae accumulation can
involve enzyme degradation, adsorption or both [63] Additionally,
dead and living microalgae also participate in these phenomena [91].
However, the dead microalgae can only participate in the adsorptive
removal of dyes [77,97]. Microalgae result in high sorption capacity
due to their high surface area and strong binding affinity towards azo
dyes [98]. Microalgae species such as Chlorella vulagris, Chlorella pyr-
enoidosa and Oscillatoria tenuisin degrade azo dyes into simple aromatic
amines and decolorize dye wastewater [44,99]. Cheriaa et al. [100]
isolated new Chlorella alga and cultivated it in different textile dyes.
This alga decolorized different dyes variably such as indigo (89.3%),
direct blue (DB= 79%), remazol brilliant orange (RBO = 75.3%) and
crystal violet (72.5%) [100]. El-Kassas and Mohammad [101] culti-
vated Chlorella vulgaris in textile wastewater, and observed that it could
reduce COD up to 70% [101]. Another study found that Chlorella vul-
garis could degrade 63–69% mono-azo dyes into simple aromatics
T. Fazal et al. Renewable and Sustainable Energy Reviews xxx (xxxx) xxx–xxx

Fig. 1. The conceptual layout of microalgal-advanced oxidation process (MAP) for simultaneous textile wastewater treatment and biodiesel production.

Table 1 use light energy and carbon dioxide to produce biomass [118]. Thus,
The characteristics of textile wastewater [18,29,60,74–77]. microalgal cultivation becomes CO2 sequestration process as given in
Eq. (1) [119]. However, high CO2 concentration above certain limit
General characteristics Heavy metals
affects the biomass production and lipids accumulation [120,121].
Characteristics Units Value Characteristics Units Value
2H2O + CO2 + solar light→Cn(H2O)nNP + O2 + H2O [122] (1)
pH 5.6–9.0 Pb mg/L 0.5
Temperature °C 35–45 Cd mg/L 0.3
Besides CO2 sequestration, microalgal cultivation has already shown
Conductance mS 6.89 Zn mg/L 1.0 promising results for nutrient removal (91%-N, 93.5%-P and 45%-N
COD mg O2/L 250–8000 Mn mg/L 2.0 and 32%-P) from wastewater [123]. Another study revealed that high
BOD mg O2/L 50–550 Cr mg/L 1.5 concentration of nitrogen and phosphorus produced high biomass yield
TSS mg/L 100–700 Ni mg/L 1.0
and the higher lipid productivity was obtained in the starvation phase
TDS mg/L 5000–10000 Cu mg/L 0.2
NH4+1 mg/L 50 Fe mg/L 0.2 [124].
NO2−1 mg/L 350 Ar mg/L 0.01 Microalgae species are generally sensitive to different types of
Total PO4−1 mg/L 4.0–12 Total CN mg/L 1.0 wastewaters with imbalanced nutrient profiles such as the presence of
SO4−2 mg/L 50–900 Free residual Cl mg/L 100–500 inhibiting pollutants in wastewater and deficiencies in certain essential
trace elements. Typically, Chlorella sp. and Scenedesmus sp. are more
robust against different types of wastewaters [125–127]. Therefore
substances [102]. Microalgae G23 was successfully cultivated in textile
microalgae cultivation is an existing technology for algal biomass from
wastewater and it obtained 0.5 g/L biomass yield [103]. The bior-
textile wastewater, as well as their pollutant removal, since two decades
emediation of TWW and dyes employing different microalgal species is
For instance, Chlorella vulgaris could have removed 63–69% dyes from
presented in Table 2.
textile effluents [59].
Similar to bioconversion, microalgae can also carry out biosorption
Microalgae cell walls consist of proteins and carbohydrates, which
of TWW. For instance, Spirulina platensis was used as biosorbent to re-
provide functional groups to bond with cations such as metals and basic
move reactive red 120 (RR-120) from its aqueous solution. It achieved
dyes [58,128]. Thus, it is necessary to know the composition of mi-
the maximum biosorption capacity of 482.2 mg g−1 removing 97% RR-
croalgae for its bioremediation potential. The composition of different
120 from the solution [61]. Scenedesmus quadricauda has been suc-
microalgal species is presented in Table 4. Moreover, the growth of
cessfully employed as a biosorbent to remove remazol brilliant blue R
microalgae biomass strongly depends on its type, nutritional modes,
(RBBR) [89]. The removal of malachite green (MG) had been reported
and the cultivation conditions
using the biomass of Cosmarium sp. [73]. In order to use microalgae for
the bioremediation of textile wastewater and to use its lipids for bio-
diesel production, there is a need to understand the complete process 2.3.1. Impact of types of algae on bioremediation
chain from microalgal cultivation to biodiesel production. There are approximately 50,000 species of microalgae present on
earth and have a variety of stains adaptable to various environmental
conditions [129]. Microalgae are eukaryotic or prokaryotic photo-
2.3. Microalgae cultivation in wastewater synthetic microorganisms having a unicellular or simple multicellular
structure [130]. Eukaryotic unicellular microorganisms are microalgae
Microalgae cultivation depends on several factors, which include include Chlorella, Dunaliella salina, Chlamydomonas and diatoms (Ba-
microalgae species, growth medium, type of reactor used/cultivation cillariophyceae), whereas prokaryotic unicellular microorganisms are
methods, light medium etc. Textile wastewater contains dyes (C cyanobacteria include (Cyanophyceae, Spirulina, Anabaena and Sy-
source), nitrates (N source), phosphates (P source) and metals (micro nechocystis sp.) [131]. Macroalgae or seaweeds are multicellular eu-
nutrients) which are the main ingredients necessary for microalgae karyotic algae include brown algae (Phaeophyta), green algae (Chlor-
growth [41]. Thus, textile wastewater appears as promising and cheap ophyta) and red algae (Rhodophyta) having defined tissues with
growth medium for microalgal cultivation [59,114–117] in Pakistan. specialized cells [46]. Diatoms such as Haslea ostrearia (silicon-rich
Different TWW nutrients significantly affect the microalgae cultivation algae) and prokaryotic cyanobacteria offer various opportunities for
(Table 3). Microalgae are unicellular photosynthetic organisms that can metabolic engineering and biotechnology [132]. The single celled

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Table 2
Bioremediation of textile dye wastewater by using microalgae.

Microalgae species Target Biosorption capacity (mg/g) Decolorization (%) Biomass productivity (mg/L.day) Nutrients removal References

Spirulina platensis RR−120 482.2 99 – – [61]

Chlorella pyrenoidosa MB 21.3 > 90 – – [98]
Chlorella Sp. MB – 99.9 – – [95]
MO 70
Scenedesmus quadricauda RBBR Active 68 – – – [89]
Inactive 95.2
Spirogyra Synazol reactive – 85 – – [45]
Chlroella vulgaris Tectilon yellow 2 G – 63–69 – – [102]
Caulerpa scalpelliformis Sandocryl golden yellow 39 95 – – [97]
C−2G
Elkatothrix viridis Basic Fuschin – 93 – – [104]
Spirulina platensis RB19 251.61 – – COD 85.6% [86]
Spirogyra Sp. DB – 80 – – [105]
Spirogyra RY22 0.0004 92 – – [106]
Volvox aureus MR – 45 – – [104]
Spirogyra Sp. Cr(IV) 14.7 30 – – [107]
Chlorella pyrenoidosa DR31 30.53 96 – COD 82.7% [108]
PO41 19.9%
Desmodesmus sp. MB, MG 98 – – [109]
Oscillatoria sp. TWW 76 [110]
Chlorella vugaris RTWW – – 3.08 per cathode Zn 98% [81]
Cr 80%
Chlorococcum vitiosum TWW – 13 0.0335 div COD 13% [111]
Neochloris sp Industrial TWW – – 0.109 COD 34.5% [112]
Chlorella pyrenoidosa TWW 20.8 80 8.114 NO3−1 82% [98]
PO4−1 87%
Gloeocapsa pleurocapsoides FF sky Blue – 90 3.34 – [113]
Chroococcus minutus Amido Black 10B – 55 2.04 – [113]
Chlorella sp. G23 TWW – 75 0.00005 COD 75% [103]
NH4-N 72%
Chlorella vulgaris TWW 77 0.0019 COD 69.9% [101]

eukaryotic algae are size around 3–30 µm [133] and cyanobacteria size Table 4
ranges 0.2–3 µm [133]. Algal pathogens may compete for azo dyes, Chemical composition of several species of microalgae (% of dry mass) [49,129].
organic carbon and other nutrients resources [134]. The different
Microalgae species Protein Carbohydrate Lipid Pigments Carotenoids
genera of microalgae such as Chlorella vulgaris, Chlorella protothecoides
and Scenedesmus species, remove high dye concentration, ammonia, Chlorella vulgaris 51–58 12–17 14–22 0.5–1.0 0.1–0.2
nitrates, phosphates and heavy metals from textile wastewater [135]. Scenedesmus obliquus 50–56 10–17 12–14
Specifically, Chlorella vulgaris showed high removal rates for inorganic Dunaliella salina 57 32 6
Spirulina maxima 60–71 13–16 6–7
nitrogen (60–86%) and inorganic phosphorus (78–87%) from waste- Chlamydomonas 48 17 21
water [60,130]. Thus, the selection of suitable microalgal specie is an rheinhardii
important task to choose such a candidate that may potentially pretreat Synechococcus sp. 63 15 11
textile wastewater as well as accumulate maximum amount of lipids for Chllorella 52 10–11 15
protothecoides
biodiesel production. The potential microalgae, which can be used for
cultivation in textile wastewater, should possess the given traits: growth
under extreme conditions; rapid growth rate; large size cell or colonial 2.3.2. Role of cultivation mode in TWW process
morphology; robust to environmental conditions; tolerance of shear 2.3.2.1. Autotrophic. Microalgae can synthesize carbohydrates
force; high cell productivity; high lipid accumulation etc. Table 5 pre- employing photosynthesis process and using CO2 in the presence of
sents the comparison of different microalgal species with respect to light [116,136]. Different algal species (Chlorella vulgaris, Chlorella
their biomass productivity and lipid productivity.

Table 3
Effect of textile wastewater nutrients on microalgae cultivation [126].

Nutrients Effects on algae growth

Carbon, C Essential component for microalgae growth. Glucose in heterotrophic conditions promotes rates of growth and respiration. However, high C concentration
reduces growth rate and inhibit if it exceeds certain limit.
Nitrogen, N One of the major nutrients required for microalgae growth. It is usually present in the form of nitrogenous compounds like ammonia, nitrates and nitrites that
activate many enzymes to produce oxygen
Phosphorous, P It is a key factor in the energy metabolism of microalgae. It is used to support the formation of nucleic acid, lipids and adenosine-tri-phosphate (ATP).
Inorganic phosphates play an important role in microalgae cell growth and metabolism through phosphorylation reactions.
Magnesium, Mg It is used in enzymes activation. It is the central atom in chlorophyll. It is also used to synthesize genetic material such as DNA and RNA.
Potassium, K It assists in osmoregulation, proteins synthesis and ion exchange in cell growth membranes.
Zinc, Zn It activates carboxylases for fatty acids synthesis.
Iron, Fe It is a key factor in protein synthesis like ferrodoxin and cytochrome. High concentration reduces the cultivation of microalgae.
Molybdenum, Mb The enzymes like nitrogenase and nitrate reductase contain it. Its deficiency inhibits the nitrogen absorption.
Chromium, Cr It acts as a photosynthesis inhibitor i.e. high concentration can affect the cell machinery responsible for oxygen release.
Copper, Cu It improves the biomass yield and oil content

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Table 5 contents [142,143]. Photoautotrophic produce low biomass (117.6 ±

Comparison between different microalgae biomass and lipid productivity. 1.9 mg/L.day) compared to heterotrophic mode (178 ± 1.9 mg/L.day)
[127–129]. Although heterotrophic mode yield high algal density and
Species Biomass Lipids Productivity References
Productivity (g/ (g/L.Day) lipid content [144], but the cost of alternative organic C source makes
L.Day) this microalgae cultivation economically unfeasible. Thus,
heterotrophic cultivation mode is suitable for textile wastewater
Ankistrodesmus falcatus 0.34 0.056 [161]
where organic dyes can be uptaken as C source.
Botryococcus braunii 0.03 0.055 [186]
Chlorella vulgaris 0.73 0.204 [161]
autotrophic 2.3.2.3. Mixotrophic. The mixotrophic mode is the combination of both
Chlorella vulgaris 0.074 0.011 [187] autotrophic and heterotrophic conditions for microalgae growth [145].
autotrophic Organic (glucose, glycerol, acetate and other organic pollutants) and
Chlorella emersonii – 0.010–0.05 [159]
inorganic (CO2) C sources are simultaneously assimilated in the
Chlorella vulgaris 0.227 0.061 [188]
mixotrophic presence of light by the autotrophic and heterotrophic modes [146].
Dunaliella tertiolecta 0.10 0.698 [189] Microalgae are also reduces releasing CO2 during respiration of
Nannochloropsis sp 0.68 – [190] autotrophic algae cultivation. For instance, Chlorella vulgaris shows
Chlorella protothecoides – 1.214 [159]
low biomass productivity (0.35 ± 0.0001 g/L.day) in autotrophic mode
Scenedesmus sp 0.0714 0.0095 [187]
[128] which increases (6.0–6.15 g/L.day) if high organic C source is
added [147]. Furthermore, Chlorella Sp. showed higher lipid content
Table 6 (35 ± 4.2%) and higher lipid productivity (0.01 g/L/day) than other
Different species of microalgae accumulate oil contents. cultivation conditions [141]. The mixotrophic cultivation requires low
light intensity, so it can save light energy costs. Consequently,
Microalgae specie Nutrient mode Oil content (% dry cell Reference mixotrophic system can get benefits from both autotrophic and
weight)
heterotrophic modes of cultivation, and minimize their individual
Chlorella vulgaris Autotrophic 38 [150] limitations. However, this mode still requires nutrient
Chlorella emersonii Autotrophic 63 [151] supplementation. The use of textile wastewater can overcome this
Chlorella protothecoides Autotrophic 23 [139] shortcoming because it contains all of the necessary nutrients for
Chlorella zofingiensi Autotrophic 54.5 [152]
microalgal growth.
Chlorella pyrenoidosa Autotrophic 30.9 [153]
Scenedesmus sp Autotrophic 21.1 [154]
Chlorella protothecoides Heterotrophic 55 [129] 2.3.2.4. Photoheterotrophic. Photoheterotrophic nutrition can take
Chlorella pyrenoidosa Heterotrophic 37 [155] place both in the presence and absence of light. The light and organic
FACHB−9
carbons are employed as energy sources in this mode. The light as the
Nannochloropsis sp. Heterotrophic 20 [141]
Scenedesmus sp. Heterotrophic 52.6 [156]
energy source is required latter as compared to the mixotrophic
Scenedesmus obliquus Mixotrophic 43 [157] cultivation [3]. The light and carbon source need at same time in
Auxenochlorella Mixotrophic 20.82 [50] photoheterotrophic mode [148]. Photoheterotrophic nutritional mode
Chlorella vulgaris Mixotrophic 42 [158] avoids the limitations of light dependency which is the major
Nannochloropsis sp. Mixotrophic 27.5 [141]
obstruction for gaining high cell density in large scale photo-
Chlorella sp. Mixotrophic 15.6–25.4 [159]
bioreactors because they used organic carbon as a sole source of
energy in the absence of light [50]. Therefore, it is more favorable
emersonii, Chlorella zofinhiensis, Chlorella protothecoides, Chlorella for the wastewater treatment and lipid accumulation. Along with the
pyrenoidosa, and Scenedesmus sp.) are found to grow autotrophically, ability to grow heterotrophically in nutrient-rich and organic
and accumulate oil in algal biomass [38,125,137]. Table 6 presents the environment, algae reduces toxins and heavy metals from wastewater
capacity of oil accumulation of differential algal species. Light intensity levels of several contaminant heavy metals are significantly reduced
plays a critical role in the cultivation of photautotrophic microalgae [149].
[138]. Chlorella and Scenedesmus species may tolerate different level of As mentioned above, all modes inherit advantages and dis-
light intensity. They have been grown under light intensity of advantages for microalgae cultivation. Algal biomass productivity de-
200 mmol m2s−1 [6]. The light penetration in the cultivation system pends on cultivations conditions. Therefore, nutritional mode must be
becomes a problem with the growth in microalgae. The higher density appropriate to ensure optimal lipids productivity. An ideal system must
of algal cells does not allow light to diffuse properly in to the reactor, have low capital, operational and maintenance cost, high productivity,
and incident light may get attenuated. At high cell density, the cells controlled cultivation parameters (pH, temperature, mixing rate) and
suffer light limitation. Besides light attenuation due to algal cell reliability.
density, this factor becomes critical for colored textile wastewater.
This attenuation challenge requires a special reactor design to improve 2.3.3. Cultivation systems
light diffusion [139]. Furthermore, the cost of algal harvesting may Recently, there are three different systems, which are used for mi-
increases due to the low algal cell density [140] in case of croalgal cultivation. These systems include open pond, closed pond as
photoautotrophical cultivation. photobioreactors, and hybrid systems [164,165].

2.3.2.2. Heterotrophic. The heterotrophic mode requires organic carbon
as a source of energy. Heterotrophic algae converts sugar (glucose) into
lipids in the absence of light [50]. Heterotrophic microalgae cannot use
atmospheric CO2 and light therefore it uses organic carbon (glucose,
glycerol and acetate) as energy source for their metabolic function
[140,141]. Heterotrophic cultivation has several other advantages
including elimination of illumination cost, simple operation, higher
growth rates and higher lipid productivity. These microalgae yield high
algal cell density (up to 111.48 × 106 mL−1) [140] and high lipid
2.3.3.1. Open ponds. An open pond system has been used for the
microalgae cultivation in past few years using wastewater nutrients
and atmospheric CO2 in the presence of light [161]. Open ponds are
generally 0.3–0.6 m deep and 0.8–1 m wide [6]. There are different
types of open pond design are used, but the most commonly used
systems include inclined systems, circular ponds and raceway ponds as
shown in Fig. 2 [38,162]. Inclined systems are designed to induce flow
of the algal culture suspension from the top to the bottom of the sloping
surface. Gravity force is used as a turbulence to ensure proper mixing of
microalgae. The cell density also increases due to the better mixing

5
T. Fazal et al.
Circular pond
Closed Photo-Bioreactor
Fig. 2. Types of open pond systems [38] and tubular PBR system for algal cultivation [173].
conditions. This type of system suits to Chlorella, Phaeodactylum and
Scenedesmus species because it can survive under repetitive mixing
environment [38,163].
Circular ponds and raceway ponds occupy an area of about 1 ha,
whereas extensive ponds can extend up to 200 ha. The circular ponds
contain a central rotating agitator to mix the culture. However, rotating
agitator becomes ineffective in large pond size (≥1 ha) and require
extensive energy for mixing. Raceway ponds are also known as high
rate algal ponds (HRAP). It contains paddlewheel for close-loop re-
circulation pathway to ensure better mixing of culture. Front channel of
paddlewheel is used to introduce nutrients and microalgae culture.
Large size raceway ponds are low cost but less productive microalgal
cultivation system [49]. For instance, Chlorella and Spirulina sp. have
produced low biomass productivity (17 g m−2 day−1) in raceway pond
compared to closed systems [164]. Open pond systems require low
construction and operational cost. However, it demands for large land
area, which may increase capital cost. In order to fix land cost, water-
logged areas can be a suitable option to construct open ponds for cul-
tivation. There are several factors which affect microalgal cultivation in
open pond system such environmental factors, weather conditions,
contaminants/predators (bacteria and fungi) and unwanted species of
algae which may grow on the same resources.
2.3.3.2. Closed pond. Contrary to open pond system, photobioreactors
(PBRs) are used to cultivate microalgae under controlled environment.
Complex chemical reactions take place, which convert CO2, H2O and
nutrients to an oil-rich algae biomass in the presence of light. It
provides a controlled microalgal cultivation environment in terms of
pH, mixing, maximum sunlight intensity, culture density and
temperature [160,165,166]. Various PBR systems are used including
vertical tank, helical tube, air-lift, horizontal tube, tubular, flat-plate
and vertical column photobioreactors. Tubular PBR are made using
linear tube of glass or plastic, and can be arranged like helical tubes in
6
Renewable and Sustainable Energy Reviews xxx (xxxx) xxx–xxx
Raceway ponds
outdoor environment as shown in Fig. 2. Flat-plate PBRs are
constructed by using rectangular transparent material containers,
which possess high surface area to volume ratio. Another
configuration of PBR known as column PBR that gives good mixing
and manageable cultivation. It is easy to construct and allows maximum
exposure of light [165]. Airlift PBR offers high mass transfer rate,
regular light/dark cycles and algal cells face low shear stress. Gas
bubbling is used for the mixing of culture which is carried out by
moving the culture between riser and bottom section of bioreactor
[167]. These PBRs are appropriate for microalgal cultivation because
they allow low accumulation of dissolved oxygen and efficient
photosynthesis. However, these reactors lack in temperature control
and hydrodynamic load management.
2.3.3.3. Hybrid systems. The hybrid systems combine the properties of
open pond and closed bioreactor systems. During the first cultivation
stage, the desired specie is cultivated in an open pond. During the
second stage, the open pond culture with preferred strain is cultivated
in closed system [168]. This hybrid system can expose the algal culture
to nutrient stresses and may increases the biomass productivity and
lipid accumulation.
2.3.3.4. Comparison. Open pond systems inherit certain limitations for
the cultivation of algae such as non-uniform light diffusion, evaporation
losses, temperature variation, poor diffusion of atmospheric CO2, poor
mass transfer, weather factors, and contamination from other algal or
bacterial strains [169]. Open pond systems also have low biomass
productivity due to inefficient mixing. Therefore, the closed PBRs have
been preferred for the growth of culture [170]. PBRs ensure controlled
environment than open ponds. Algal growth rate is usually higher due
to better control in PBRs, such as carbon supply, water supply, efficient
exposure to light, pH, optimal temperature, better assimilation regime,
low risk of contaminants and better gas (CO2) supply rate [171]. The
T. Fazal et al.
Table 7
Advantages and limitations of microalgae systems [6,161,174].
Production system Advantages
Open/Raceway Easy to build, relative cheap, Easy maintenance, good for mass cultivation
pond
Tubular PBRs High biomass productivity, suitable for outdoor mass culture and less
contamination chances
Flat plate PBRs Provide high illumination surface area, good light path, relatively higher
biomass productivity, low contamination of unwanted algae, lower power
consumption and low accumulation of dissolved oxygen
Column PBRs Easy to sterilize, good mixing, low fouling, High mass transfer capacity and low
photo-oxidation
Hybrid Ponds Better control on culturing biomass cultivation and less power consumption,
contamination can be prevented to obtain high biomass productivity
(1.5–4.0 gL−1) in closed systems [94,170]. However, cultivation in
PBRs requires extensive energy due to CO2 pumping and nutrients
mixing. The hybrid system provides better control for microalgae
cultivation with low energy input. However, these hybrid systems
require high initial and maintenance cost due to the combination open
pond and closed system [172]. The comparison of these cultivation
systems is presented in Table 7.
2.3.4. Nutrients Stripping
Carbon, nitrogen and phosphorus are the three essential elements
required for biomass growth. In order to used wastewater as a growth
medium for microalgal growth, wastewater should possess a suitable
nutrients profile as shown in Table 1. The production of each ton of
algal biomass requires at least 1.83 t of CO2, which may be fixed from
the atmosphere (10–50 times than terrestrial plants) and external in-
dustrial sources as soluble carbonates salts [6,137,167]. Therefore,
microalgae cultivation can mitigate tons of CO2 [41] and can produce
about 280 t of biomass per hectare per year [175].
Microalgae can uptake dyes and nutrients present in textile waste-
water and atmospheric CO2 effectively and efficiently to accumulate
lipids [176,177]. Microalgal cultivation requires light energy, essential
nutrients (carbon, nitrogen, phosphate and silicon), macronutrients
(sulfur, calcium, potassium, magnesium, chlorides and sulfates) and
micronutrients (Mn, Co, Cu, Fe, Mb, Zn, Cr [171,178]. These nutrients
are present in textile wastewater at a variable concentration. Some
micronutrients are present up to a lethal dose in textile wastewater
[177]. These nutrients play a critical role in biomass enrichment by
facilitating cell growth, maintenance and synthesis of different kinds of
lipids, carbohydrates and oils [171].
Microalgae assimilate nutrients and heavy metal ions into the cells
for its growth. Microalgae mostly consume inorganic CO2 as a carbon
source and this CO2 may change the pH of water [179]. CO2 firstly gets
absorbed into water where it forms carbonic acid (H2CO3), which dis-
sociates into bicarbonate ions (HCO3−1) and hydrogen (H+1) ions.
Carbonic anhydrase enzyme facilitates the conversion of CO2 into
HCO3−1. Microalgae utilize these bicarbonates for their cell growth and
lipids production [92,135]. Furthermore, bicarbonates ions dissociate
into carbonate ions and hydrogen ions. During the cultivation of mi-
croalgae, the concentration of bicarbonate ions decrease, whereas car-
bonate ions increase which further combine with water to form hy-
droxide ions. These hydroxide ions consequently increase pH as
explained in the following Eqs. 2,3,4 [130].
CO2 + H2O ↔ H2CO3 ↔ HCO3−1 + H+
HCO3−1 ↔ CO3−2 +H +
−2 −1 −1
CO3 + H2O ↔ HCO3 + OH
The nitrification process converts ammonium ions into nitrate ions,
and produce protons. Microalgae consume released oxygen (Eqs. 5–6)
Renewable and Sustainable Energy Reviews xxx (xxxx) xxx–xxx
Limitations
Requires more area, high evaporation loss, high CO2 diffusion to the air,
Poor light utilization, low biomass productivity, contamination of
microbial and local algae
High initial installation cost and toxic accumulation of oxygen fouling
overheating, large foot prints. Walls growth which affect light diffusivity,
Required large area of land, many support materials needed for scaling
and difficult temperature control
Small illumination surface area, scale up, difficult to construct, high
energy consumption
High initial investment cost and high maintenance cost
for its metabolic activity, which reduces aeration cost of microalgal
cultivation. Nitrogen plays an important role in lipid accumulation in
microalgae. Nitrogen stress has been found the only feasible and eco-
nomical technique to enhance lipid productivity on the large scale
[128,180–183]. Nitrogen starvation activates specific metabolic pro-
cesses in microalgae to store excess carbon into high energy molecules
such as lipids [184]. Nitrogen deficiency in growth medium helps mi-
croalgae to accumulate more lipids. On contrary, the mass transfer rate
of nitrogen limits the growth of algal cells due to lesser nutrients in the
medium as shown in following Eqs. (5) and (6).
1·0 NO3+ 5·7 (CO2) + 5:4H2O (C5·7 H9·8 O2·3 N1·0) + 8:25 O2 + 1·0
(OH−1) (5)
1(NH4) + 7:6 (CO2) + 17·7H2O→(C7·6H8·1O2·5 N1·0) + 7:6 O2 + 15·2
(H2O) + 1·0 (H+) (6)
Meanwhile, phosphates macronutrients are used to support the
production of nucleic acids, lipids and proteins [185]. The phosphates
get integrated into energy input organic compounds via phosphoryla-
tion. NADP+ and adenosine diphosphate (ADP) that get incorporated
into NADPH and adenosine triphosphate (ATP) respectively [6,41].
regulate microalgal metabolic pathways. The assimilation of nitrogen
and phosphorus depends on N/P ratio of wastewater. Different studies
reported that mostly Chlorella vulgaris and Scenedesmus sp. require op-
timum 8:1 and 5:1 to 20:1 N/P ratio, which may subsequently produce
high biomass yield [128]. The removal efficiency of these nutrients
from different wastewaters is reported in Table 8.
2.4. Microalgae harvesting
Microalgal harvesting is a critical step in the biodiesel production.
Microalgae culture is very dilute, which makes its harvesting a diffi-
cult task [168,196]. Separation of algal biomass from the suspension,
and its thickening are the main steps of harvesting [68]. Microalgae
harvesting is an important task within the biodiesel process because it
requires high energy input to harvest small sized microalgae (gen-
erally 1–20 µm) from a diluted culture [197]. The harvesting cost can
reach up to 20–30% of the total cost for biodiesel production
[170,198,199]. There is not a single standalone harvesting technique
available, which is efficient and cost effective. An ideal harvesting
technique should feature energy and chemicals efficiency, species in-
dependence, maximum harvesting efficiency and minimum lipid loss.
There are several factors which affect the choice of an appropriate
harvesting technique such as microalgae species, cell density, cell size,
(2) target products, harvesting efficiency and economical factor [116].
Available harvesting techniques include mechanical, chemical, bio-
(3) logical, electrical methods or a combination of two or more of these
processes. Several harvesting techniques have been investigated
(4)
which include centrifugation, gravity sedimentation, filtration, dis-
solved air flotation, flocculation/coagulation, screening and electro-
phoresis [131,200–202]. These techniques have been applied on lab
7
T. Fazal et al. Renewable and Sustainable Energy Reviews xxx (xxxx) xxx–xxx

Table 8
Nutrients removal efficiency with microalgae from different industrial wastewater.

Wastewater type Species Initial value TN (mg/L) Removal efficiency (%) Initial value TP (mg/L) Removal efficiency (%) Reference

Textile wastewater C. vulgaris 21–57 – 1.0–9.7 [191]

TKN
Textile wastewater C. pyrenoidosa 267 62 56 87 [60]
Agroindustrial wastewater C. Vulgaris 3–36 30–95 112 20–55 [192]
Piggery wastewater C. zofingiensis 16 – 7.0 [123]
Sago starch wastewater Spirulina platensis 2.0–3.0 96–100 18–21 87–99 [194]
Textile wastewater C. Vulgaris 45 33 [18]
Textile wastewater Spirogyra sp. 5000 95 5000 90 [193]
Cladophora sp. 5000 93 5000 88
Synthetic wastewater N. oleoabundans 1.2 99 0.03 100 [195]

Microalgae Harvesting

Filtration or Gravity
Centrifugation Electrolysis Flotation
Screening Sedimentation

Membrane
Filtration
Flocculation
Electrolytic Dissolved Dispersed
Ozone Electrolytic
Coagulation Air Air
Flotation Flotation
Flotation Flotation

Electrolytic
Flocculation Auto Chemical Bio
Flocculation Flocculation Flocculation

Harvesting Parameters

Settling Cell Cell

Air Mixing pH Temperature Flocculants
Time Size walls

Dosing Type
Zeta Potential
Bubble
Growth
size
Phase
Surface Polysaccharides
Charge Culture
Dissolved
Growth
Air

Fig. 3. Microalgal harvesting (a) Different types of harvesting methods (b) parameters which affect harvesting process [208].

scale, thus, their scale up is technically and economically challenging 2.4.1. Sedimentation
task. The promising harvesting techniques are discussed hereafter Sedimentation is one of the simplest and inexpensive harvesting
(Fig. 3). techniques. The biomass cells are separated using gravitational force of
the cultivation medium. Sedimentation is the most common harvesting

8
T. Fazal et al.
technique, which is not energy extensive and it is used for the settle-
ment of microalgae cells in large volume of wastewater. This technique
is effective for the harvesting of large sized and heavy cells. However, it
does not suit to small cells and requires very long time for their set-
tlement. The settling rate depends on species, cell size, cell density and
nutrient concentration etc. Poor light availability and low nutrient
concentration in the medium result in low settling rates. Average set-
tling velocity for green microalgae and diatoms is about 0.1 m day−1
and 0.2 m day−1. Sedimentation is used as a first step when combined
with other methods to optimize overall energy consumption of har-
vesting process [116].
2.4.2. Centrifugation
Centrifugation is a reliable, fast and efficient harvesting method,
which employs centrifugal force technique for the separation of mi-
croalgal cells from the growth medium. It results in 80–90% harvesting
efficiency within 2–3 min of operation [3,203]. Intense centrifugal
forces can damage the cell structure during the centrifugation process
[133,174]. The algal residue of Chlorella sp. was harvested by the
centrifugation at 3000 rpm, and was washed three times to get dry
biomass [204]. Different types of centrifuge are used such as nozzle
type, solid-ejecting disc, solid-bowl-decanter and multi chamber cen-
trifuges [201,205]. The choice of centrifugation process depends on cell
size and harvesting efficiency [206]. This mechanical process is energy
intensive, time consuming and requires high operational cost for large
volumes [39,202].
2.4.3. Filtration
Filtration is dewatering technology used for microalgae harvesting.
Filtration greatly depends on cell size. Different type of filtration
techniques are used for harvesting including vacuum filtration, dead
end filtration, membrane filtration (macro filtration > 10 µm), micro-
filtration (0.1–10 µm), ultra-filtration (0.02–0.2 µm) and reverse os-
mosis (<0.001 µm) [116,207,208]. During filtration, microalgae
growth medium is passed through a membrane/screen with a particular
pore size/mesh number. These membranes retain microalgae cells re-
tain and filtrate gets through it. The pore size of the filter depends on
microalgal specie. For instance, Chlorella sp. and Chlorella vulgaris with
the size of 5–6 µm and 0.1 µm can be harvested using microfiltration
[207,209]. The filtration of Neochloris sp. has been investigated when
the microalgae grown in dyeing industrial effluents and produced
36.69% of total lipids [112]. Nannochloropsis salina algas broth was
filtered using microfiber disc filter. This harvesting method resulted in
95% efficiency [208]. The filtration flux determines the efficiency of
harvesting. High flux rate does not provide the high harvesting effi-
ciency because the extracellular polymer substance (EPS) presence re-
duces it in microalgal growth medium [210,211]. EPS causes the
fouling/clogging of filter/membrane depending on surface charge, cell
size, cultural age and membrane type. Biofouling, energy requirement
and high capital and operation costs challenge the feasibility of filtra-
tion process [205]. Membrane fouling can also resolved by using
counter-current or turbulent flow, NaClO washing and other suitable
flowing velocities [212]. Besides several challenges, membrane har-
vesting of microalgae may become promising technique in future due to
massive R&D going on this field.
2.4.4. Flotation
Floatation is another harvesting technique where algal cells float on
the surface of the liquid due to air/gas bubbles. There are different
types of floatation based on bubble size such as dissolved air flotation,
dispersed flotation, electro flotation and ozone flotation. Dissolved air
flotation can be used for small sized and lighter algal cells harvesting
[205]. Bubble size (10–100 µm) is desired in dissolved air flotation.
This floatation depends on gas pressure, hydraulic retention time and
floating rates of microalgae cells [213]. Dispersed air flotation is
slightly different due to air bubbles size (700–1500 µm), where air
9
Renewable and Sustainable Energy Reviews xxx (xxxx) xxx–xxx
bubbles attach to the negatively charged surfaces of algal cells and
bring them to the liquid surface [214]. Ozone flotation disrupts the
microalgae cell walls and release proteins thereof. These released pro-
teins act as bio-flocculants and facilitate algal harvesting. Recently,
electro floatation has been widely employing hydrogen gas bubbles
which form during water electrolysis [202]. Flotation share several
advantages than other harvesting technique because it is simple to
operate, and require less time. On the other hand, it is controlled by
bubble size and involves high power cost to generate small air bubbles.
Recently, microfluidic oscillation was used to produce micro-bubbles at
low cost. This technique may reach 50–90% harvesting efficiency
[174].
2.4.5. Flocculation
Flocculation is a simple and cheap technology harvesting micro-
algae-colloidal suspension from water treatment processes [215,216].
Microalgal cells are typically <15 µm in diameter and their density is
slightly greater than water. Thus, flocculants or coagulants are added in
the microalgal suspension where these flocculants attach to negatively
charged surfaces of microalgal cells. These macro flocs (flocculants-
algal cells) settle down under sedimentation mechanism [41]. Floccu-
lation may result in 80–90% harvesting efficiency. For instance, in-
organic (FeCl3 and H2O2) flocculants could harvest Chlroella sp. Up to
90% [95]. Dunaliella tertiolecta biomass has been recovered with the
efficiency of above 90% [201]. Flocculation requires less energy com-
pared with other algal harvesting techniques such as sedimentation,
centrifugation and filtration [154]. Microalgae cells can be separated
from cell-floc complex by introducing electrostatically opposite charged
ions. Flocculants are divided into three major groups such as inorganic,
organic and bio-flocculants. The inorganic flocculants mainly consists
of multivalent cations (Al+3 and Fe+3) of aluminum sulfate
(Al2(SO4)3), Ferric sulfate (Fe2(SO4)3) and ferric chloride (FeCl3)
[168,201]. The type and dose of inorganic flocculant depends on algal
species. The dose may range from 10 to 50 mg/L or more. These in-
organic flocculants neutralize or reduce the zeta potential of microalgae
cells, and then facilitate their self-aggregation [39,133]. These flocs can
be harvested using other process such as sedimentation, centrifugation
and filtration. For instance, oxidized dye wastewater has been used for
the growth Chlorella sp and Fenton-like FeCl3 coagulants were used for
harvesting [95]. These inorganic flocculants also exhibit some limita-
tions such as pH sensitivity, specie specificity, pre cell lysis and the
contamination of harvested biomass with aluminum and iron salts
[198,217].
Organic or polymer based flocculants can also be used such as ca-
tionic starch, grafted starch, chitosan etc. These flocculants do not
contaminate the biomass and may be potentially nontoxic. Natural
polymers such as chitosan can be a promising alternative to address
these challenges [198]. It is nontoxic, biodegradable, high molecular
weight and cation charged density. It is cheaper than other flocculants
and result in high flocculation efficiency [201] and requires low dose
for harvesting. In flocculation process, electrostatically positively
charged chitosan cells attach to negatively charged microalgal cells to
form flocs [218,219].
The bio-flocculants such as fungi, enzymes and oleaginous microbes
can be co-cultured with microalgae [149]. It is an effective approach for
effective algal harvesting. Fungi can degrade cellulose of microalgae
cell, and pelletize microalgae to accelerate harvesting. Oleaginous mi-
crobes can also carry algal harvesting. These microbes follow two main
steps in this mechanism i.e. bridging (networking of microalgal cells)
and patching (bonding cells by EPS) during this flocculation. This
flocculation may give 80–98% harvesting efficiency [216]. Chlorella
vulgaris and Scenedesmus obliquus was harvested using Moringa oleifera
seeds with efficiency of about 80% and 92% respectively [220]. How-
ever, EPS produce more ions and increase the zeta potential of algal
cells Flocculation can be followed by mechanical methods to separate
the flocs from wastewater. The wastewater medium can be recycled
T. Fazal et al. Renewable and Sustainable Energy Reviews xxx (xxxx) xxx–xxx

back to the cultivation unit [201]. However, bio-flocculation is affected

[133,168,209]

[202,208,214]

[68,201,222]
by environmental conditions which are the most relevant aspects to

[168,201]

[216,223]
improve.

[3,170]

[3,68]
Ref
2.4.6. Electrolytic harvesting

Species depended (diatoms), Lack

Dose depended, impact on health

This method generates ions in microalgal suspension, and then it

Efficiency decrease due to EPS

Used for species specific, high
uses these ions to separate algal cells. This method has two further types

microalgae specie dependent

due to inorganic flocculants

Expensive, cake formation
such as electro coagulation/flocculation and electrophoresis.

Electrodes replacement,
Electrolytic coagulation is based on electrolytic oxidation of coagulants.
The positively charged coagulants formed complex with hydroxide

infrastructure cost
High capital cost

of proper design
ions, and attach with the negatively charged microalgae [198]. In
electrophoresis, there is no need for coagulants and flocs formation. The

Limitations
separation of algae cells is carried out in an electric field on the basis of
zeta potential of microalgae cells towards anode for the neutralization
[214]. Chlorella vulgaris grown in textile wastewater could be harvested

High cost due to high energy consumption, scaling of electrodes and low
required high area to volume ratio, dependent on air flow rate, hydraulic
high operating cost for small size bubble, expensive for ozone floatation,
at bio-cathode in single chamber microbial fuel cell yielding 3.08 ×
10−3g microalgal weight per cathode area [81]. Low harvesting effi-

Very long time for small uniform suspended culture (0.1–2.6 cm h−1)

Complex formation between bacteria and algae or formation of EPS

Operating cost is high due to flocculant chemicals, high final water
ciency has been achieved for marine microalgae medium due to the

Time consuming, fouling of filters, high operational cost due to

high ionic strength of water. This method is highly efficient and con-
fiscates about 80–95% of algal cells [214]. These techniques operate at
natural pH with low coagulant dose and short process time [81,168].

backwashing, presence of EPS reduce the efficiency

However, it requires high energy for large scale application, needs ex-
pensive electrodes, results in low efficiency for marine species [221].

content in the biomass slurry, cell disruption

Energy incentive, scaling up, Cell disruption

when no additional flocculants are presents.

All of the discussed harvesting techniques offer their advantages and

retention time and surface characteristics

disadvantage. Table 9 presents a comprehensive comparison of these
techniques.

efficiency for marine microalgae.

2.5. Lipids extraction

Harvested biomass is used to extract the lipid for biodiesel pro-

duction. Harvested microalgae contains lipid, carbohydrates, proteins
Disadvantages

and other valuable products [154,224]. Triglycerides (TG) and free

fatty acids (FFA), glycolipids and phospholipids can be extracted from
C. vulgaris, Scenedesmus ecornis and Dunaliella [225,226]. Lipids can be
converted into biodiesel through transesterification [34]. The residual
microalgae after lipids extraction can further be used to produce me-
Established and simple method, easy to control, residual stream can be

Air bubbles effectively interact with zeta potential of algal cells, DAF

thane gas from anaerobic digestion. High efficiency, low sludge volume, requires less energy and lower
It is hugely dependent on particle size, High efficient for large size

No coagulants / flocculants are used, rapid harvesting, working at

Reliable, fast and efficient as obtaining 80% biomass within 2–3

Cell drying and cell lysis are carried out using dry route or wet route capital cost, oxidant improve harvesting, and polymers do not
before oil extraction [227]. These steps require 2.36 and 0.62 MJ/L
biodiesel, respectively [121]. Thus, algal oil extraction becomes costly
and challenging step. There are four types of cell lysis techniques used,
which include mechanical, chemical, biological and supercritical fluid
microalgae, high fluid velocity reducing fouling

methods [226]. Oil extraction method should possess certain traits like
contaminate biomass as chemical flocculants.

fast, lipid specific, less lipid loss and minimum denaturation of cellular
Advantages, disadvantages and limitations of microalgae harvesting techniques.

material. Biodiesel production is not yet considered an economical

process due to the drying requirements. Therefore, another extraction
method has been developed that requires cell disruption instead of
is cheap at large volume scale

natural pH, required less time

It is an inexpensive method,

recycle to growth chamber.

drying [228]. The cellular material can deteriorate the conversion of

lipids oil into biofuel as shown in Fig. 4.
Highly effective,

2.5.1. Physical methods

Physical methods are involved in oil extraction by drying and cell
Advantages

lysis (cell disruption) of microalgae cells. Microalgae biomass can be

mints.

sun dried in tropical countries due to intensive sunlight [197]. Sun

drying is a cheap but time-consuming approach. It requires high surface
area, which leads to loss of valuable products. So, the efficient drying
Efficiency

80–90%

10–50%

70–95%

80–90%

70–90%

80–95%

methods include drum drying, freeze drying, fluidized bed drying and
99%

spray drying [229]. Cell lysis is carried out after drying step. There are
several mechanical methods used for cell lysis such as high-pressure
Electrolytic techniques

homogenization, microwave, ultrasonication, bead beating, electro-

poration, expeller pressing, glass beads, screw press extruder and pul-
Coagulation

Bio-flocculation
Flocculation or

verization [116]. The efficiency of physical methods depends on cell

Centrifugation

Sedimentation
Techniques

size e.g. Chlorella sp. cell wall is too rigid to disrupt Spirulina specie.
Filtration

Flotation

Physical methods can increase lipids oil concentration upto75% from

Table 9

dried cell weight [38,221].

Microwave (400–800 kHz) can disrupt the weak bonding in the cell

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T. Fazal et al. Renewable and Sustainable Energy Reviews xxx (xxxx) xxx–xxx

Fig. 4. Comparison of dry route and wet route of

lipid extraction from microalgae [228].
Higher Extraction yield
High energy consumptions

Dry Route

Drying Extraction

Second
Cultivation and First Step Biodiesel
Step
Harvesting Conversion

Cell
Extraction
Disruption
Wet Route
Much better energy balance
Low extraction yield
Effective cell disruption
required

wall. MV irradiations directly penetrate into biomass and disrupt the
cell rapidly [208]. For example, MV improved the efficiency of Chlorella
vulgaris cell lysis by 2.6 fold within 6 min [228]. Nannochloropsis sp.
was operated in microwave irradiation to disrupt cell (97.73%) and
biodiesel yield (36.79%) in the presence of organic co-solvent [230].
Ultrasonication uses micro-bubbles to disrupt cell walls and extract
components. These micro-bubbles create localized high temperature
and pressure, and intense shockwaves produced cause cell lysis.
Ultrasonic extraction is efficient but depends on specie, micro-bubble
size, cell density, temperature and frequency [187]. Bead beating is a
simple cell disruption method where beads (beads made of quartz or
metal) break cells by shaking closed container filled with algal cells.
The bead beating efficiency depends on various factors such as con-
tainer shape, shaking rate, bead size and number of beads used. This
technique is simple and it causes rapid cell disruption. However, it is
limited due to scaling up, energy and requirement for cooling system to
avoid thermal degradation. High pressure homogenization (French
press) method is more efficient to cell lysis for large scale operations
[47]. The hydraulic shear force is utilized for cell disruption. The
homogenization requires low energy, and does not pose high risk of
denaturing of cells.
2.5.2. Chemical methods
Chemical methods use polar (methanol) and non-polar solvents
(ethanol, 2-propanol, diethyl ether, hexane/cyclohexane, chloroform).
There are different types of chemical methods exist such as Soxhlet
extraction, accelerated solvent extraction, and Bligh and dryer methods
[226,231,232]. The repeated washing of microalgae is carried out
under reflux using organic solvents (hexane, benzene, chloroform, cy-
clohexane and acetone) in Soxhlet extractor. This method is cost ef-
fective and requires small volume of solvents. Alternatively, polar and
non-polar solvents combination can be used for efficient oil extraction.
Non-polar solvents disrupt the cell walls due to hydrophobic interaction
between neutral lipids and solvents. Polar solvents disrupt hydrogen
bonds in polar lipids [39]. Bligh and dyer method is more efficient
because it extracts more lipids than Soxhlet extraction [186,233].
Chemical methods inherit several disadvantages such as time con-
suming and contamination of fatty acids. These disadvantages can be
balanced employing an integrated approach i.e. mechanical-solvent cell
lysis for large-scale oil extraction.
2.5.3. Biological method
Enzymatic cell lysis disrupts the sporopolennin layer of algal cells
instead of whole cell. This method is selective and employs mild con-
ditions. Enzymes cleave specific chemical linkage and do not con-
taminate extracted fatty acids [228]. Pectinase and neutrase enzymes
are used. This method is expensive due to the high cost of enzymes but
it also offers high extraction efficiency as shown in Table 10 [234].

Table 10
Comparison for different methods of oil extraction from microalgae.

Methods Types Advantages Disadvantages Ref.

Physical extraction Pressing, Osmotic shock Ultrasonication, Simple and easy, High efficiency in cell Energy incentive, time consuming for drying, [159,221,228]
Bead milling, Microwave, Homogenization lysis, low fouling required cooling system due to high
temperature
Chemical solvent Soxhlet, FAME recovery increase by utilizing of High toxicity for environment, another [39,197,237]
extraction Bligh and dryer extraction polar and non-polar solvents, effective method also required to complete the
extraction
Biological extraction Enzymatic polysaccharide, Do not interfere with FA, high efficiency, Expensive [208,228]
Protein degradation
Supercritical fluid Supercritical CO2 extraction Green technology with low toxicity, No High operational cost due to energy incentive [159,236]
extraction chemical solvents, can be worked in wet
biomass

11
T. Fazal et al. Renewable and Sustainable Energy Reviews xxx (xxxx) xxx–xxx

Table 11 Chlorella vulgaris might produce almost 13.62 t/ha.year biodiesel [240].
Biodiesel production of different feedstock [42,78,171]. Besides production, biodiesel has several environmental advantages e.g.
it emits 78% less CO2, 98% less sulfur and 50% of particulate matter
Source Crop Name Oil yield Biodiesel productivity
(L ha−1) (kg biodiesel/ ha-year) emissions than other feedstocks [167]. On the other side, microalgal
biodiesel inherits similar characteristics to the standard biodiesel as
First generation Corn 172 152 shown in Table 12.
Soybean 446 562
Several studies have been carried out to produce biofuel from mi-
Canola 1190 862
Sunflower 1070 946 croalgae due to its rapid growth, high capacity for lipid accumulation
Palm Oil 5366 4747 (upto 1–85%), and ability to be cultivated in non-arable land (raceway,
Second Jatropha 741 656 photobioreactors etc) [37,235,241]. Besides several advantages, algal
generation Castor 1307 1156 biodiesel is not sufficiently cost competitive with conventional diesel
Third generation Microalgae (wet 58,700 51,927
[242]. In transesterification, the fatty acid profile for the FAMEs can
biomass)
Microalgae (Dry 136,900 121,104 obtained from Chlorella sp., Scenedesmus sp, Chlorella protothecoides, and
biomass) Chlorella vulgaris having unsaturated fatty acids as shown in Table 13
[176,187,243,244].

2.5.4. Supercritical fluid method
In supercritical fluid extraction, CO2 disrupts cell at high pressure
and temperature [36]. The extracted fatty acids (86%) in a SCCO2 was
obtained from green macroalgae [227]. This process offers better mass
transfer rate due to better diffusion of CO2 into microalgae cells. The oil
2.6.1. Transesterification
The extracted lipids are converted into biodiesel employing trans-
esterification process. Lipids (TAG and FFA) react with an alcohol
(methanol, ethanol, propanol, butanol and amyl alcohol) [249,250] to
form fatty acid methyl esters (FAME) and glycerol as shown in Eq. (7)
[94].

(7)

is extracted on depressurizing of CO2 [235]. This method is time effi-
cient because it requires 80 min than Soxhlet hexane extraction which
requires 330 min [159,236]. CO2 supercritical fluid extraction is a
greener approach because it causes neither toxicity nor thermal de-
gradation to extracts. This method yields non-contaminated residual
biomass, which can be easily used for animal feed, fertilizer or com-
posts and as a feedstock for anaerobic digestion. However, this process
requires high capital cost, high pressure equipment and operations, CO2
separation and compression after every oil extraction run [236].
2.6. Biodiesel production
Microalgae can be used to treat textile wastewater and other bior-
efinery applications such as human food, animal feed, cosmetic pro-
ducts, pharmaceutical products, fertilizers and biofuels [238,239]. Mi-
croalgae based biofuels can be divided into four categories based on
conversion technologies (thermo-chemical conversion, biochemical
conversion, transesterification and photosynthetic microbial fuel cell)
as shown in Fig. 5. However, microalgal biodiesel is the most important
option because it possesses similar physical and chemical character-
istics to petroleum as shown in Table 12. Biodiesel is a clean renewable
source of energy because it can be used in any compression ignition
engine without the need for modification [48]. Biodiesel is a mixture of
long chain fatty acid methyl esters (FAME) and is obtained by the
transesterification of lipids (Fig. 6). Biodiesel can also be produced from
nontoxic and biological resources such as vegetable oils, animal fats,
used cooking oil and algal oils. Microalgae do not need much cultiva-
tion land as compared to vegetable plants; however, it proves a better
feedstock for biodiesel as shown in Table 11. Recent study revealed that
Generally, methanol or ethanol is used, but methanol is preferred
due to low cost and physic-chemical advantages (polar and shortest
chain alcohol). Transesterification reaction is reversible therefore; ex-
cess supply of alcohol is used to ensure the forward reaction [251].
Biodiesel is produced by mixing an alcohol with TAG in a 3:1 M ratio.
Glycerol settle down at the bottom due to high density whereas FAME
remains on top [245]. Biodiesel yield can be obtained about 80% of the
volume of extracted algal oil [221].
The transesterification reaction is carried out in several steps. TAG
reacts with alcohol and get converted stepwise into diglycerides (DG)
which further converted into monoglycerides (MG). Finally, mono-
glycerides liberate one mol of FAME (biodiesel) at each step and gly-
cerol at the end as shown in the following Eq. (8) [252].
TG+R’OH ↔ DG+R’COOR1
DG+R’OH ↔ MG+R’COOR2
MG +R’OH ↔ Glycerol+R’COOR 3 (8)
Where, the R1, R2 and R3 are long chain hydrocarbons, also known as
fatty acid chains [48]. Normally six main types of fatty acid chains are
present in triglycerides of several microalgae species such palmitic,
palmitoleic, stearic, oleic, linoleic and linolenic acids. Table 13 presents
fatty acids profile of different microalgal species.
Biodiesel production depends on several factors such as reaction
temperature, reaction time, conc. of alcohol and catalyst. Generally,
chemical (acids, bases) or biological (enzymes) catalysts are used in
transesterification reaction [47]. Mostly, base catalysts are used for
biodiesel production such as NaOH, KOH, Al2O3, BaO, SrO, Cao, zeo-
loites, carbonates and corresponding sodium and potassium alkoxides

12
T. Fazal et al. Renewable and Sustainable Energy Reviews xxx (xxxx) xxx–xxx

Microalgae

Microbial Fuel Biochemical Thermochemical

Transesterication
Cell Conversion conversion

Acid/Base Supercritical
Catalysis Fluid

Anaerobic
Fermentation
Digestion

Photo-biological
Hydrogen
production

Biomethane/
Bioelectricity Biodiesel Biohydrogen Bioethanol
Biogas

Direct
Pyrolysis Liquefaction Gasification
Combustion

Electricity Bio-oil/charcoal Bio-oil Syngas

Fig. 5. Microalgae biomass conversion processes into biofuels [245].

such as sodium methoxide (Na-OCH3), sodium ethoxide (Na-OC2H5), [252,253]. Alkali-catalyzed transesterification is much faster (4000
sodium propoxide (Na-OC3H7) and sodium butoxide (Na-OC4H9) times) compared to acid catalysts [251]. The reaction is very fast but
[47,187,242]. Sulfuric acid, sulfonic acid and hydrochloric acid are high free fatty acids (FFA) content (>2%) can lead the reaction to sa-
used as acid catalysts and lipase can be used as enzymatic biocatalysts ponification [159]. The reaction takes place in four steps as given in Eq.
(9) [251].
First step:
Table 12
Comparison of characteristics of microalgal Biodiesel, conventional diesel fuel and ASTM NaOH +R′’OH ↔Na R′O- + H2O
biodiesel standard [129,246].
NaR’O- ↔R′O- + Na+
Properties ASTM biodiesel Diesel fuel Microalgal
biodiesel Second step:
2 −1
Viscosity (mm s , cSt 3.5 − 5.0 1.9 − 4.1 5.2 ROCOR1 + R’O- ↔ R1COROR’O
at 40 °C
Density (kg L−1) 0.84–0.90 0.838 0.864 Third step:
Solidifying point (°C) – −50 to 10 −12
Flash point (°C) Min 100 75 115
R1COROR’O- + R′OH↔ R1COROR’HO- + OR’
Cold filter plugging Summer max 0 −3.0 to −11
point (°C) Winter max < −6.7
Final step:
−15
R1COROR’HO- ↔ R1COOR’ + HOR (9)
Acid value (mg KOH Max 0.5 Max 0.5 0.373
g−1)
For an alkaline transesterification, triglycerides and alcohol must be
Heating value (MJ – 40 – 45 41
Kg−1)
substantially anhydrous [116] because water produced during trans-
H/C ratio – 1.81 1.81 esterification, leads the reaction to saponification. The formation of

13
T. Fazal et al. Renewable and Sustainable Energy Reviews xxx (xxxx) xxx–xxx

Fig. 6. Flow diagram of enzyme transesterification

Lipase + MeOH for FAME production [249].

Upper Phase

Extracted Oil Transesterification Separation FAME

Lower Phase

Purification of
glycerol

soap reduces the yield of esters, hinders the separation of ester and
glycerol, and causes catalyst loss [251]. The typical hydrolysis and soap
formation reactions are given in Eqs. 10 and 11 [48].
Hydrolysis
TAG+H2O→DG+ R1-COOH
Soap formation
R1-COOH + NaOH→R1COONa + H2O
FFA Sodium hydroxide Soap water
Acid catalyzed transesterification is effective for the conversion of
high content of FFA (>2%) into esters [39,47,159]. Acid catalyzed
transesterification is very slow and achieving on high temperature
(100 °C) and pressure, and thus, becomes expensive [165]. Reaction is
given in Eq. (1)2 [246].
R1—COOH + ROH→R—O—CO—R1+ H2O (12)
(10)
FFA Alcohol FAME water
Chemical catalysis is energy intensive, requires removal of catalyst
from the products, causes risk of saponification and makes glycerol
(11)
recovery difficult [47].
Enzymes can be used for transesterification because it exhibits good
tolerance to FFA content of the lipids as shown in Fig. 6. Lipase
transesterification is carried out at 35–45 °C which ensures no foam

Table 13
Composition of fatty acids in microalgae feedstock.

Fatty acids Microalgae feedstock

Fatty acids Comp Mol wt. Sp Cv Ch Sc Nan Bb Bs Cp Cs

[247] [186] [125] [161] [247] [161] [187] [248] [141]

Myrisitic C14:0 242 0.34 – 1.61 1.06 7.16 0.73 – 0.68 –

Palmitic C16:0 270 40.1 24.0 50.7 52.07 23.35 7.17 – 8.04 19.2
Palmitoleic C16:1, n=7 268 9.19 2.10 0.28 – 26.87 4.8 0.25 –
Steararic C18:0 298 1.18 1.32 11.54 7.48 0.45 1.59 4.3 3.17 –
Oleic C18:1, n=9 296 5.43 24.77 7.82 21.46 13.20 77.88 55.7 70.95 11.2
Linoleic C18:2, n=6 17.8 47.8 3.93 4.60 1.21 5.16 34.2 14.99 18.0
Linolenic C18:3 0.82 2.83 – 5.34 0.2 1.15 10.5
Eicosenoic C20:0 326 0.06 – – – – – – 0.09 –
Total saturated 41.68 25.32 63.9 78.61 14.31 9.49 4.3 11.98 19.2
Total unsaturated 51.9 74.67 12.8 21.39 30.96 88.38 94.9 87.34 39.7
Total FA 93.6 99.9 76.7 100 45.27 97.87 99.2 99.32 58.9

Note* Spirulina maxima (Sp), Chlorella vulgaris (Cv), Chlamydomonas sp. (Ch), Scenedesmus obliquus (Sc), Nannochloropsos sp (Nan), Botryoccus braunii (Bb), Botryococcus sp. (Bs),
Chlorella protothecoides (Cp), Chlorella sp. (Cs).

Table 14
Comparison of typical transesterification [48,116,246].

Type of Transesterification Advantages Disadvantages Limitations

Acidic Transesterification
Alkali Transesterification
Enzymatic Transesterification
Direct transesterification
Useful for high FFA content in lipids,
avoid soap formation, recoverable
4000 times faster, high efficiency
(90%)
Fast, easy recovery of esters, lower
alcohol to oil ratio required,
Good tolerance to the FFA, no pollution
to nature, no saponification
Elimination of oil extraction step,
reduce time and cost
Time consuming, removal of acid need large
amount of water,
Equipment corrosion, high cost
Saponification, difficult recovery of glycerol, loss
of catalyst
Recovery of glycerol not easy due to complex
formation of immobilized lipase with it,
denaturation
High chemical demand, high temperature
Works at high temperature. Long reaction times,
High alcohol and catalyst required for avoiding
soap formation, useful for low FFA contents
(<2%)
Expensive due to enzyme cost at larger scale
Co-solvent need to maintain the polarity of
reaction

14
T. Fazal et al.
formation catalytic activity [116]. This process is advantageous because
it requires less energy, low molar ratio of alcohol to oil and easy re-
covery of esters [254]. The immobilized lipase surface adheres with
glycerol and it diminishes enzymatic activity during transesterification
reaction. Cost of lipase enzyme and the recovery of glycerol from im-
mobilized lipase surface limit this method for commercial scale [246].
Proper immobilization of the enzyme and multiple enzymes may pro-
vide more choices for enzymatic transesterification in the future.
Transesterification products are mixture of esters, glycerol, alcohol,
catalyst and tri-, di- and monoglycerides. Pure esters cannot be ob-
tained due to the presence of impurities in the esters, such as di- and
monoglycerides. Glycerol is a by-product which needs to be recovered
by gravitational settling or centrifuging.
In situ (direct) transesterification, biodiesel can be produced by
combing extraction and transesterification processes into one-step. The
harvested algal cells are directly transesterified into fatty acid esters,
eliminating oil extraction step with the addition of alcohol and catalyst
[199]. This single step transesterification can save time and cost resulting
in high algae conversion to FAME yield (98%) [159]. Table 14 compares
different transesterification processes. Chemical transesterification is
most commonly employed process for algal biodiesel production.
3. Future prospects and conclusion
Several studies have already established that microalgae cultivation
with a single objective i.e. for biodiesel production will not be sus-
tainable option because; the cost of nutrients and other supplies cannot
make this biodiesel economical and feasible against commercial bio-
diesel. Thus, it is important to reorient future research and development
focus on microalgae biodiesel production [78]. Recent studies have
feasibly redefined microalgae double role where it can be simulta-
neously employed to remediate industrial wastewater (e.g. TWW) and
then to be used for biodiesel production. This double role can be further
investigated in various dimensions to optimize wastewater treatment
and biodiesel production. There are several following avenues, which
can be investigated:
• Microalgal cultivation in TWW is mainly carried out at lab scale
however pilot scale studies are still scarce. There are several chal-
lenges microalgae may face at pilot scale cultivation in TWW such as
microbial contamination, variable composition of wastewater (dye
and nutrient basis), inconsistent biomass productivity, variable lipid
yield etc. [41]. These factors are required to be properly investigated.
• TWW composition varies from batch. Thus, robust microalgal spe-
cies are required to be cultivated for better productivity and lipid
yield. Therefore, new species should be screened and isolated from
real TWW and optimized for simultaneous bioremediation and
biodiesel production [41,255].
•
The phosphorus concentration influences the lipid content in mi-
croalgae. An inaccurate N/P ratio may limit lipid accumulation
during cultivation [255]. TWW composition may face variable N/P
ratio. In this regard, microalgae isolated from real TWW should be
optimized for an appropriate N/P in lab scale experiments. Later,
isolated microalgae should be cultivated in a mixed wastewater
stream with optimum N/P ratio [256] for better lipid yield.
• Bacteria are already present in textile wastewater. There is a likely
growth competition between bacteria and microalgae. At the same
time, this bacteria-algal consortium may synergistically speed up
wastewater treatment [100]. Thus, the role of this consortium needs
to be investigated on growth and wastewater treatment scales using
real and synthetic wastewater.
•
Different microalgae (Chlorella pyrenoidosa, Chlorella vulgaris and
Oscillatoria tenuis) have biodegraded around 30 azo dyes into simpler
compounds [257]. Besides biodegradation of dyes, microalgae also
remove dyes from wastewater via adsorption process. Thus, the role
of microalgae in both the phenomena should be investigated.
15
Renewable and Sustainable Energy Reviews xxx (xxxx) xxx–xxx
Moreover, spent microalgae can also be used to produce activated
carbon and/or biochar, which may remove high concentrations of
pollutants.
Acknowledgement
This work has been carried out under CIIT, Pakistan and IPE, China
research collaboration. MSU Rehman, F Rehman and T Fazal are greatly
thankful to the Higher Education Commission (HEC), Pakistan for pro-
viding generous financial assistance under NRPU Project No. 4547. MSU
Rehman (2016PT012) and AU Khan (2017VEA0029) acknowledge fel-
lowship under President's International Fellowship Initiative (PIFI),
Chinese Academy of Sciences, Beijing, China. Authors also acknowledge
National Natural Science Foundation of China (21176238, 21576266), the
Major Research plan of the National Natural Science Foundation of China
(91534107), Wanjiang Scholar Program and Start Fund for Biochemical
Engineering Center from Anhui University of Technology, China.
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