Bioresource Technology 174 (2014) 274280

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

Selection of microalgae for biodiesel production in a scalable outdoor

photobioreactor in north China
Ling Xia a, Shaoxian Song a, Qiaoning He b,c, Haijian Yang b,c, Chunxiang Hu b,
a

School of Resources and Environmental Engineering, Wuhan University of Technology, Wuhan 430070, China
Key Laboratory of Algal Biology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China
c
University of Chinese Academy of Sciences, Beijing 100049, China
b

h i g h l i g h t s
 Eight algal species were analyzed for growth, lipid accumulation and FA proles.
 Closed sterile culture and open culture were used for strain selection.
 Culture for strain characterization was scaling up.
3

 Average bio-oil productivity of 22.8 m ha

1

yr1.

 S. obtusus XJ-15 showed high capacity for biofuel production in north China.

a r t i c l e

i n f o

Article history:
Received 13 August 2014
Received in revised form 27 September 2014
Accepted 1 October 2014
Available online 14 October 2014
Keywords:
Desmodesmus
Scenedesmus
Photobioreactor
Scale up
Biodiesel

a b s t r a c t
The aim of this study was to identify the most promising species as biodiesel feedstock for large-scale
cultivation in north China. Eight species of microalgae, selected on the basis of indoor screening, were
tested for lipid productivity and the suitability of their fatty acid proles for biodiesel production under
outdoor conditions. Among them, three species Desmodesmus sp. NMX451, Desmodesmus sp. T28-1 and
Scenedesmus obtusus XJ-15 were selected for further characterization due to their possessing higher lipid
productivities and favorable biodiesel properties. The best strain was S. obtusus XJ-15, with highest
biomass productivity of 20.2 g m2 d1 and highest lipid content of 31.7% in a culture of 140 L. S. obtusus
XJ-15 was further identied as the best candidate for liquid biofuel production, characterized by average
areal growth rate of 23.8 g m2 d1 and stable lipid content of above 31.0% under a scale of 1400 L over a
season.
2014 Elsevier Ltd. All rights reserved.

1. Introduction
Biodiesel have recently attracted extensive interests as it is carbon-neutral and environment-friendly. Among various sources for
biodiesel production, microalgae are considered as the most promising feedstock for the future of biofuel production because they
have high photosynthetic efciency, high growth rate, and can be
cultivated on non-arable land (Chisti, 2007). Moreover, the growth
of microalgae at the same time will contribute to Greenhouse Gas
savings (Wang et al., 2008).
In spite of many advances, producing microalgal oil for biodiesel is still too expensive. In fact, the rst step in an algal process is
to choose the right alga with relevant properties. However, the
robust algal growth and high lipid production are reversely related
Corresponding author. Tel./fax: +86 27 68780866.
E-mail address: cxhu@ihb.ac.cn (C. Hu).
http://dx.doi.org/10.1016/j.biortech.2014.10.008
0960-8524/ 2014 Elsevier Ltd. All rights reserved.

(Rawat et al., 2013). Certain strains of microalgae, such as Botryococcus braunii, have high lipid storage potential (75% by dry cell
weight (DCW)) but this is accompanied by low biomass productivity (Mata et al., 2010; Rawat et al., 2013). Due to this contradiction,
lipid productivity showed a combination of biomass productivity
and lipid production was considered as the most important selection parameter (Grifths and Harrison, 2009).
The appropriate oil quality, besides its yields, is also key desirable characteristic of algal-based biodiesel industry because it
inuences the efciency of biodiesel conversion and its quality
(Nascimento et al., 2013; Rawat et al., 2013; Talebi et al., 2013).
For example, Nannochloropsis, as one of the most promising sources
of oil feedstock for biodiesel production, has high biomass production capacity and relatively high lipid content (Rodol et al., 2009);
however, being rich in long chain polyunsaturated fatty acids
(PUFA) (Doan et al., 2011; Grifths et al., 2012), which is not desirable for biodiesel properties (e.g., ignition quality and oxidative

275

L. Xia et al. / Bioresource Technology 174 (2014) 274280

stability) (Doan et al., 2011). Thus, it should be analyzed

thoroughly.
In addition, to minimize costs, photoautotrophic microalgal biodiesel production must rely on freely available sunlight. Qualities
generally desirable for outdoor mass culture for microalgal lipid
production include resistance to contamination, tolerance to a
wide range of environmental conditions (above all temperature
and solar radiation changes), rapid CO2 uptake and tolerance to
shear force (Grifths et al., 2012). Various oil-rich microalgae, particularly those belonging to the genera Chlorella and Scenedesmus
have been considered as potential sources of renewable energy
(Ho et al., 2010; Song et al., 2013). Especially, Scenedesmus have
often been used for photosynthetic CO2 reduction combined with
biodiesel production (Ho et al., 2010; Toledo-Cervantes et al.,
2013). In addition, Desmodesmus, which are identied as thermotolerant genus, have recently drawn many attentions (Pan et al.,
2011). Thus, eight microalgae strains in this study belong to Scenedesmus or Desmodesmus were used to examine the capacities of oil
production under outdoor conditions.
Moreover, not all the oil-rich microalgae can be cultivated
under outdoor conditions in certain area. For example, oleaginous
microalgae Tetraselmis suecica CS-187 and Chlorella sp. were successfully realized a long-term outdoor cultivation in Victoria, Australia, but Dunaliella tertiolecta CS-175 failed to scale up after many
tries (Moheimani, 2012). In this study, an algal screening system
was rst attempted to expand culture and maintain a large scale
cultivation using the selected microalgae in north China.
The experiments were all conducted in a greenhouse in Beijing,
China using sunlight as energy source. The objective of this study
was to determine whether the eight green algal species had the
potential to accumulate lipid and select the most promising species for large scale cultivation. Biomass and lipid productivities,
lipid proles and the estimated biodiesel properties were selected
as critical factors for the evaluation. The selected robust species
were further examined via a cultivation of 140 L in the columns
to decide the best strain for biodiesel production. Moreover, this
study describes the culture of the selected strain in a larger scale
of 1400 L over a period of three months.

et al. (2013, 2014) in the green house in Beijing, China (40220 N,

116200 E). For strain selection, the cultures were grown in 5 L
Erlenmeyer ask (0.37 m height  0.22 m in diameter) with 3 L
medium. BG-11 was prepared from tap water which was ltered
through 1 lm polypropylene lters (FTW) or autoclaved distilled
water (ADW) and then added with nutrient solutions, respectively
for open culture and closed culture. The closed culture was sealed
with cotton plug and aerated continuously with sterile ltered air,
while open culture sealed with lter paper and aerated with air
without any treatment. The closed one kept sterile operation during the whole period of cultivation while the open one did not. To
ensure well cell mixing, a 5 cm magnetic stir bar (mixing at
150 rpm) was placed at the middle of the bioreactor chamber for
stirring. CO2 in air (0.035%, v/v) was supplied to each bioreactor
using an air compressor through the pipage during the daytime.
The following large scale trials using the selected microalgae were
carried out in 140 L bioreactor, which composed with two connected 70 L hanging column bags (1.80 m height  0.22 m in diameter). CO2 in air (0.035%, v/v) was supplied to each bioreactor
using an air compressor through the pipage during the daytime.
For all the trials, the initial cell concentration was 0.1 g L1, and
culture media was BG-11 with nitrogen source of only 0.2 g L1
urea because urea is signicantly less expensive and exhibited
more favorable effect on algal growth in outdoor cultures (Xia
et al., 2013, 2014). The average day time high and night time low
air temperatures during the tested period were 39 C and 23 C.
2.3. Analytical procedures
2.3.1. Biomass measurement
The biomass of 500 mL culture cells was harvested by centrifugation, then the wet cell mass was frozen overnight at 70 C and
freeze-dried at 54 C under a vacuum (Alpha 1-2 LD plus, Christ).
The biomass concentration (BC, mg L1) of the tested microalgae
was determined by measuring optical density of 680 nm (OD680)
via an ultraviolet photospectrometer and using the following
equations:

BC D:abundans T12 320  OD680 R2 0:996

2. Methods

BC Desmodesmus sp: T28-1 322  OD680 R2 0:999

2.1. Organisms and culture

BC Desmodesmus sp: NMX451 230  OD680 R2 0:998

The eight microalgal species (Desmodesmus abundans T12, Desmodesmus sp. T28-1, Desmodesmus sp. NMX451, Desmodesmus
intermedius HB12-2, Desmodesmus obtusus XJ-36, Scenedesmus pectinatus var XJ-1, Scenedesmus obtusus XJ-15, Scenedesmus obtusus
XJ-19) used in this study, gifted by prof. Xu Xudong of Institute
of Hydrobiology in China, was selected after a laboratory screening.
Among these, D. abundans T12 and Desmodesmus sp. T28-1 were
isolated from one of the largest Chinese freshwater lake (Lake Taihu). Desmodesmus sp. NMX451 and S. obtusus XJ-19 were isolated
from Inner Mongolia of China. The rest of the species were isolated
from reservoirs or ponds in Hubei province in China. Stock cultures
for all the strains were grown in modied BG-11 medium containing 300 mg NaNO3, 30 mg K2HPO4, 36 mg CaCl22H2O, 6 mg ammonium citrate monohydrate, 6 mg ammonium ferric citrate, 1 mg
EDTA, 2.86 lg H3BO3, 1.81 lg MnCl24H2O, 0.222 lg ZnSO47H2O,
0.39 lg NaMoO45H2O, 0.079 lg CuSO45H2O, 0.050 lg CoCl26H2O
in 1 L sterile distilled water.

BC D:intermedius HB12-2 213  OD680 R2 0:998

BC D:obtusus XJ  36 201  OD680 R2 0:997

BC S:pectinatus v ar XJ-1 277  OD680 R2 0:998

BC S:obtusus XJ-15 229  OD680 R2 0:996

2.2. Experimental setup

All experiments were conducted in bioreactors (5 L ask bioreactor or 140 L airlift bag columns) as described previously by Xia

BC S:obtusus XJ-19 368  OD680 R2 0:999

8
1

The volumetric biomass productivity (VBP, mg L

culated according to the Eq. (9):

VBP B2  B1 =T

1

) was cal-

where B2 and B1 represents the dry weight biomass density at the

time T (days) and at the start of the experiment, respectively.
The areal biomass productivity (ABP, g m2 d1) was calculated
according to the Eq. (10). The calculation was based on VBP and the
oor area (required for columns and additional area required for
operational convenience such as empty space between reactors
as well as ground area to avoid shading effect), which is 0.26 m2
for each column.

276

ABP VBP  70=0:26  1000

L. Xia et al. / Bioresource Technology 174 (2014) 274280

10

2.3.2. Lipid analysis

The total lipid was gravimetrically quantied after extraction
using a Soxhlets extractor with chloroform/methanol (1/2, v/v)
as solvent and incubated at 90 C for 4 h. Lipid class separation
was performed by silica gel column chromatography. Solvent
sequences were as following: 6 volumes of chloroform to collect
the neutral lipid factions and 6 volumes of methanol for polar
lipids.
The lipid productivity (LP, mg L1 d1) was determined according to the Eq. (11):

LP BP  LC

11

where LC denotes the total lipid content (w/dry weight biomass, %)

at time T.
Biodiesel was determined as fatty acid methyl esters (FAME)
after acidic transesterication of lipids. In brief, 100 mg of lipid
sample was suspended in 1 M H2SO4methanol (2 mL) in a vial.
The vial was ushed with nitrogen to ensure an inert atmosphere
before sealing and heated at 100 C for 1 h in a water bathe. After
methylation, puried water (0.20 mL) and n-hexane (0.60 mL)
were added, the mixture centrifuged and the top hexane layer
contained FAME were collected. 1 lL FAME in hexane was
injected into gas chromatograph mass spectrometry (GCMS;
Thermo Scientic ITQ 700, USA) for analysis. All parameters of
the FAME were derived from the calibration curves generated
from the FAME standard mix (Supelco 37 component FAME mix,
SigmaAldrich).
Generally, the direct measurement of critical biodiesel quality
requires large amounts of oil and specialized devices, which are
not always available. Using the predictive models based on FA
composition makes the estimation of fuel property easy and
quick, which is of importance in biodiesel-based strain selection
(Nascimento et al., 2013; Talebi et al., 2013; Song et al., 2014).
The most important feedstock-related properties of biodiesel
were found to correlate with composition of the FA, providing
also linear best-t curves (Hoekman et al., 2012; Nascimento
et al., 2013; Talebi et al., 2013). The accuracy of these empirical
equations in estimating the quality of biodiesel has been previously shown (Ramos et al., 2009; Hoekman et al., 2012). Moreover, measured values and calculated values by these equations
in green algae C. protothecoides (Xu et al., 2006), Neochloris oleabundans (Gouveia et al., 2009) and S. obliquus (Abd El Baky
et al., 2012) were compared prior to this study, and results indicated that the values from equations of Song et al. (2014) were
very close to the measured values. Thus, the biodiesel property
in this study, the degree of unsaturation (DU), kinematic viscosity
(KS), specic gravity (SG), cloud point (CP), iodine value (IV),
cetane number (CN) and higher heating value (HHV) were determined by empirical equations from FA composition as described
by Song et al. (2014), while cold lter plugging point (CFPP) by
Nascimento et al. (2013).
2.3.3. Mass balance estimation
A simple mass balance from a culture scale of 1400 L is applied
according to Khoo et al. (2011) as follows:
? From the average total lipid content of 31.5% in algal biomass
and 51.7% triacylglycerol (TAG) content in total lipid, 1 kg of
dry microalgae biomass can produce a theoretical maximum
of 0.16 kg of TAG.
? From 1 kg TAG produced, assuming 90% is converted to biodiesel. Therefore the total amount of dry algal biomass required to
produce 1 kg biodiesel = 6.8 kg dry biomass.

3. Result and discussion

3.1. Strain selection
3.1.1. Growth and lipid accumulation properties of eight green algae
In any algal process, species selection is a key decision inuencing choice of location, reactor design, culture conditions, harvesting method and product range (Grifths et al., 2012). The
trials of strain selection were performed using autoclaved distilled water (ADW) and ltered tap water (FTW). The former
was conducted in the closed sterile culture and the later open
culture. The closed one was used to test the ability of algal cells
to tolerating environmental changes, especially in light intensity
and temperature. The open one was to further examine the resilience ability of cells to contamination. Eight freshwater microalgal strains (Desmodesmus and Scenedesmus) were evaluated for
their lipid production potential by evaluating their biomass productivity and lipid content under the above cultivation conditions (Table 1). After 15 days of batch cultivation, the cultures
reached the highest biomass productivity, and harvested for lipid
analysis.
Both average biomass productivity and lipid productivity of the
eight microalgal strains under ADW (BP, 200.3 mg L1 d1; LP,
45.4 mg L1 d1) were higher than that under FTW (BP,
163.7 mg L1 d1; LP, 38.1 mg L1 d1). In addition, the lipid contents in all the strains were also higher under ADW than that under
FTW, except for Desmodesmus sp. T28-1 and D. obtusus XJ-36. These
results indicate that water quality is an important factor affecting
the production capacity of microalgae, and should be considered in
the strain selection. The highest lipid content occurred in S. obtusus
XJ-15 under ADW; the value (31.31%) is higher than the literature
data in Scenedesmus indoors (Grifths and Harrison, 2009;
Nascimento et al., 2013). The lipid content was still increasing
after 15 days and may have reached higher values if cultivation
had been continued. Moreover, biomass and lipid productivities
of Desmodesmus sp. T28-1 and D. abundans T12 were much
higher under FTW. The results suggest that Desmodesmus sp.
T28-1 and D. abundans T12 showed excellent weatherability and
anti-contamination properties.
Lipid productivity has been suggested as the most appropriate
kinetic parameter for the comparison of the species for biodiesel
production (Grifths and Harrison, 2009). Though in the same
class, the 10 microalgal species showed different abilities to accumulate lipid, which indicated the fundamental importance of algal
species screening and evaluation for lipid production. In general,
the high lipid accumulation and robust biomass production were
mutually exclusive (Rawat et al., 2013). But under outdoor conditions, considering the weather factors, S. obtusus XJ-15 had the
highest biomass productivity and, at the same time the highest
lipid content. The result suggest that strain selection under outdoor conditions was very important.
The maximum lipid productivity was reached by S. obtusus
XJ-15 under ADW, and by Desmodesmus sp. T28-1 under AFW,
followed by D. abundans T12. Thus, S. obtusus XJ-15, Desmodesmus
sp. T28-1 and D. abundans T12 presented the highest lipid productivity and emerged from the screening as the best candidates for
algal oil production.

3.1.2. Fatty acid proling and estimated biodiesel properties of eight

green algae
GC proles for algal strains under FTW used in this study are
tabulated in Table 2. As seen, the C16-C18 FA, which is the most
common feedstocks suitable for biodiesel, possessed over 97% of
total FA in the eight green microalgae tested. The predominant
FAMEs found in all the algal oils were C16:0 and C18:1, with the

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L. Xia et al. / Bioresource Technology 174 (2014) 274280

Table 1
Biomass and lipid productivity of 8 microalgal strains cultivated in 5 L asks outdoors.
Strains

D. abundans T12
Desmodesmus sp. T28-1
Desmodesmus sp. NMX-451
D. intermedius HB12-2
D. obtusus XJ-36
S. obtusus XJ-15
S. obtusus XJ-19
S. pectinatus var XJ-1

Biomass concentration (g L1)

Lipid content (%)

Biomass productivity (mg L1 d1)

Lipid productivity (mg L1 d1)

ADW

FTW

ADW

FTW

ADW

FTW

ADW

FTW

2.6
2.7
2.4
2.6
2.6
3.6
2.4
2.8

3.0
3.2
2.0
1.9
1.6
3.3
3.0
1.8

25.1
24.6
25.5
24.6
23.1
31.3
22.7
23.2

23.2
26.6
23.2
23.3
24.1
24.1
18.5
22.6

171.5
180.0
160.1
169.7
170.3
238.4
160.0
183.5

199.0
210.9
134.0
125.9
104.8
222.7
193.3
119.2

43.3
44.3
40.8
41.8
39.3
74.6
36.4
42.5

46.2
56.1
31.1
29.3
25.5
53.7
35.8
26.9

ADW, autoclaved distilled water; FTW, ltered tap water.

Table 2
Fatty acid prole (%) of biodiesel from eight green algal oils in 5 L asks outdoors using tap water.
D.
abundans
T12

Desmodesmus
sp. T28-1

Desmodesmus sp.
NMX-451

D. intermedius
HB12-2

D. obtusus
XJ-36

S. obtusus
XJ-19

S. obtusus
XJ-15

S. pectinatus
var XJ-1

Saturated fatty acids (% of

total FAME)

C14:0
C16:0
C18:0
C20:0
Subtotal

0.94
3.30
11.01
1.32
16.57

0.24
46.06
6.18
1.20
53.68

0.18
56.71
8.20
0.74
65.83

0.49
39.33
6.01
0.61
46.44

0.68
43.98
6.09
ND
50.75

1.19
46.25
3.07
ND
50.51

0.56
39.82
6.11
ND
46.49

0.33
38.64
3.54
0.36
50.51

Monoenoic fatty acids (%

of total FAME)

C14:1
C16:1
C18:1
Subtotal

0.10
3.12
57.48
60.70

0.01
3.98
38.41
42.40

0.03
2.2
29.01
31.24

0.01
3.55
40.56
44.12

0.11
0.04
28.86
29.01

0.43
6.57
30.87
37.87

0.10
4.14
29.12
33.36

0.02
0.14
38.7
37.45

Polyenoic fatty acids (% of

total FAME)

C18:2
C18:3
C18:4
Subtotal

9.18
13.56
ND
22.74

2.46
1.37
0.09
3.92

1.66
1.19
0.08
2.93

5.66
3.49
0.30
9.45

9.81
9.48
0.93
20.22

5.41
5.84
0.37
11.62

9.74
9.52
0.87
20.13

9.86
7.85
0.57
6.20

C16C18 (% of total FAME)

C16
C18

97.64

98.46

98.97

98.59

98.28

98.02

98.47

98.72

ND: not detected.

individual amounts varying signicantly. For example, Desmodesmus sp. NMX451 possessed the highest content of C16:0 (56.71%
DCW), while D. abundans T12 contained rather low content of
3.30%. Relatively, D. abundans T12 possessed the highest content
of C18:1 (57.48% DCW), while D. obtusus XJ-36 contained rather
low content of 28.86%. The peaks for Palmitic acid (C16:0) and oleic
acid (C18:1) in Desmodesmus and Scenedesmus were consistent
with the observations by Nascimento et al. (2013) and Song et al.
(2014). In addition, highly unsaturated FA, C20C24 with 36 double bonds, appear to be more commonly occurring in the FA of algal
oils but barely detected in the eight microalgae (Doan et al., 2011;).
The consensus view is that the most favorable biodiesel would
have rather low levels of polyunsaturated (PUFA) and low levels
of saturated FA (SFA) to decrease oxidative stability and low temperature property (Knothe, 2009). However, the rather high lipid
content of SFA (C16:0) in the tested strains would resulted in cold
ow problems. Nevertheless, the feedstock rich in SFA have higher
CN than fuels produced from less saturated feedstock (Knothe,
2009; Hoekman et al., 2012). Moreover, SFA and PUFA can easily
be converted to monounsaturated FA (MUFA) when changing the
culture conditions in green algae (Grifths et al., 2012). In addition,
oxidative stability is determined not only by FAME composition,
but also by the age of the biodiesel and the conditions under which
it has been stored (Hoekman et al., 2012). In addition, the carbon
carbon double bond orientation is also important for the trans conguration is more stable than cis (Hoekman et al., 2012). Although
some strains in this study, such as S. obtusus XJ-15, had a high content of PUFA (20%), its oxidative stability required further
investigation.

Table 3 presents the most important properties of biodiesel predicted based on FA proles for the 8 microalgae strains studied as
well as for oil-rich microalgae Nannochloropsis sp. and traditional
crops rape and soybean. All the microalgae tested satised the criterion set by the Chinese National Standards (Table 3). Moreover,
according to the two most common quality standards for biodiesel,
ASTM D6751 in the US and EN 14214 in Europe, the values of kinematic viscosity, specic gravity, cetane number and iodine number
of the eight candidates satised the specications, except for D.
abundans T12. The oxidative instability factor IV had a high value
beyond the standard in D. abundans T12 as well as in Nannochloropsis sp. and soybean (Table 3). However, biodiesel property of D.
abundans T12 can be improved when subjected to high salt or high
alkaline (Xia et al., 2014). The CN is a prime indicator of biodiesel
quality, and related to the ignition delay time and combustion
quality. The higher the CN is, the better is in its ignition properties.
CN in all the tested species showed a higher value than that in Nannochloropsis sp. and soybean (Table 3), which could help ensure
better cold start properties and minimize the formation of white
smoke. Similar ndings also have been observed in other studies
(Grifths et al., 2012; Talebi et al., 2013). There are no denite
specications of cloud point (CP) and cold lter plugging point
(CFPP), due to the different climate condition in the United States
and Europe (Knothe, 2011). Nonetheless, all of the species examined met the winter specication, even for a relatively warm country such as Spain (10 C; Ramos et al., 2009) or South Africa
(3 C; Stansell et al., 2011).
It is noticeable that Desmodesmus sp. NMX51 showed the best
biodiesel property with the highest CN value and the lowest IV


Report

3.2. Growth of three selected microalgae in a 140 L airlift column

bioreactor

39.73
16.09

40.03
16.00

40.02
14.95

41.19
15.32

5

Report

The environmental factors including light intensity, temperature and pH value uctuated during the cultivation. Of the above
parameters, light intensity was affected by the weather most.
When the weather was clear, the light intensity on the surface of
the column was as high as 500 lmol photons m2 s1. And once
in rainy days, the illuminations were close to zero. The haze in
north China was another important factor affecting the illumination. The temperature changed with the illumination but the
degree of variation was smaller than the light intensity. Temperature of the medium was controlled at a value under 32 C with cold
system in the green house at daytime, and the temperature at
night was at 2028 C. The pH value was regulated by the aeration
of CO2. The CO2 concentration in the atmosphere aerated was
depending on the weather. When it is sunny, the concentration
of CO2 was as high as 5%, and when cloudy or rainy, under 1%.
The pH value of the medium during cultivation was controlled at
810, which allowed a robust growth of microalgal cells and also
could help resist contamination such as zooplankton and microorganisms because of the alkaline environment (Bartley et al., 2014).
The growth and lipid accumulation properties of the selected
elite microalgae Desmodesmus sp. NMX451, Desmodesmus sp.
T28-1 and S. obtusus XJ-15 in the 140 L bioreactors were summarized in Table 4. The trials were conducted for 13 days, but the
effective illumination time only lasted for 9 days. And during the
foggy and wet days with little illumination, the cells barely grew.
Although all of the three species studied survived during the
weather changes and recorded a high yield, the biomass yield
decreased by about 25% (Table 4). Therefore, the yields from certain strain cultivated in certain area are very important and necessary, which provided data support to the true potential for the algal
biofuel production.
The actual areal biomass productivity obtained in both Desmodesmus sp. T28-1 and S. obtusus XJ-15 was above 20 g m2 d1.
Moreover, the lipid content in S. obtusus XJ-15 without lipid induction phase was as high as 31.0% of DCW, which met the commercial production of biodiesel production (Chisti, 2007). Therefore,
S. obtusus XJ-15 showed the best biodiesel potential and selected
for larger scale investigation.

39.71
14.09
39.21
13.43

39.96
15.97

128
125.08
101.32
101.68
88.86
88.43
67.13

98.63

0.88
8.29
58.40
0.87
8.31
60.31

0.88
8.28
57.49

0.88
8.29
58.36

0.88
8.28
57.21

0.88
8.28
57.25

0.86
0.18
52.80

49

0.850.90

Min 47

Min 49

1.96.0

3.5
5.0

Min
51
Max
120

Report

1.96.0
1.44
4.20
1.51
4.25
0.84
4.67
0.85
4.67
0.68
4.78
0.81
4.70
0.67
4.78
0.38
4.96

S. obtusus
XJ-15
S. obtusus
XJ-19
D. obtusus
XJ-36
D. intermedius
HB12-2
Desmodesmus sp.
NMX-451

value. Therefore, the traits of contamination toleration, high lipid

productivity and good biodiesel property potential make S. obtusus
XJ-15, Desmodesmus sp. T28-1 and Desmodesmus sp. NMX51 suitable candidates for bioenergy production and selected for further
analysis.

39.45
12.19
40.64
11.96

Based on Doan et al. (2011).

Ramos et al. (2009).
a

77.06

0.88
8.30
59.42
0.88
8.25
54.89

127.58

Iodine number (g I2
100 g1)
HHV (MJ/kg)
CFPP (C)

0.52
4.88
1.20
4.45

Avg. unsaturation
Kinematic viscosity
40 C (mm2 s1)
Specic gravity (kg L1)
Cloud point (C)
Cetane number

Desmodesmus
sp. T28-1

3.3. Cultivating S. obtusus XJ-15 under a scale of 1400 L

D.
abundans
T12

Table 3
PredictedEstimated properties of biodiesel based on fatty acid proles from eight green algal oils in 5 L asks outdoors using tap water.

S. pectinatus
var XJ-1

Nannochloropsis
sp.a

Soybeanb

ASTMD6751

GB/T
208282007

L. Xia et al. / Bioresource Technology 174 (2014) 274280

EN
14214

278

To further evaluate the potential of the selected elite microalgae

for biofuel production, S. obtusus XJ-15 was inoculated into a row
of connected columns in an area of 5.4 m2 with a scale of
1400 L. Table 5 presents the biomass and lipid productivities after
three runs of batch cultivation using untreated tap water during
June to September, 2013. In the rst and the third run, algae
recorded a volumetric biomass productivity of about 100 mg L1 d1,
corresponding to an areal biomass productivity of over
26 g m2 d1. However, the second run recorded a lower productivity of 68.6 mg L1 d1. This is largely due to the relatively low
light intensity in haze days during late July and early August
(Table 5). It was evident from the results that weather factors
especially light intensity directly inuences the outdoor yield of
microalgae (Moheimani, 2012; Hindersin et al., 2014). Therefore,
compensative articial light source is necessary and important
when the solar light intensity decreased to a value that could not

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L. Xia et al. / Bioresource Technology 174 (2014) 274280

Table 4
Biomass and lipid productivities of S. obtusus XJ-15, Desmodesmus sp. NMX451 and Desmodesmus sp. T28-1 cultivated in a 140 L photobioreactor.
Biomass concentration (g L1)

Desmodesmus sp. T281

Desmodesmus sp. NMX451
S. obtusus XJ-15
a
b

0.8 0.1
0.9 0.0
0.9 0.0

Volumetric biomass
productivity (mg L1 d1)

Areal biomass
productivity (g m2 d1)

Actuala

Theoreticalb

Actuala

Theoreticalb

75.4 7.6
68.7 3.2
75.6 3.6

97.0 9.8
103.0 10.4
98.1 9.4

20.1 2.0
18.3 1.1
20.2 1.0

25.9 2.6
27.5 2.0
26.1 2.5

Lipid content (%)

Lipid productivity (mg L1 d1)

21.7 0.7
27.06 0.8
31.7 1.5

21.0 0.2
27.87 1.2
31.0 2.8

Including foggy, cloudy and rainy days.

Clear days with effective illumination.

Table 5
Biomass and lipid productivities of S. obtusus XJ-15 in a scale of 1400 L.
Run

1
2
3

Biomass concentration
(g L1)

Lipid content
(%)

Biomass productivity
Volumetric
(mg L1 d1)

Areal
(g m2 d1)

1.1 0.3
0.8 0.1
0.9 0.0

31.7 1.5
31.4 0.8
31.4 0.8

98.1 9.4
68.6 11.7
100.8 4.9

26.1 2.5
18.3 3.1
26.9 1.3

support microalgal growth. However, the average biomass productivity of the three runs reached 23.8 g m2 d1, which is higher
than most of the oil-rich microalgae recorded in the literatures
(Chen et al., 2011; Nurra et al., 2014).
Moreover, the lipid content in the three runs maintained stably
at above 31%, with an average content of 31.5%. This lipid content
was much higher than most of the records in the outdoor cultures,
indicating a lipid content of 15.6% for Botryococcus braunii (Bazaes
et al., 2012), 1932% for Chlorella sp (Moheimani, 2012, 2013),
34.7% for N. oculata and N. salina (Quinn et al., 2012), less than
30% for S. acutus (Doria et al., 2011), and 2430% for Tetraselmis
suecica (Moheimani, 2013). In addition, the lipid productivity
averaged at 28.1 mg L1 d1, peaked at 31.7 mg L1 d1. The lipid
production achievable by S. obtusus XJ-15 is equivalent to
22.8 m3 ha1 yr1 for a 9-month production period. And this value
was two times higher against 10.7 m3 ha1 yr1 achieved by
Nannochloropsis (Quinn et al., 2012). Regarding the biomass productivity and the biomass balance estimation, it can be predicted
that the strain S. obtusus XJ-15 has the capability to produce
10,600 L ha1 yr1 of biodiesel. As for the rest cold winter days of
the year in north China, further work on exploring and culturing
other candidates which can grow and accumulate useful components in this period is needed to reduce the total cost for biodiesel
production. Thus, S. obtusus XJ-15 showed an outstanding capacity
for oil production, and it is suitable for microalgae cultured in
north China.
4. Conclusions
Eight microalgae were investigated for their potential for biodiesel production under outdoor conditions. In addition to comparing the two key characteristics (lipid productivity and fatty acid
prole) for promising microalgae species, this work highlighted
the importance of tolerances to water quality and environment
factors in screening microalgal strains. Through examination, S.
obtusus XJ-15 was nally selected and cultivated in a scale of
1400 L through three turns. Both high lipid content (31%) and high
lipid productivity (22.8 m3 ha1 yr1) were achieved. This study
assesses the importance and helps to ll the gap of the microalgae
cultivated in north China.
Acknowledgements
This work was supported by National 863 program
(2013AA065804) and Program of Sinopec, international partner

Lipid productivity
(mg L1 d1)

Light intensity (lmol photons

m2 s1)

Temperature
(C)

31.0 2.8
21.6 3.6
31.7 1.7

665.5
372.0
438.1

27.3
26.9
28.4

program of innovation team (Chinese Academy of Sciences), Platform construction of oleaginous microalgae (Institute of Hydrobiology, CAS of China).

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