Energy Conversion and Management 138 (2017) 556564

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Energy Conversion and Management

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Biodiesel production from waste cooking oil in a magnetically fluidized

bed reactor using whole-cell biocatalysts
Guanyi Chen a,b,c,1,, Jing Liu a,1, Jingang Yao a,d, Yun Qi a,, Beibei Yan a
a
School of Environmental Science and Engineering/State Key Lab of Engines, Tianjin University, Tianjin 300072, China
b
School of Science, Tibet University, Lhasa 850012, China
c
Tianjin Engineering Center of Biomass-derived Gas/Oil Technology, Tianjin 300072, China
d
Institute of Chemical Engineering, Vienna University of Technology, Vienna 1060, Austria

a r t i c l e i n f o a b s t r a c t

Article history: Biodiesel production from catalytic transesterification of waste cooking oil (WCO) was investigated in a
Received 14 December 2016 magnetically fluidized bed reactor (MFBR) over Pseudomonas mendocina cells immobilized in magnetic
Received in revised form 11 February 2017 microspheres. The effects of methanol to oil molar ratio (MOMR), magnetic field intensity, biocatalysts
Accepted 13 February 2017
concentration and reactant flow rate on biodiesel production were investigated. Optimization of the
selected parameters was carried out for maximum biodiesel production using response surface method-
ology with support of Design-Expert software. The parameters optimized with response surface method-
Keywords:
ology were MOMR of 3.74:1, magnetic field intensity of 136.63 Oe, biocatalysts concentration of 10.21 wt.
Biodiesel
Waste cooking oil
% and reactant flow rate of 16.97 mL/min. An experimental biodiesel yield of 91.8% was obtained at 35 C
Magnetically fluidized bed reactor after 48 h with these optimized parameters. Moreover, the magnetic whole-cell biocatalysts (MWCBs)
Whole-cell biocatalyst exhibited good reusability in MFBR that 87.5% biodiesel yield could still be achieved after 10 cycles.
Immobilization The results suggested that MWCBs catalyzed transesterification in the MFBR system would have broad
application prospects in biodiesel production.
2017 Elsevier Ltd. All rights reserved.

1. Introduction discharged to the environment. Therefore, conversion of WCO into

Biodiesel is considered as a biodegradable and renewable bio-
fuel that can be used in diesel engines with very less pollution
on the environment [1,2]. It has been reported that biodiesel can
provide a value in reducing greenhouse emissions and acid-rain
pollution [3]. Currently, biodiesel is primarily made from vegetable
oils or animal fats. Nevertheless, the production costs of biodiesel
from these costly edible vegetable oils are approximate 1.5 times
higher than that for petroleum-based diesel, preventing its large-
scale application [4]. To achieve sustainable low-cost biodiesel
production, using WCO as feedstock is deemed to be an ideal
option. According to literature, there was a decrease up to 70%
for production costs of biodiesel when using WCO [5]. The annual
production of WCO in China has been estimated up to 5 million
tons [6]. Inappropriate disposal of the WCO can result in significant
health, environmental or economic issues [79]. Even though WCO
can be applied to the soap production, large portion of it is
Corresponding authors at: School of Environmental Science and Engineering/
State Key Lab of Engines, Tianjin University, Tianjin 300072, China (G. Chen).
E-mail addresses: chen@tju.edu.cn (G. Chen), qiyun@tju.edu.cn (Y. Qi).
1
Both authors contributed equally to this work.
biodiesel as petrodiesel substitute has great practical significance
for increasing economic benefit, while minimizing the health and
environmental impacts.
Nowadays, biodiesel is primarily derived from oils using
chemical-catalyzed process. However, chemical methods suffer
from soap formation, side streams, glycerol recovery, wastewater
treatment and catalyst removal [1012]. Enzymatic transesterifica-
tion offers good solutions to these existing problems, but its com-
plex purification procedures and unreliability of lipase have
hampered the adoption of this technology [13]. In light of present
problems, researchers turn their focus towards the potential use of
microorganisms that could serve as whole-cell biocatalysts [14]. As
reported, a biodiesel yield of 82% was obtained from transesterifi-
cation of WCO by using Thermomyces lanuginosus lipase-displaying
Pichia pastoris as whole-cell biocatalysts after 84 h [15]. The
Bacillus subtilis cells were used to catalyze the transformation of
WCO and could give a biodiesel yield of 90% after 72 h [16]. The
recombinant Escherichia coli cells exhibited the reusability that
55% of its activity could still be retained even after 5 cycles [17].
Nevertheless, all these whole cell-catalyzed bioprocesses are still
on a laboratory scale. This reaction process also confronts with

http://dx.doi.org/10.1016/j.enconman.2017.02.036
0196-8904/ 2017 Elsevier Ltd. All rights reserved.
 G. Chen et al. / Energy Conversion and Management 138 (2017) 556564
many problems, such as separation, uneven distribution of biocat-
alyst, low mass transport and so on.
Developing effective reactors is of great importance, which can
help to improve the feasibility and reliability of biodiesel produc-
tion in large-scale transesterification catalyzed by whole-cell bio-
catalysts. Currently, MFBR has become attractive as an efficient
system coupled with MWCBs for biodiesel production due to its
ability to provide the external magnetic field, which keeps and sta-
bilizes the MWCBs in the MFBR. Besides, the MFBR provides many
advantages, such as low pressure drop through the reactor, higher
mass-transfer efficiencies, countercurrent contact and elimination
of solid mixing [1820]. MFBR makes it possible to process large-
scale MWCBs-catalyzed transesterification for biodiesel produc-
tion. Sodium alginate is becoming increasingly attractive as an
immobilized material with several advantages such as biocompat-
ibility, physiological inertness, high mechanical stability and non-
toxicity [19,21,22]. Introducing Fe3O4 in sodium alginate
microspheres is conducive to the formation of microspheres with
superparamagnetism. In this study, MWCBs were constructed by
immobilizing Pseudomonas mendocina cells into Fe3O4-sodium
alginate microspheres, characterized and investigated for produc-
ing biodiesel from WCO by using a MFBR. The effects of MOMR,
magnetic field intensity, biocatalysts concentration and reactant
flow rate on biodiesel production were investigated. Response sur-
face methodology (RSM) was applied in this work to optimize the
reaction conditions for maximum biodiesel production.
2. Materials and methods
2.1. Materials
P. mendocina cells were provided by China General Microbiolog-
ical Culture Collection Center. Agar powder, peptone and yeast
extract were purchased from Biosinc Technology Co., Ltd. (Tianjin,
China). The standard fatty acid methyl esters (FAMEs) were
obtained from Anpel laboratory technology Inc. (Shanghai, China).
FeSO4
7H2O, NaHCO3, CaCl2, FeCl3
6H2O, methanol, ethanol, poly-
ethylene glycol, glutaraldehyde and sodium alginate were obtained
from Yuxiang Technology Co., Ltd. (Tianjin, China). Oleic acid and
olive oil were obtained from Cofco Group Co., Ltd. (Tianjin, China).
2.2. Pretreatment of WCO
The WCO was collected from a local fast food restaurant in Tian-
jin, China. The WCO sample was first filtered to remove crude
impurities by using filter paper. Followed, 1% of 85% H3PO4 was
added in the filtrate, stirring at 80 C for 15 min; and another
30 min after adding equiponderant water. The mixture obtained
was then introduced into a separatory funnel so that impurities
(excess water and colloid) would settle down and then be removed
from WCO. Finally, the WCO was treated with activated clay, which
plays an important role in the decoloration of WCO. The fatty acid
analysis of WCO is summarized in Table 1.
2.3. Preparation of oleic acid-coated Fe3O4
FeCl3
6H2O (6.97 g) and FeSO4
7H2O (3.58 g) were dissolved in
150 mL polyethylene glycol solution (1.5 wt.%). Followed, 2 M
NaHCO3
solution was added, stirring at 80 C for 30 min. 5 mL oleic
acid was subsequently added into the solution, stirring vigorously
for 100 min. Finally, the magnetic precipitates were separated by
the magnet, washed 4 times with ethanol and water and dried in
a vacuum freeze dryer.
557
Table 1
The composition of fatty acid in WCO.
Fatty acid Content (wt.%) Fatty acid Content (wt.%)
composition composition
C14:0 1.36 C18:2 46.28
C16:0 0.88 C18:3 2.19
C18:0 7.32 C20:0 0.73
C18:1 39.60 Other components 1.64
2.4. Preparation of MWCBs
P. mendocina cells were incubated in Luria-Bertani liquid med-
ium at 35 C and 180 rpm for 1 day. Suspended P. mendocina cells
(>107 cells/mL) and sodium alginate solution (2%, w/v) containing
3.5% (w/v) oleic acid-coated Fe3O4 were mixed (1:8, v/v) thor-
oughly with the help of ultrasonic wave. The mixture was then
dripped into sterilized CaCl2 solution. Subsequently, 25 wt.% glu-
taraldehyde was added, stirring for 90 min. Finally, MWCBs pre-
pared were collected by magnetic separation and washed to
neutral by H2O. Apart from the need of introducing oleic acid-
coated Fe3O4, all other procedures for preparing Fe3O4-
uncontained whole cell biocatalysts were in accordance with those
for MWCBs preparation. Finally, Fe3O4-uncontained whole cell bio-
catalysts prepared were separated by filtration. The microscopic
counting method was adopted to calculate the numbers of cells
immobilized in microspheres [19,23].
2.5. Transesterification in MFBR system
Fig. 1 shows the experimental set-up of MFBR. A jacketed plex-
iglass column covered by coaxial coils was the core part of MFBR,
which could provide a homogeneous and axial magnetic field sur-
rounding the whole column. The inner diameter and length of the
column were 100 mm and 950 mm, respectively. The flow rate of
air pump was 0.25 cm/s. The intensity of magnetic field could be
adjusted by controlling the direct current passing through the
coils. For all experiments, methanol and WCO were used as feed-
stocks and were introduced into MFBR by pumps. Besides, the
reaction liquid in MFBR was circulated continuously by a peri-
staltic pump. The reaction temperature was adjusted by the
constant-temperature water circulator. The bioreactor column
was filled with P. mendocina cells immobilized magnetic micro-
spheres. The total force of magnetic microspheres received in
MFBR was calculated by Eq. (1):
X
F Fm Fg  Fd  Ff 1
where Fm, Fg, Fd and Ff were magnetic force, gravity, drag force and
flotage, respectively. As observed, the motion of MWCBs could be
controlled by changing the intensity of magnetic field.
The effects of MOMR (1:1, 2:1, 3:1, 4:1, 5:1 and 6:1), magnetic
field intensity (60220 Oe), biocatalysts concentration (117 wt.%)
and reactant flow rate (540 mL/min) on biodiesel production
were investigated. The initial conditions were MOMR of 4:1, mag-
netic field intensity of 120 Oe, biocatalysts concentration of 12 wt.
% and reactant flow rate of 20 mL/min for 35 C, 48 h. In the single-
factor experiments, only one parameter was changed at a time
keeping the others constant. Furthermore, RSM coupled with
Box-Behnken design (BBD) was adopted to optimize the reaction
parameters for maximum biodiesel yield [24].
2.6. Analytical methods
2.6.1. Biocatalyst characterization
Micrographs were observed by a scanning electron microscopy
(SEM, SU-8000) and a transmission electron microscopy (TEM,
558 G. Chen et al. / Energy Conversion and Management 138 (2017) 556564
Fig. 1. The MFBR system (1. methanol tank; 2. oil tank; 3. peristaltic pump; 4. mixing tank; 5. preheating system; 6. power supply; 7. plexiglass column filled with MWCBs; 8.
air pump; 9. circulating water; 10. product-separating tank).
JEM-2100F). X-ray diffraction (XRD) patterns were obtained on a
D/MAX-2500 diffractometer. The magnetic characteristics were
analyzed by a vibrating sample magnetometer (VSM, LDJ-9600).
2.6.2. Lipase activity
The lipase activity of MWCBs was quantified by a method
reported in our previous work [25]. For MWCBs, one-unit lipase
activity was defined as the amount of lipase releasing 1.0 lmol
fatty acids per minute under the assay condition.
2.6.3. Analysis of FAMEs
The FAMEs content in the reaction mixture was quantified by an
Agilent 7890A gas chromatography. More details of the analysis
method can be found in the literature [26]. The biodiesel yield
obtained in this work was defined by the Eq. (2):
weight of biodiesel produced
Yield %  100%
theoretical maximum weight of biodiesel
3. Results and discussion
3.1. Biocatalyst characterization
SEM image of MWCBs is displayed in Fig. 2(a). The MWCBs were
spherical microspheres with diameters around 35 mm. A rough
and porous surface structure can be observed for MWCBs, indicat-
ing a successful immobilization of P. mendocina cells on the surface
of MWCBs. These results show that the cells were well immobi-
lized on the surface of magnetic microspheres and that some cells
penetrated pores of magnetic microspheres. These phenomena are
mainly attributed to sodium alginates cross-linking properties and
its flocculating function. TEM image of magnetic nanoparticles
(oleic acid-coated Fe3O4) is displayed in Fig. 2(b). As observed,
magnetic nanoparticles have diameters ranged from 7 to 15 nm.
The XRD pattern of MWCBs in Fig. 3(a) shows all characteristic
peaks of standard Fe3O4 (JCPDS card nos. 85-1436) and confirms
the presence of magnetic Fe3O4 nanoparticles in MWCBs [27].
The crystallite size of Fe3O4 calculated by Scherrer equation
(dXRD = kk/bcosh) is about 10.6 nm, which is in accordance with
results obtained from TEM. These results give straightforward evi-
dences to the superparamagnetic property of nanoparticles [28].
Fig. 3(b) presents magnetization curves of MWCBs at 300 K. As
observed, MWCBs exhibits an S-shape magnetization curve. The
saturation magnetization, coercivity and remnant magnetization
intensity of MWCBs were 39.1 emu/g, 0.0096 emu/g and
0.0047 emu/g, respectively. The extremely low values of coercivity
and magnetic remanence indicate a superparamagnetic property of
MWCBs [27]. These results clearly support the analyses from XRD.
As seen in Fig. 3(c), the lipase activity of MWCBs increases obvi-
ously as incubation time evolves. The lipase activity of MWCBs
reached 2960 U/g after 10 h. This provides evidence that the pro-
duced lipase was secreted outside of the P. mendocina cells. As
2
reported by other researchers, lipase activities of 309.5 U/g from
immobilized Aspergillus niger lipase [29], 1154 U/g from immobi-
lized Burkholderia sp. lipase [30] and 1581.5 U/g from immobilized
Schizophyllum commune lipase [31] were obtained, respectively,
which are all lower than the value obtained in this work.
3.2. Effect of MOMR
The effect of MOMR on MWCBs-catalyzed transesterification
was investigated by varying the MOMR (1:1, 2:1, 3:1, 4:1, 5:1
and 6:1). Fig. 4 illustrates the biodiesel yield as a function of time
under the initial conditions. For a given MOMR, the biodiesel yield
increased first and then reached a steady state in the range studied.
At a given reaction time (72 h), the biodiesel yield gives a trend of
first increase and then decrease by increasing the MOMR. When
MOMR was 4:1, the biodiesel yield reached a maximum value of
94.0% after 72 h. The decrease in biodiesel yield beyond the MOMR
of 4 can be explained by the fact that excessive methanol accumu-
 G. Chen et al. / Energy Conversion and Management 138 (2017) 556564 559

Fig. 2. (a) SEM micrographs of MWCBs and (b) TEM images of magnetic nanoparticles (Fe3O4).

Fig. 3. (a) XRD spectrum of MWCBs, (b) Magnetization curves of MWCBs at 300 K and (c) Time course of lipase activity of MWCBs.

lated around MWCBs can result in lipase inhibition by restructur-
ing the active centers, causing irreversible inactivation of MWCBs
[26,32]. As shown in Fig. 4, the biodiesel yield increased signifi-
cantly from 0 to 23% after 10 h at a MOMR of 1:1. This can be
explained by the high activity of MWCBs at lower methanol
concentration.
3.3. Effect of magnetic field intensity
Fig. 5 presents the effect of magnetic field intensity on transes-
terification in the magnetic field intensity range of 60220 Oe.
With increasing magnetic field intensity, the biodiesel yield shows
a trend of first increase and then decrease. When magnetic field
560 G. Chen et al. / Energy Conversion and Management 138 (2017) 556564

Fig. 4. Effect of MOMR on transesterification. Reaction conditions: magnetic field

Fig. 5. Effect of magnetic field intensity on transesterification. Reaction conditions:
intensity of 120 Oe, biocatalysts concentration of 12 wt.%, reactant flow rate of
MOMR of 4:1, biocatalysts concentration of 12 wt.%, 35 C and 48 h.
20 mL/min, 35 C.

intensity was 140 Oe, the biodiesel yield reached a maximum value
catalysts. These results clearly support that the magnetic field
of 90.8%. This trend is mainly attributed to the fact that the void
could help to stabilize the MWCBs in the MFBR.
volume in MFBR column can be adjusted by adding different mag-
netic field intensities [18,20]. At 60 Oe, magnetic field has little
3.5. Effect of reactant flow rate
effect on MWCBs, and magnetic microspheres flow freely in the
column. Thus, there is only a slight increase in biodiesel yield
The effect of reactant flow rate on MWCBs catalyzed transester-
between MWCBs and Fe3O4-uncontained whole cell biocatalysts
ification were studied in the reactant flow rate range of 540 mL/
catalyzed processes. As magnetic field intensity increased, MWCBs
min. As shown in Fig. 6(b), the biodiesel yield increases signifi-
were gradually magnetized and aligned along the magnetic field
cantly with rising the reactant flow rate from 5 to 20 mL/min,
direction. MWCBs with the best dispersion were obtained at 140
while the biodiesel yield decreases gradually with a further
Oe. The MWCBs can well spread in the whole MFBR column
increase in reactant flow rate. A high biodiesel yield of 91.0% was
because of no additional diffusion limitations. When the magnetic
obtained at a reactant flow rate of 20 mL/min. These variations
field intensity was over 140 Oe, agglomerates of MWCBs were
are mainly attributed to the fact that higher flow rate shortens con-
formed due to the presence of strong interparticle attractive forces.
tact time between the MWCBs and the reactants, and simultane-
Microsphere agglomeration can decrease the surface area of
ously, higher flow rate can lead to rupture of cell microspheres
MWCBs, resulting in a lower biodiesel yield. A fully compressed
due to shear, thus causing lower biodiesel yield. Similar phenom-
form of MWCBs was observed at 220 Oe, corresponding to a lower
ena have been reported [20,35].
biodiesel yield as compared to control group. These results verify
The biodiesel yield obtained from control experiment is signif-
that only appropriate magnetic field intensity can contribute to
icantly lower than that from MWCBs catalyzed process. These
good dispersibility of MWCBs in the MFBR column, thus producing
observations also support that the superparamagnetism property
more biodiesel.
of MWCBs played an important role in keeping its dispersibility
with the help of a magnetic field.
3.4. Effect of biocatalysts concentration
3.6. Optimization of reaction parameters using RSM
Fig. 6(a) presents the effect of biocatalysts concentration on bio-
diesel yield under given operating conditions (MOMR of 4:1, mag- Optimization of selected reaction parameters was carried out
netic field intensity of 140 Oe and reactant flow rate of 20 mL/min). for maximum biodiesel production using response surface
As shown in Fig. 6(a), the biodiesel yield increases with rising bio- methodology with support of Design-Expert 8.0.6 software (Stat-
catalysts concentration from 1 to 9 wt.%, while the biodiesel yield Ease Inc., USA). A three-level-four-factor BBD consisting of 27
almost keeps at a constant level by further increasing the MWCBs experimental runs was applied for fitting a second-order response
concentration. A maximum biodiesel yield of 90.6% was obtained surface. Table 2 shows the coded and actual values used in this
at a MWCBs concentration of 9 wt.% after 48 h. These are mainly work. The experimental project, the predicted values of response
attributed to the fact that excessive cell microspheres result in and experimental values were presented in Table 3.
higher viscosity and higher solid content in MFBR, leading to diffu- Using the designed experimental data (Table 3), BBD model was
sional limitations. Similar phenomena have also been reported by applied to develop a second-order polynomial model for describing
other researchers [33,34]. the relationships between the biodiesel yields and the variables.
Transesterification catalyzed by Fe3O4-uncontained whole cell The equation was illustrated as follows (Eq. (3)):
biocatalysts was performed under the same conditions and used
as a control. For a given condition, the control group gives a lower Y yield 91:60  10:84X 1  0:53X 2 1:10X 3  0:44X 4 0:35X 1 X 2
biodiesel yield than that of MWCBs. This could be attributed to the 1:08X 1 X 3 0:45X 1 X 4  0:23X 2 X 3  1:20X 2 X 4  0:98X 3 X 4
rupture of microspheres and cell leakage caused by irregular oscil-  25:84X 21  3:12X 22  1:53X 23  3:66X 24 3
lation and rotational motion of Fe3O4-uncontained whole cell bio-
 G. Chen et al. / Energy Conversion and Management 138 (2017) 556564 561

Fig. 6. Effects of biocatalysts concentration (a) and reactant flow rate (b) on transesterification. Reaction conditions: MOMR of 4:1, magnetic field intensity of 140 Oe, 35 C
and 48 h.

Table 2 Table 4
Variables and levels used for RSM design. Analysis of variance (ANOVA) for the regression model.

Variables Code Levels Source Sum of squares df Mean squares F-value P-value
1 0 1 Model 5457.11 14 389.79 200.43 <0.0001a
Residual 23.34 12 1.94
Methanol-to-oil molar ratio (mol/mol) X1 3:1 4:1 5:1
Lack of fit 23.34 10 2.33
Magnetic field intensity (Oe) X2 120 140 160
Pure error 0.000 2 0.000
Biocatalysts concentration (Wt.%) X3 7 9 11
Cor total 5480.45 26
Reactant flow rate (mL/min) X4 15 20 25
R2 = 0.9957; adjusted R2 = 0.9908; predicted R2 = 0.9755.
a
Significant at 1% level.
Table 3
Box-Behnken design for transesterification of WCO.
ability value (p-value) was less than 0.0001, suggesting a high sig-
Run Coded values of reaction Biodiesel yield (%)
nificance of the regression model and a reasonable fit of the
variables
predicted model with the experimental values [3638]. The coeffi-
X1 X2 X3 X4 Predicted Experimental cient of determination (R2) value was 0.9957. The adjusted R2 value
1 0 0 0 0 91.6 91.6 (0.9908) agreed well with the predicted R2 value (0.9755), indicat-
2 0 0 1 1 84.8 85.2 ing a close correlation between the predicted values and the
3 1 0 1 0 51.2 49.3
4 1 0 0 1 51.3 50.7
observed values. Therefore, the regression model is reliable to
5 1 0 0 1 72.1 71.5 reveal the mutual relationship of factors and to predict the response
6 0 1 1 0 90.9 90.2 values [39,40].
7 1 0 0 1 51.3 51.4 The interactions between the variables and biodiesel yield were
8 1 1 0 0 51.6 53.7
revealed very well by response surface plots (Fig. 7). Based on the
9 0 1 0 1 85.9 86.3
10 0 1 1 0 84.1 84.6 attained results and the equation, a maximum biodiesel produc-
11 0 0 1 1 88.9 89.5 tion of 92.0% is predicted by using the software. The final optimum
12 1 1 0 0 72.6 73.1 condition was as follows: MOMR of 3.74:1, magnetic field intensity
13 0 1 0 1 82.6 81.7 of 136.63 Oe, biocatalysts concentration of 10.21 wt.% and reactant
14 0 1 0 1 84.6 85.0
flow rate of 16.97 mL/min. The predicted value was validated
15 0 0 1 1 85.9 86.9
16 1 1 0 0 52.0 53.1 experimentally to verify the accuracy of the model. Under the
17 0 0 0 0 91.6 91.6 aforementioned optimal conditions, a biodiesel yield of 91.8%
18 0 0 1 1 86.1 87.3 was obtained, which were almost identical to the predicted value,
19 1 0 1 0 55.6 54.7
indicating reliability and good precision of the regression model.
20 1 0 0 1 73.8 72.6
21 0 1 1 0 87.6 87.2
22 1 0 1 0 75.1 76.5
23 1 0 1 0 75.1 75.4 3.7. Reusability of MWCBs in MFBR
24 1 1 0 0 74.3 73.9
25 0 1 1 0 85.2 83.6 Fig. 8 shows the time course of biodiesel yield under the oper-
26 0 1 0 1 86.1 85.2
ating conditions (MOMR of 3.74:1, magnetic field intensity of
27 0 0 0 0 91.6 91.6
136.63 Oe, biocatalysts concentration of 10.21 wt.% and reactant
flow rate of 16.97 mL/min, 35 C). As seen, a high biodiesel yield
where Yyield is the biodiesel yield; Xi (i = 1, 2, 3, 4) is the coded vari- of 91.8% is obtained by using MWCBs. MWCBs could be reused
ables. Here, the analysis of variance (ANOVA) for the regression for 5 times without significant decrease in biodiesel yield. More-
model (Table 4) was executed by this software. The calculated prob- over, a biodiesel yield of 87.5% could still be maintained even after
562 G. Chen et al. / Energy Conversion and Management 138 (2017) 556564

Fig. 7. Response surface plots of variables affecting biodiesel yield from WCO.

10 times of usage. In the MFBR, MWCBs can be magnetized with
the help of a magnetic field and then disperse along the magnetic
field lines because of dipole interaction [18]. Owing to their super-
paramagnetic property, the MWCBs were well dispersed and no
agglomeration was observed in the MFBR.
For the control group, the biodiesel yield decreased remarkably
after each cycle, and only 71.2% biodiesel yield retained after 10
times of usage. The decrease in biodiesel yield for the control group
can be attributed to rupture of microspheres and cell leakage
caused by irregular oscillation and rotational motion [18]. These
 G. Chen et al. / Energy Conversion and Management 138 (2017) 556564
Fig. 8. Reusability of catalysts for transesterification in MFBR. Reaction conditions:
MOMR of 3.74:1, magnetic field intensity of 136.63 Oe, biocatalysts concentration
of 10.21 wt.%, reactant flow rate of 16.97 mL/min and 35 C.
results also indicate that magnetic field can help to keep and stabi-
lize the MWCBs in the MFBR column. Given all that, using a MFBR
to perform MWCBs catalyzed transesterification has a great poten-
tial for large-scale biodiesel production.
4. Conclusion
A maximum biodiesel yield of 91.8% was obtained at 35 C after
48 h under the operating conditions (MOMR of 3.74:1, magnetic
field intensity of 136.63 Oe, biocatalysts concentration of
10.21 wt.% and reactant flow rate of 16.97 mL/min). The MWCBs
exhibited good reusability in MFBR that 87.5% biodiesel yield could
still be achieved after 10 cycles. The results suggested that MWCBs
catalyzed transesterification from WCO in the MFBR system would
have broad application prospects in biodiesel production.
Acknowledgments
This paper is financially supported by the Natural Science Foun-
dation of Tianjin, China (16JCYBJC20700), the National Energy
Administration of China (NY2013040302) and the China Scholar-
ship Council (CSC) (No. [2015]3022).
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