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Scenedesmus-based Treatment of Nitrogen and

Phosphorus from Effluent of Anaerobic
Digester and Bio-oil Production
ARTICLE in BIORESOURCE TECHNOLOGY NOVEMBER 2015
Impact Factor: 5.04 DOI: 10.1016/j.biortech.2015.07.091

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Korea Advanced Institute of Science and Te

University of Hawaii

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Bioresource Technology 196 (2015) 235240

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Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Scenedesmus-based treatment of nitrogen and phosphorus from efuent

of anaerobic digester and bio-oil production
Ga-Yeong Kim a, Yeo-Myeong Yun b, Hang-Sik Shin a, Hee-Sik Kim c, Jong-In Han a,
a

Department of Civil and Environmental Engineering, KAIST, 373-1 Guseong-dong, Yuseong-gu, Daejeon 305-701, Republic of Korea
Forestry and Natural Resource Management, College of Agriculture, University of Hawaii at Hilo, 200 W. Kawili St., Hilo, HI 96720, United States
c
Sustainable Bioresource Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 305-806, Republic of Korea
b

h i g h l i g h t s
 Synergetic coupling of wastewater treatment and biodiesel production was assessed.
 The efuent of anaerobic digester was treated by Scenedesmus sp.
 The nitrogen and phosphorus were effectively removed from the efuent.
 High biomass productivity and FAME productivity were obtained at the same time.

a r t i c l e

i n f o

Article history:
Received 1 June 2015
Received in revised form 22 July 2015
Accepted 23 July 2015
Available online 29 July 2015
Keywords:
Anaerobic digestion
Nitrogen removal
Phosphorus removal
Biodiesel
Scenedesmus sp.

a b s t r a c t
In this study, a microalgae-based technology was employed to treat wastewater and produce biodiesel at
the same time. A local isolate Scenedesmus sp. was found to be a well suited species, particularly for an
efuent from anaerobic digester (AD) containing low carbon but high nutrients (NH3-N = 273 mg L 1,
total P = 58.75 mg L 1). This algae-based treatment was quite effective: nutrient removal efciencies
were over 99.19% for nitrogen and 98.01% for phosphorus. Regarding the biodiesel production, FAME contents of Scenedesmus sp. were found to be relatively low (8.74% (w/w)), but overall FAME productivity
was comparatively high (0.03 g L 1 d 1) due to its high biomass productivity (0.37 g L 1 d 1). FAMEs
were satisfactory to the several standards for the biodiesel quality. The Scenedesmus-based technology
may serve as a promising option for the treatment of nutrient-rich wastewater and especially so for
the AD efuent.
2015 Elsevier Ltd. All rights reserved.

1. Introduction
The microalgae-based wastewater treatment has attracted
increasing attention due to its eco-friendly nature: it can consume
far less energy, with carbon emission greatly reduced, and also lead
to biofuel production.
Wastewater treatment, though indispensable, is an
energy-guzzling process. In the USA, up to 4% of electricity is consumed for the water and wastewater treatment (Daw et al., 2012).
This becomes more problematic when a tertiary treatment process,
such as biological nutrient removal (BNR), is operated in addition
to the conventional activated sludge facility (ASF). ASF and
ASF/BNR are known to consume 340660 kWh/mL (million liters
of wastewater) and 7401000 kWh/mL, respectively. The
microalgae-based treatment, on the other hand, requires far less
Corresponding author. Tel.: +82 42 350 3629; fax: +82 42 350 3610.
E-mail address: hanj2@kaist.ac.kr (J.-I. Han).
http://dx.doi.org/10.1016/j.biortech.2015.07.091
0960-8524/ 2015 Elsevier Ltd. All rights reserved.

energy of approximately 230 kWh/mL (Woertz et al., 2009).

Another distinctive advantage of the microalgae-based system is
its ability to contribute to carbon mitigation: 660 kg of CO2 can
be reduced per mL of wastewater in the microalgae-based facility
while 220 kg and 550 kg of CO2 are emitted in the ASF and
ASF/BNR, respectively (Woertz et al., 2009). Besides, resulting biomass of the algae-based treatment, specically lipids, can be transformed into biodiesel, with which the paradigm of wastewater
treatment
can
be
shifted
from
energy-consumer
to
energy-producer. Not only that, it can also play a positive role in
producing microalgae-derived biodiesel, an ever-promising green
transportation fuel, in such a way that wastewater and its treatment facility can serve as culture medium and infrastructure for
the microalgae biomass production.
Thus far, there have been several attempts aimed to treat various types of wastewater, such as municipal wastewater, dairy
manure, and soybean processing wastewater, with microalgae
(Hongyang et al., 2011; Ji et al., 2014; Wilkie and Mulbry, 2002).

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G.-Y. Kim et al. / Bioresource Technology 196 (2015) 235240

In this study, we particularly focused on the efuent of anaerobic

digestion (AD) system. This is mainly because it has low organic
but high nitrogen and phosphorus contents, all of which are advantageous for the algae cultivation in particular and algae-based
technology in general; the rich nitrogen and phosphorus in the
AD efuent can serve as good nutrients sources for the microalgae
cultivation (Noike et al., 2004; Park et al., 2010). The common tertiary treatment technologies, to the contrary, require high C/N ratio
for optimal treatment; for example, A2O (Anaerobicanoxicoxic)
system requires about 57 of C/N ratio for the effective nutrients
removal since organic carbon sources are essential for the growth
of denitrier and polyphosphate-accumulating organisms (PAO)
(Xiaolian et al., 2006). This condition is rather strict and thus the
addition of external organic carbons is often needed. The microalgae, on the other hand, are in no need of such organic substrates, as
they grow on inorganic carbon source, carbon dioxide, and it can be
provided even from the biogas produced in the AD. Another unique
feature of the AD efuent used in this study was its comparatively
high initial contents of nitrogen and phosphorus (Table 1)
(Abou-Shanab et al., 2013; Aravantinou et al., 2013; Cabanelas
et al., 2013; Komolafe et al., 2014; Zhang et al., 2013).
The primary aim of this study was therefore to treat this
nutrient-rich wastewater from the AD to a level that meets the discharge water quality standards, and at the same time to produce
biodiesel, which was different from most of previous studies dealing with the AD efuent focused only on the treatment (Bjornsson
et al., 2013; Wang et al., 2014). All of these were attempted by
means of selecting and employing suitable microalgae species that
are active under high nutrients environments. In addition, lipid
productivity, together with the suitability of the resulting lipid as
a transportation fuel, were also assessed.

2. Methods
2.1. The efuent of anaerobic digestion system
An up-ow anaerobic sludge blanket (UASB) reactor system was
used for AD, with a seed taken from an AD at a local wastewater
treatment plant in Daejeon, Korea. Based on the previous study
(Jung et al., 2013), the UASB system was operated with a synthetic
wastewater, which had the following composition: glucose at a
concentration of 10,000 mg COD/L, NH4Cl, KH2PO4, FeCl2
4H2O for
COD:N:P:Fe ratio of 100:5:1:0.33, and trace elements (in mg L 1):
MgCl2
6H2O 200; CaCl2
2H2O 150; Na2MoO4
4H2O 0.02; H3BO3
0.10; MnCl2
4H2O 1.0; ZnCl2 0.10; CuCl2 0.06; NiCl2
6H2O 0.10;
CoCl2
2H2O 1.0; and Na2SeO3 0.10. Temperature and pH were continuously maintained as 37 C and 7 0.2, respectively. The efuent from this AD system was used for the subsequent algae

Table 1
Nitrogen and phosphorus contents of various wastewaters.
Wastewater

Nitrogen
(mg L 1)

Phosphorus
(mg L 1)

N/P
ratio

References

Piggery
wastewater
Facultative
lagoon
Primary efuent

56

13.5

4.2

42.3

35.4

1.2

27.7

1.6

17.3

Secondary
efuent
Domestic
wastewater
Anaerobic
digester
efuent

270330

68

52

7.1

33.8
55
7.3

273

58.8

4.6

Abou-Shanab
et al. (2013)
Komolafe et al.
(2014)
Zhang et al.
(2013)
Aravantinou
et al. (2013)
Cabanelas et al.
(2013)
In this study

cultivation after centrifugation at 3000 rpm for 5 min and

autoclave.
2.2. Microalgae strain and culture conditions
Scenedesmus sp., was isolated from a local wastewater treatment plant in Daejeon, Korea. It was maintained in BG11 medium,
which served as a control as well (Feng et al., 2011): NaNO3
(17.6 mM);
K2HPO4
(0.22 mM);
MgSO4
7H2O
(0.3 mM);
CaCl2
2H2O (0.2 mM); Citric Acid
H2O (0.03 mM); Ferric
Ammonium Citrate (0.02 mM); Na2EDTA
2H2O (0.002 mM);
Na2CO3 (0.18 mM); H3BO3 (46 lM); MnCl2
4H2O (9 lM);
ZnSO4
7H2O (0.77 lM); Na2MoO4
2H2O (1.6 lM); CuSO4
5H2O
(0.3 lM); and Co(NO3)2
6H2O (0.17 lM). A seed culture was prepared in a 250 mL bafed culture ask containing 100 mL of the
BG11 medium with continuous light of 140 lmol photons m 2 s 1
at 27 C and 150 rpm. The seed was added with 10% volumetric
concentration to a 250 mL bafed culture ask containing
200 mL of the autoclaved AD efuent, and cultivated for 23 days,
with operational conditions of constant shaking at 150 rpm, constant temperature at 27 C, continuous light with 140 lmol photons m 2 s 1, and 2% of CO2 air supplementation with 0.2 vvm.
Experiment were conducted with triplicate.
To shorten the length of cultivation, an additional experiment
was conducted with initial microalgae concentration increased.
For that, the cell culture grown up to 23 days was used as an inoculum, with all other conditions remained the same.
2.3. Analytical methods
The growth of microalgae was monitored by measuring optical
density and dry cell weight of microalgae. Optical density was
measured at 660 nm with UVVis spectrophotometer (DR 5000,
HACH) (Au et al., 2011), and the dry cell weight was obtained by
Standard Methods (APHA et al., 1999). The chemical oxygen
demand (COD) and the concentration of total phosphate (TP) and
ammoniacal nitrogen (NH3-N) were determined by water test kits
(Humas, HS-CODCr-MR, HS-TP-L, HS-NH3 (NW)-H) with UVVis
spectrophotometer (DR 5000, HACH). Also, the concentration of
organic acids was analyzed by a HPLC system (Finnigan Spectra
SYSTEM LC, Thermo Electron CO.) with 4 mM H2SO4 as mobile
phase. For the concentration of ions, inductively coupled plasma
optical emission spectroscopy (Agilent, USA) was used.
FAMEs (Fatty acid methyl esters) were also analyzed according
to the modied Folch method (Folch et al., 1957). A microalgae
sample was centrifuged at 2000 rpm for 10 min and washed with
distilled water once. This re-centrifuged cells were freeze-dried
at 52 C for 4 days. Afterwards, cells were powdered, and a solvent mixture of chloroform: methanol (2 mL, 2:1, v/v) was added
to the 10 mg of cells. After vortexing them vigorously for 20 min,
FAMEs were nally formed by adding 1 mL of methanol and
300 lL of H2SO4 at 100 C for 20 min. After being cooled down,
1 mL of distilled water was added to the sample, which was then
vortexed for 5 min and centrifuged at 4000 rpm for 10 min. The
lower layer including the organic solvent was analyzed with gas
chromatography (HP 5890, Agilent, USA) with a ame ionized
detector (FID) and INNOWAX capillary column (Agilent, USA).
3. Results and discussions
3.1. Characterization of the efuent from anaerobic digester
Basic characteristics of the UASB efuent are summarized in
Table 2. As with the typical AD process, COD was substantially
reduced from 10,000 mg L 1 to 660 mg L 1. The removal of

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G.-Y. Kim et al. / Bioresource Technology 196 (2015) 235240

Table 2
Characteristics of the inuent and efuent from an UASB reactor.
Parameters

Inlet concentration
(mg L 1)

Chemical oxygen demand

(COD)
Volatile fatty acid (formic
acid)
Acetate
Propionate
NH3-N
TP
Ions
Ca2+
K+
Mg2+
Na+
S2
Si4+
Al3+
Co2+
Cu2+
Fe2+
Mn2+
Ni2+
Li+
Zn2+

10,000.00

660.00

224.58

500.00
100.00

74.26
150.32
273.00
58.75

40.82
125.80
23.65
0.03
0.00
0.00
0.00
0.17
0.03
33.00
0.28
0.02
0.00
0.05

32.54
126.80
10.46
1049.00
361.80
42.13
0.25
0.03
0.03
1.16
0.02
0.00
0.07
0.01

Outlet concentration
(mg L 1)

nitrogen and phosphorus, on the other hand, was quite limited:

about half of them still remained untreated, with the nal concentrations of 273 mg L 1 for nitrogen and 58.75 mg L 1 for phosphorus. These were much higher than the nutrients included in the
typical domestic wastewater, and this noticeable inefciency warranted a dedicated tertiary treatment. Among several options, a
microalgae-based technology was adopted to treat the remaining
nutrients in this AD efuent.
For microalgae to grow actively in the wastewater, the ratio of
nitrogen and phosphorus is important. Assuming algal biomass
has a chemical formula of C106H181O45N16P, 7.2 of N/P mass ratio
would be theoretically optimal, but in reality, it has a wider range
of from 4 to 45 depending on the species (Craggs et al., 2011;
Klausmeier et al., 2004). In particular, the optimal N/P ratio of
Scenedesmus sp. was known to be 530 depending on the ecological
condition (Choi and Lee, 2014). Therefore, the N/P ratio of 4.6 of the
AD efuent used in this study, though on the extreme end, fell
within the range, and may well support the algal growth so long
as an appropriate species is chosen.
In addition, minerals other than nitrogen and phosphorus were
also high, signicantly more than in the BG11 medium. This is the
reason that salt-resistant species were needed to use and thus subsequently sought.

Fig. 1. The growth and nutrients removal of Scenedesmus sp. in the efuent from
anaerobic digester: (a) growth; and (b) removal of nitrogen and phosphorus.

Table 3
Summary of Scenedesmus sp. cultivation in the AD efuent.
Parameters

Initial biomass concentration (g L 1)

Final biomass concentration (g L 1)
Biomass productivity (g L 1 d 1)
Nitrogen removal efciency (%)*
Phosphorus removal efciency (%)
FAME contents (w/w%)
FAME productivity (g L 1 d 1)

3.2. Growth and nutrients removal of microalgae

A local isolate Scenedesmus sp., which had been originally isolated from a wastewater, was found to grow well in the AD efuent, which has low N/P ratio and rather high salts, with the
biomass productivity of 0.37 g L 1 d 1 (Fig. 1(a) and Table 3).
These values, which were in fact quite high, were comparable to
other previous studies (Table 4) (Bjornsson et al., 2013;
Cabanelas et al., 2013; Cho et al., 2011; Ji et al., 2014; Wang
et al., 2014; Xue et al., 2010). It was likely attributable to the high
initial
concentrations
of
nitrogen
and
phosphorus
(NH3-N = 273 mg L 1, TP = 58.75 mg L 1).
The intended removal of nutrients was also successfully
achieved: the nal concentration of nitrogen was less than
2 mg L 1 (over 99.19% of removal efciency) and that of phosphorus was 1.05 mg L 1 (removal efciency of 98.01%) (Fig. 1(b) and

Scenedesmus sp.

Lower limit of nitrogen detection was 2 mg L

Efuent

BG 11

0.09
8.55
0.37
>99.19
98.01
8.74
0.03

0.12
9.55
0.41

11.72
0.05

Table 3). Both levels were satisfactory to the discharge water quality standards of Korea, which are 20 mg L 1 for nitrogen and 0.2
2 mg L 1 for phosphorus.
The N/P ratio balance is critical for the algae growth and thus
nutrients removal, and is known to vary greatly depending on species (Wang et al., 2014). The selected local isolate Scenedesmus sp.
appeared to be well tted to the AD efuent in this study.

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G.-Y. Kim et al. / Bioresource Technology 196 (2015) 235240

Table 4
Biomass and FAME productivities of various previous studies.
Wastewater

Microalgae

Biomass productivity
(g L 1 d 1)

FAME productivity
(g L 1 d 1)

References

Secondary treated wastewater

Concentrated municipal wastewater + 3% mine
wastewater
Domestic wastewater + glycerol addition

Chlorella sp.
Micractinium reisseri

0.07
0.05

0.02
0.01

Cho et al. (2011)

Ji et al. (2014)

0.12
0.28
0.32

0.02*
0.04*
0.04

Cabanelas et al.
(2013)
Xue et al. (2010)

0.07
0.06
0.07

Wang et al. (2014)

AD efuent (diluted)

Chlorella vulgaris
Botryococcus terribilis
Spirulina platensis + Rhodotorula
glutinis mix
Chlorella sp.
Micractinium sp.
Scenedesmus sp.

AD efuent

Scenedesmus sp.

0.37

0.03

Bjornsson et al.
(2013)
This study

Monosodium glutamate wastewater

Mixture of AD concentrate and primary efuent

Lipid productivity.

Scenedesmus sp. could effectively metabolize both of nitrogen and

phosphorus without suppressed assimilation of any nutrients.
To shorten the treatment time, an increased amount of inoculum was added (Fig. 2). In this way, one batch cultivation was completed within 13 days, which was considerably shorter than the
rst treatment period of 23 days; and the biomass productivity
was increased from 0.37 g L 1 d 1 to 0.74 g L 1 d 1. The lag time
was literally eliminated likely because of the use of the

already-adapted Scenedesmus sp. cells. It was observed that the

nutrients were removed as the rst-order reaction from the
beginning.
3.3. FAME productivity and composition
Lipids were extracted from the cultivated microalgae biomass,
and converted into FAME, which is the main component of biodiesel. The FAME contents of Scenedesmus sp. were found to be 8.74%
(w/w) that was unacceptably low. It is likely attributable to the
high initial concentrations of nitrogen and phosphorus. It is known
that the microalgae accumulate the lipid with the depletion of
nitrogen and phosphorus (Cho et al., 2011); therefore, a
two-stage approach, in which biomass production and lipid accumulation take place in a separate and consecutive way, may be a
cultivation method of choice to attain the aimed lipid productivity.
In this study, FAME productivity of 0.03 g L 1 d 1, however, was
comparable to other studies due to its relatively high biomass productivity, 0.37 g L 1 d 1 (Table 4).
The resulting lipid was also assessed in terms of its quality as
well as quantity for the biodiesel production. Among various
lengths of fatty acids found in microalgae, C8C24, C16C18 are
regarded as the best for the fuel purpose (Tang et al., 2011).
Interestingly, 76.68% (w/w) of C16C18 was obtained in this study
(Table 5). In addition to the length of carbon chain, degree of saturation is also important to be used as fuel. In this respect,
mono-unsaturated fatty acids (MUFAs) are considered as the best
for the biodiesel purpose. With high contents of MUFAs in the
extracted FAME, reasonable balance of fuel properties such as

Table 5
FAME (fatty acid methyl esters) prole and lipid quality assessment.

Fig. 2. Re-cultivation of Scenedesmus sp. with an increased amount of inoculum: (a)

recovery of growth; and (b) recovery of nutrients removal.

FAME type

Percentage of certain FAME amounts in the total

FAME(% w/w)
Scenedesmus sp.

C16:0 (Palmitic)
C16:1n9c
(Palmitoleic)
C18:0 (Stearic)
C18:1n9t (Oleic)
C18:2n6c (Linoleic)

25.51
1.86
19.46
10.23
17.49

C16C18
Other
Mono-unsaturated
Cetane number
Oxidation stability
Iodine value

76.68
25.45
12.09
58.16
9.33
47.65

FAME contents* (%)

8.74

FAME contents = FAME (mg)/dry cell weight of biomass (mg)  100.

G.-Y. Kim et al. / Bioresource Technology 196 (2015) 235240

ignition quality, combustion heat, cold lter plugging point (CFPP),

oxidation stability, viscosity and lubricity can be obtained (Knothe,
2008).
Other criteria for biodiesel, such as cetane number, oxidation
stability and iodine value were also calculated and compared with
the European Standard (EN 14214) describing the requirements of
biodiesel (Table 5); the criteria for cetane number, oxidation stability, and iodine value are >51, >6, and <120, respectively. Cetane
number is related to the ignition delay and combustion quality,
and the cetane number of Scenedesmus sp. oil was satisfactory to
the European Standard. Also, it was higher than the commercialized vegetable oils such as jatropha, soybean, and sunower oil
(Seo et al., 2014). The oxidation stability and iodine value, which
are related to the unsaturated fatty acids, were also satisfactory
due to the high contents of saturated fatty acids such as C16:0
and C18:0.
3.4. Potential economic benets of integrated algae-based process for
wastewater treatment
One distinctive advantage of the microalgae-based treatment
technology is its potential of producing diverse value products.
Since biodiesel production only, though imperative, less likely
makes the microalgae-based process economically competitive
(Likozar and Levec, 2014; otaric et al., 2012), other prot routes
termed microalgae biorenery must be sought: some exemplary
products include food additives, cosmetics, pharmaceuticals, animal feed, and industrial chemicals (Uggetti et al., 2014). In this
regard, a blueprint of the integrated process grounded up the AD
efuent was proposed (Table 6), and its feasibility was assessed
on the basis of the results of this study.
An AD was assumed to have a capacity of 200 m3 d 1, and be
operated for 300 days per year. With these assumptions,
22,200 kg y 1 of biomass productivity and 1800 kg y 1 of FAME
productivity can be obtained. Assuming 10% (w/w) of oil feedstock
is converted into glycerol, 180 kg y 1 of glycerol is produced (Yang
et al., 2012). Likewise, 6882 kg y 1 of biochar, 2220 kg y 1 of nitrogen fertilizer, and 222 kg y 1 of phosphorus fertilizer can be produced alongside (Chaiwong et al., 2012; Grobbelaar, 2004). This
avenue is particularly noteworthy, as nitrogen fertilizer production
is highly energy-consuming, and phosphorus is a depleting
resource.

Table 6
Estimates of productivities of other valuable materials: with the assumption of
anaerobic digestion capacities of 200 m3 d 1 and with 300 days per year.
Parameters

Scenedesmus sp.
1

Biomass productivity (kg y )

FAME productivity (kg y 1)
Biodiesel from FAME (kg y 1)a
Glycerol generated (kg y 1)b
Biochar (kg y 1)c
Fertilizer (N kg y 1)d
Fertilizer (P kg y 1)e
Methane (m3 L y 1)f
Hydrogen (m3 L y 1)g
a

22,200
1800
1800
180
6882
2220
222
8746.8
1021.2

Here estimated that all FAME are fully converted into the biodiesel.
Conversion rate of 0.1 kg of glycerol per kg of oil feedstock (Yang et al., 2012).
c
The biochar production yield is 31% (w/w) of microalgae biomass (Chaiwong
et al., 2012).
d
10% (w/w) of biomass for the nitrogen fertilizer and 1% (w/w) of biomass for the
phosphorus fertilizer (Grobbelaar, 2004).
e
10% (w/w) of biomass for the nitrogen fertilizer and 1% (w/w) of biomass for the
phosphorus fertilizer (Grobbelaar, 2004).
f
Here assumed that 394 L of methane and 46 L of hydrogen are produced per kg
of biomass (Yang et al., 2011).
g
Here assumed that 394 L of methane and 46 L of hydrogen are produced per kg
of biomass (Yang et al., 2011).
b

239

Moreover, by way of anaerobic digestion of algae residue,

8746.8 m3 L y 1 of methane and 1021.2 m3 L y 1 of hydrogen gas
can be yielded based on the assumption that 394 L of methane
and 46 L of hydrogen would be produced per kg of biomass
(Yang et al., 2011). Considering all of these, the following integration is possible: (1) microalgae is cultivated while treating the AD
efuent; (2) microalgae biomass is used for the biodiesel production; (3) the lipid-extracted algal residues are returned to AD; (4)
biogas is produced from AD, and energy is generated with the biogas; (5) this electricity is used to maintain the wastewater treatment facilities; as a result, (6) the overall treatment cost and
energy consumption of wastewater treatment plant can be
reduced.
4. Conclusions
This study demonstrated that high-strength efuent of anaerobic digester could be effectively treated by using microalgae,
Scenedesmus sp. This algae technology appeared to be particularly
suited for the wastewater with low C/N ratio but high nitrogen
and phosphorus, which has been troublesome with the conventional tertiary treatment. Not only that, its biodiesel productivity
was also comparable to other studies using different wastewater
sources. In order to realize the potential of this microalgae-based
technology, continuous and in-depth investigations on the cultivation particularly coupled with biorenery concept are warranted.
Acknowledgements
This research was supported by the Advanced Biomass R&D
Center (ABC) through the National Research Foundation of Korea
(NRF) funded by the Ministry of Science, ICT and Future Planning
(NRF-2011-0031348) and National Research Foundation of Korea
(NRF) grant funded by the Korea government Ministry of
Education,
Science
and
Technology
(MEST)
(NRF-2012M1A2A2026587).
References
Abou-Shanab, R.A.I., Ji, M.K., Kim, H.C., Paeng, K.J., Jeon, B.H., 2013. Microalgal
species growing on piggery wastewater as a valuable candidate for nutrient
removal and biodiesel production. J. Environ. Manage. 115, 257264.
APHA, AWWA, WEF, 1999. Standard Methods for the Examination of Water and
Wastewater, 20th ed. American Public Health Association.
Aravantinou, A.F., Theodorakopoulos, M.A., Manariotis, I.D., 2013. Selection of
microalgae for wastewater treatment and potential lipids production.
Bioresour. Technol. 147, 130134.
Au, S.H., Shih, S.C.C., Wheeler, A.R., 2011. Integrated microbioreactor for culture and
analysis of bacteria, algae and yeast. Biomed. Microdevices 13, 4150.
Bjornsson, W.J., Nicol, R.W., Dickinson, K.E., McGinn, P.J., 2013. Anaerobic digestates
are useful nutrient sources for microalgae cultivation: functional coupling of
energy and biomass production. J. Appl. Phycol. 25, 15231528.
Cabanelas, I.T.D., Arbib, Z., Chinalia, F.A., Souza, C.O., Perales, J.A., Almeida, P.F.,
Druzian, J.I., Nascimento, I.A., 2013. From waste to energy: microalgae
production in wastewater and glycerol. Appl. Energy 109, 283290.
Chaiwong, K., Kiatsiriroat, T., Vorayos, N., Thararax, C., 2012. Biochar production
from freshwater algae by slow pyrolysis. J. Sci. Technol. 6, 186195.
Cho, S., Luong, T.T., Lee, D., Oh, Y.K., Lee, T., 2011. Reuse of efuent water from a
municipal wastewater treatment plant in microalgae cultivation for biofuel
production. Bioresour. Technol. 102, 86398645.
Choi, H.J., Lee, S.M., 2014. Effect of the N/P ratio on biomass productivity and
nutrient removal from municipal wastewater. Bioprocess Biosyst. Eng. 38, 761
766.
Craggs, R.J., Heubeck, S., Lundquist, T.J., Benemann, J.R., 2011. Algal biofuels from
wastewater treatment high rate algal ponds. Water Sci. Technol. 63, 660665.
Daw, J., Hallett, K., DeWolfe, J., Venner, I., 2012. Energy Efciency Strategies for
Municipal Wastewater Treatment Facilities. NREL.
Feng, Y., Li, C., Zhang, D., 2011. Lipid production of Chlorella vulgaris cultured in
articial wastewater medium. Bioresour. Technol. 102, 101105.
Folch, J., Lees, M., Stanley, G.H.S., 1957. A simple method for the isolation and
purication of total lipids from animal tissues. J. Biol. Chem. 226, 497509.
Grobbelaar, J.U., 2004. Algal nutrition: mineral nutrition. In: Richmond, A. (Ed.),
Handbook of Microalgal Culture: Biotechnology and Applied Phycology.
Blackwell Publishing, Oxford, pp. 95115.

240

G.-Y. Kim et al. / Bioresource Technology 196 (2015) 235240

Hongyang, S., Yalei, Z., Chunmin, Z., Xuefei, Z., Jinpeng, L., 2011. Cultivation of
Chlorella pyrenoidosa in soybean processing wastewater. Bioresour. Technol.
102, 98849890.
Ji, M.K., Kabra, A.N., Salama, E.S., Roh, H.S., Kim, J.R., Lee, D.S., Jeon, B.H., 2014. Effect
of mine wastewater on nutrient removal and lipid production by a green
microalga Micratinium reisseri from concentrated municipal wastewater.
Bioresour. Technol. 157, 8490.
Jung, K.W., Cho, S.K., Yun, Y.M., Shin, H.S., Kim, D.H., 2013. Rapid formation of
hydrogen-producing granules in an up-ow anaerobic sludge blanket reactor
coupled with high-rate recirculation. Int. J. Hydrogen Energy 38, 90979103.
Klausmeier, C.A., Litchman, E., Daufresne, T., Levin, S.A., 2004. Optimal nitrogen-tophosphorus stoichiometry of phytoplankton. Nature 429, 171174.
Knothe, G., 2008. Designer biodiesel: optimizing fatty ester composition to
improve fuel properties. Energy Fuels 22, 13581364.
Komolafe, O., Velasquez Orta, S.B., Monje-Ramirez, I., Noguez, I.Y., Harvey, A.P., Orta
Ledesma, M.T., 2014. Biodiesel production from indigenous microalgae grown
in wastewater. Bioresour. Technol. 154, 297304.
Likozar, B., Levec, J., 2014. Effect of process conditions on equilibrium, reaction
kinetics and mass transfer for triglyceride transesterication to biodiesel:
experimental and modeling based on fatty acid composition. Fuel Process.
Technol. 122, 3041.
Noike, T., Goo, I.S., Matsumoto, H., Miyahara, T., 2004. Development of a new type of
anaerobic digestion process equipped with the function of nitrogen removal.
Water Sci. Technol. 49, 173179.
Park, J., Jin, H.-F., Lim, B.-R., Park, K.-Y., Lee, K., 2010. Ammonia removal from
anaerobic digestion efuent of livestock waste using green alga Scenedesmus sp.
Bioresour. Technol. 101, 86498657.
Seo, Y.H., Han, S., Han, J.-I., 2014. Economic biodiesel production using algal residue
as substrate of lipid producing yeast Cryptococcus curvatus. Renewable Energy
69, 473478.
otaric, M., Klinar, D., Bricelj, M., Golob, J., Berovic, M., Likozar, B., 2012. Growth,
lipid extraction and thermal degradation of the microalga Chlorella vulgaris.
New Biotechnol. 29, 325331.

Tang, D., Han, W., Li, P., Miao, X., Zhong, J., 2011. CO2 bioxation and fatty acid
composition of Scenedesmus obliquus and Chlorella pyrenoidosa in response to
different CO2 levels. Bioresour. Technol. 102, 30713076.
Uggetti, E., Sialve, B., Trably, E., Steyer, J.-P., 2014. Integrating microalgae production
with anaerobic digestion: a biorenery approach. Biofuels Bioprod. Bioren. 8,
516529.
Wang, M., Kuo-Dahab, W.C., Dolan, S., Park, C., 2014. Kinetics of nutrient removal
and expression of extracellular polymeric substances of the microalgae,
Chlorella sp. and Micractinium sp., in wastewater treatment. Bioresour.
Technol. 154, 131137.
Wilkie, A.C., Mulbry, W.W., 2002. Recovery of dairy manure nutrients by benthic
freshwater algae. Bioresour. Technol.
Woertz, I., Fulton, L., Lundquist, T., 2009. Nutrient removal & greenhouse gas
abatement with CO2 supplemented algal high rate ponds. Proc. Water Environ.
Fed. 2009, 79247936.
Xiaolian, W., Yongzhen, P., Shuying, W., Jie, F., Xuemei, C., 2006. Inuence of
wastewater composition on nitrogen and phosphorus removal and process
control in A2O process. Bioprocess Biosyst. Eng. 28, 397404.
Xue, F., Miao, J., Zhang, X., Tan, T., 2010. A new strategy for lipid production by mix
cultivation of Spirulina platensis and Rhodotorula glutinis. Appl. Biochem.
Biotechnol., 498503
Yang, F., Hanna, M.A., Sun, R., 2012. Value-added uses for crude glycerol a
byproduct of biodiesel production. Biotechnol. Biofuels 5, 13.
Yang, Z., Guo, R., Xu, X., Fan, X., Luo, S., 2011. Hydrogen and methane production
from lipid-extracted microalgal biomass residues. Int. J. Hydrogen Energy 36,
34653470.
Zhang, T.-Y., Wu, Y.-H., Zhu, S., Li, F., Hu, H.-Y., 2013. Isolation and heterotrophic
cultivation of mixotrophic microalgae strains for domestic wastewater
treatment and lipid production under dark condition. Bioresour. Technol. 149,
586589.