Anda di halaman 1dari 10

57 2007 Society of Chemical Industry and John Wiley & Sons, Ltd

Production of biodiesel:
possibilities and challenges
Sulaiman Al-Zuhair, UAE University, United Arab Emirates
Received 19 April 2007; revised version received 24 April 2007; accepted 10 May 2007
Published online 9 July 2007 in Wiley InterScience (www.interscience.wiley.com); DOI: 10.1002/bbb.2;
Biofuels, Bioprod. Bioref. 1:5766 (2007)
Abstract: Biodiesel, dened as monoalkyl fatty acid ester (preferentially methyl and ethyl esters), represents a
promising alternative fuel for use in compression-ignition (diesel) engines. Biodiesel fuel comes from renewable
sources as it is plant- not petroleum-derived and as such it is biodegradable and less toxic. In addition, relative to
conventional diesel, its combustion products have reduced levels of particulates, carbon oxides, sulphur oxides
and, under some conditions, nitrogen oxides. Enzymatic production of biodiesel has been proposed to overcome
the drawbacks of the conventional chemically catalyzed processes. The main obstacle facing full exploitation of the
enzyme, lipase, potential is its cost. Therefore, reuse of lipase is essential from the economic point of view, which
can be achieved by using the lipase in immobilized form. In addition, immobilized lipase displays improved stability
and activity. Common immobilization techniques include attachment to solid supports and entrapment within the
matrix of a polymer. This article presents a comparison between conventional processes and enzymatic processes
and different possible feedstocks for biodiesel production. In addition, possible ways to overcome the problems
facing the use of lipase are described. 2007 Society of Chemical Industry and John Wiley & Sons, Ltd
Keywords: biodiesel; lipase; alcohols; transesterication; immobilization; vegetable oils
Introduction
B
iodiesel is defned as the mono alkyl esters (methyl
and ethyl esters) of long chain fatty acids derived from
vegetable oils or animal fats, for use in
compression-ignition (diesel) engines. It is formed from
transesterifcation of vegetable oils with methanol (or
ethanol). Biodiesel has many merits as a renewable energy
resource that include being derived from a renewable,
domestic resource, thereby relieving reliance on petroleum
fuel, and it is biodegradable and non-toxic. Further, compared
to petroleum-based diesel, biodiesel has a more favourable
combustion emission profle, such as low emissions of carbon
monoxide, particulate matter and unburned hydrocarbons. In
addition, using biodiesel on a large scale will promote planta-
tions of crops used to produce its feedstock. Tis would result
in more carbon dioxide recycling by photosynthesis, thereby
minimizing the impact on the greenhouse efect.
1,2
Biodiesel
has a relatively high fash point (150
o
C), which makes it less
volatile and safer to transport or handle than petroleum
diesel.
3
It provides lubricating properties that result from
the free fatty acids present, which reduce engine wear and
Correspondence to: Sulaiman Al-Zuhair, Department of Chemical and Petroleum Engineering, UAE University,
17555 Al-Ain, United Arab Emirates. E-mail: s.alzuhair@uaeu.ac.ae
Review
58 2007 Society of Chemical Industry and John Wiley & Sons, Ltd | Biofuels, Bioprod. Bioref. 1:5766 (2007); DOI: 10.1002/bbb
S Al-Zuhair Review: Production of biodiesel: possibilities and challenges
extend engine life.
4
Furthermore, biodiesel has physical
properties and energetic content close to those of petroleum
diesel, which allows it to function ef ciently in conventional
diesel engines without any modifcation. Te author of this
review has investigated the use of biodiesels, from palm oil,
as an additive to conventional diesel fuel (CDF).
5
Te efects
of adding the biodiesel on fuel consumption rate and the
concentration of CO in ef uent gaseous emission were quan-
tifed. It was found that by using CDFbiodiesel emulsion a
considerable reduction in fuel consumption and CO concen-
tration was achieved when compared to pure CDF. At present,
the high cost of biodiesel is the major obstacle to its commer-
cialization. It has been reported, however, that the high cost of
biodiesel is mainly due to the cost of the virgin vegetable oil
used as feedstock.
3, 6

Conventional production of biodiesel
Te most common way to produce biodiesel is by transes-
terifcation, which refers to a catalyzed chemical reaction
involving vegetable oil and an alcohol to yield fatty acid
alkyl esters (i.e. biodiesel) and glycerol. Several aspects,
including the type of catalyst, alcohol/vegetable oil molar
ratio, temperature, water content and free fatty acid content
have an infuence on the course of transesterifcation. Trans-
esterifcation reactions are conventionally alkali-catalyzed
or acid-catalyzed.
Chemical catalysts production
Many studies of alkali-catalyzed transesterifcation have
been carried out and a reaction temperature near the boiling
point of the alcohol (e.g. 60
o
C for methanol) and a 6:1 molar
ratio of alcohol to oil were recommended.
79
Te kinetics of
the alkali-catalyzed system was studied
7, 8, 10
and the reac-
tion mechanism was formulated.
11
A commercial continuous
alkali-catalyzed transesterifcation process to produce methyl
esters on an industrial scale under high pressure (90 bar) and
at high temperature (240
o
C) was demonstrated by Kreutzer.
12

However, high energy consumption, a signifcant increase in
equipment cost and process safety issues related to the high
temperature, rendered this process prohibitive. Te most
important limitation of the alkali-catalyzed process is its
sensitivity to both water and free fatty acids contents in the
feedstock. Te presence of water may cause ester saponifca-
tion under alkaline conditions, and free fatty acids can react
with the alkali catalyst to produce soaps.
13, 14
Saponifcation
not only consumes the alkali catalyst, but also the resulting
soaps can cause the formation of emulsions, which creates
dif culties in downstream recovery and purifcation of
the biodiesel. Tus, dehydrated vegetable oil with less than
0.5 wt % free fatty acids, an anhydrous alkali catalyst and
anhydrous alcohol are necessary for commercially viable
alkali-catalyzed systems.
7, 15
Tis requirement is likely to be a
signifcant limitation to the use of a low-cost feedstocks.
16, 17
On the other hand, acid-catalyzed transesterifcation has
been largely ignored, mainly because of its relatively slower
reaction rate, despite its insensitivity to free fatty acids in the
feedstock. Freedman et al.
7
investigated the transesterifca-
tion of soybean oil with methanol using 1 wt % concentrated
sulfuric acid (based on oil). Tey found that at 65
o
C and a
molar ratio of 30:1 methanol to oil, it took 69 h to obtain
more than 90% oil conversion of methyl esters. Canakci and
Gerpen
18
studied the efects of the molar ratio of alcohol to
soybean oil, the reaction temperature, the amount of catalyst,
and the reaction time on ester conversion by acid-catalyzed
transesterifcation. Tey found that increased ester conver-
sions could be obtained at increased molar ratios of alcohol
to oil, increased reaction temperatures, increased concen-
trations of sulfuric acid, and longer reaction times. Te
kinetics of acid-catalyzed transesterifcation with butanol
was also investigated.
7
Studies of the acid-catalyzed system
have been very limited in number. No commercial biodiesel
plants have been reported to use the acid-catalyzed process.
Despite its relatively slow reaction rate, the acid catalyzed
process ofers benefts with respect to its independence from
free fatty acid content and the consequent absence of a
pre-treatment step. Tese advantages favour the use of the
acid-catalyzed process when using waste cooking oil as the
raw material, which usually has free fatty acids contents
greater than 2 wt %.
16, 17, 19
Non-catalytic production
A non-catalytic biodiesel production route with supercritical
methanol has been developed that allows a simple process and
high yield because of simultaneous transesterifcation of tria-
cylglycerols and methyl esterifcation of fatty acids.
20
It was
59
Review: Production of biodiesel: possibilities and challenges S Al-Zuhair
2007 Society of Chemical Industry and John Wiley & Sons, Ltd | Biofuels, Bioprod. Bioref. 1:5766 (2007); DOI: 10.1002/bbb
evident that at the subcritical state of alcohol, reaction rate is
very low and gradually increases as either pressure or temper-
ature rises. However, it was also observed that increasing the
reaction temperature, especially to supercritical conditions,
has a favourable infuence on the yield of ester conversion.
20

In addition, unlike the alkali-catalyzed method, the presence
of water afected positively the formation of methyl esters in a
supercritical methanol method. Comparison between chemi-
cally catalyzed processes and a non-catalytic supercritical
methanol method, for biodiesel production from vegetable
oils, is shown in Table 1.
Enzymatic production of biodiesel
Enzymes have been proposed to overcome the drawbacks
facing the conventional chemically catalyzed biodiesel
production methods, and have shown promising results.
Most importantly, glycerol can be easily recovered without
any complex process, free fatty acids contained in the oils
can be completely converted to methyl esters and subsequent
wastewater treatment is not required.
21

Lipase
Most lipases used as catalysts in organic synthesis are
of microbial and fungal origin, such as Candida rugosa,
Psuedomonas fuorescens, Rhizopus oryzae, Burkholderia
cepacia, Aspergillus niger, Termomyces lanuginosa and
Rhizomucor miehei. Microbial lipases are easy to obtain, by
fermentation processes and a few basic purifcation steps.
One important aspect of lipolytic enzymes is the unique
property of interfacial activation, identifed by Verger
et al.
22
Te activity of lipases is low in monomeric solutions
of lipid substrates but is strongly enhanced once organized
lipid structures and a confguration change of the enzyme
at the waterlipid interface are formed. Knowledge of the
molecular aspects of some catalytic steps is essentially based
on structural information obtained by X-ray crystallography,
site directed mutagenesis and classical biochemical methods.
X-ray studies of the 3D structure of lipase confrmed the
existence of two distinct domains in lipase: a large
N-terminal domain that contains the active site with a
catalytic triad formed by Ser, Asp and His, and a smaller
C-terminal domain.
23
Te active site is covered by a surface
loop completely buried and sitting directly on top of the
active site. At the interface, the lid domain is unfolded and
exposes the active sites. An experimental approach to deter-
mine the activation of the lipase at the interface, proposed
by Rooney and Weartherley,
24
was used by Al-Zuhair et al.
25

to determine that the activity of lipase at the interface was
15.7% higher than that in the bulk.
Pure lipases extracted from diferent sources have been
successfully used in the production of biodiesel,
17, 2640
as
shown in Table 2. C. antarctica B lipase, immobilized on
acrylic resin, commercially known as Novozym 435, was
by far the most commonly used enzyme for the production
of biodiesel. However, a comparative study on the type of
lipase powders from diferent sources
40
revealed that P. fuo-
rescens lipase showed the highest enzymatic activity. On the
other hand, Hama et al.
41
successfully used the whole-cell
R. oryzae as biocatalyst and investigated the efect of cell
membrane fatty acid composition on biodiesel-fuel
production.
Table 1. Comparisons between chemically catalytic processes and supercritical methanol method for biodiesel
production from vegetable oils by transesterification.
Alkali catalytic method Acid catalytic method Supercritical method
Reaction temperature (K) 303338 338 523573
Reaction pressure (MPa) 0.1 0.1 1025
Reaction time (min) 60360 4140 715
Methyl ester yield (wt %) 96 90 98
Removal of Methanol, catalyst Methanol, catalyst Methanol
Purication Glycerol, soaps Glycerol
Free fatty acids Saponied products Methyl esters, water Methyl esters, water
60 2007 Society of Chemical Industry and John Wiley & Sons, Ltd | Biofuels, Bioprod. Bioref. 1:5766 (2007); DOI: 10.1002/bbb
S Al-Zuhair Review: Production of biodiesel: possibilities and challenges
In addition to the type of lipase, other factors afecting the
efectiveness of lipase in the production of biodiesel from
triacylglycerol are the water content, temperature, number
of cycles (for immobilized lipase) and the type of alcohol and
its ratio to oil. Te efects of alcohol and number of cycles
are discussed later in this review.
Te efect of water content on the production of biodiesel
from soybean oil using lipases from R. oryzae,
31
C. rugosa
and P. fuorescens,
32
Novozym 435
42
and B. cepacia
34
have all
shown that enzyme activity was low in the absence of water,
which supports the fact that a minimum amount of water is
required to activate the enzyme. With increased addition of
water there was a considerable increase in ester production,
showing the enhancement in the activity of the enzyme.
However, the enzymatic activity gradually decreased at
higher water contents. Similar results were found in the
production of biodiesel from triolein using immobilized
P. fuorescens lipase.
40
Te author has shown that R. meihei
lipase maintains its activity up to 20% water,
39
which agrees
with other studies made on the same enzyme.
43
On the other
hand, methanolysis activity of Novozym 435 was inhibited
at a much lower water content of only 0.1% water.
42
Tis
efect of water on lipase activity is due to the fact that lipase
acts at the interface between aqueous and organic phases.
Activation of the enzyme involves unmasking and restruc-
turing the active site through conformational changes of the
lipase molecule, which requires the presence of an oilwater
interface.
23, 25
With increased addition of water, the amount
of water available for oil to form oilwater droplets increases,
thereby, increasing the available interfacial area. However,
since lipases usually catalyze hydrolysis in aqueous media,
excess water stimulates the competing hydrolysis reaction.
Table 2. Microbial lipases used for the production of biodiesel.
Lipase Oil Alcohol Optimum temperature (
o
C) Reference
Novozime 435 Soybean oil Methanol 17, 27, 32
Soybean oil Methyl acetate 26
Canola oil Methanol 38 28
Rice bran oil Methanol 29
Olive oil Methanol 40 30
Vegetable oil Methanol 33
Waste ABE Methanol, ethanol, 1-propanol, 34
1-butanol, iso-butanol,
iso-amylalcohol, and n-octanol
R. delemar Vegetable oil Methanol 33
R. miehei Vegetable oil Methanol 33
Palm oil Methanol 39
C. rugosa Waste ABE Methanol, ethanol, 1-propanol, 34
1-butanol, iso-butanol,
iso-amylalcohol, and n-octanol
Jatropha oil Ethanol 38
C. lipolytica Soybean oil Methanol 32
K. oxytoca Soybean oil Methanol 32
P. camembertii Soybean oil Methanol 32
P. uorescens Soybean oil Methanol 32
Triolein 1-propanol 60 40
Jatropha oil Ethanol 38
P. cepacia Soybean oil Methanol and ethanol 40 32, 34
Jatropha oil 38
61
Review: Production of biodiesel: possibilities and challenges S Al-Zuhair
2007 Society of Chemical Industry and John Wiley & Sons, Ltd | Biofuels, Bioprod. Bioref. 1:5766 (2007); DOI: 10.1002/bbb
Te optimum water content is a compromise between mini-
mizing hydrolysis and maximizing enzyme activity for the
transesterifcation reaction.
On the other hand, the efect of temperature on the
enzymatic production of biodiesel showed that as the
temperature is increased the reaction rate initially increases,
which is mainly due to an increase in rate constants with
temperature, and partly due to less mass transfer limitations.
However, the initial rate decreases sharply at the onset of
denaturation of the enzyme at higher temperatures. In addi-
tion to the deactivation of the enzyme, the presence of the
inactive enzyme at the interface blocks the active enzyme
from penetrating the interface, which further decreases
the reaction rate.
25
Tis trend was observed in all studies
that investigated the efect of temperature on the produc-
tion of biodiesel by lipase, however, the critical temperature
at which the enzyme starts to deactivate was diferent, as
shown in Table 2.
Feedstock
Vegetable oils
Several types of vegetable oils can be used for the prepara-
tion of biodiesel, as shown in Table 3. However, there are no
technical restrictions on the use of other types of vegetable
oils. Global vegetable oil production has increased from
56 million tons in 1990 to 88 million tons in 2000, and the
increase is still continuing.
11
Leading the gains in vegetable
oil production is palm oil, which increased from 17.1
million tons in 1997 to 19.3 million tons in 1999. In addi-
tion, palm oil has the highest yield of around 4000 kg per
hectare compared to that of other vegetable oils, the highest
of which, for coconut oil, is around 2250 kg per hectare.
Furthermore, greater palm oil yields, up to 10 tons per
hectare from well-managed plantations, have been reported
recently on the USDA-FAS website. Terefore, it would be
economically intuitive to consider palm oil as a favourable
feed stock for biodiesel production. On the other hand, Shah
and Gupta
38
argued that it is more reasonable to use ined-
ible oils such as Jatropha oil, as edible oils are not in surplus
supply. Despite this, the majority of reported studies used
soybean oil
26, 27, 31, 34, 44, 45
and sunfower oil.
41, 47, 48
In addition,
some work has been reported on the enzymatic production
of biodiesel using olive oil,
30
rice bran oil
29
and canola oil.
28

A comparison between the production yields of various
vegetable oils and the physical and chemical properties of
biodiesel fuels produced from them are shown in Table 3.
49, 50
Other feedstock
It has been reported that 6090% of the biodiesel cost arises
from the cost of the feedstock oil.
29
Tus, the use of waste
cooking oil should greatly reduce the cost of biodiesel.
Table 3. Comparison between the physical and chemical properties of biodiesel fuels produced from various
vegetable oils.
Vegetable Production Kinematic Cetane

Lower

Cloud Flash

Density
oil used yield viscosity number
48
heating value point point

(g L
-1
)
48

(kg ha
1
)
49
(mm
2
s
-1
)
48
(MJ L
-1
)
48
(
o
C)
48
(
o
C)
48
Peanut 890 4.9 (37.8
o
C) 54 33.6 5 176 0.883
Soybean 375 4.5 (37.8
o
C) 45 33.5 1 178 0.885
Soybean 375 4.0 (40
o
C) 45.756 32.7 0.880 (15
o
C)
Babassu - 3.6 (37.8
o
C) 63 31.8 4 127 0.879
Palm 4000 5.7 (37.8
o
C) 62 33.5 13 164 0.880
Palm 4000 4.34.5 (40
o
C) 64.370 32.4 0.872-0.877 (15
o
C)
Sunflower 655 4.6 (37.8
o
C) 49 33.5 1 183 0.860
Rapeseed 1000 4.2 (40
o
C) 5 l59.7 32.8 0.882 (15
o
C)
Used rapeseed - 9.48 (30
o
C) 53 36.7 192 0.895
Used corn oil - 6.23 (30
o
C) 63.9 42.3 166 0.884
62 2007 Society of Chemical Industry and John Wiley & Sons, Ltd | Biofuels, Bioprod. Bioref. 1:5766 (2007); DOI: 10.1002/bbb
S Al-Zuhair Review: Production of biodiesel: possibilities and challenges
In addition, production of biodiesel from waste edible
oil is considered an important step in reducing and recy-
cling waste oil. A fresh vegetable oil and its waste difer
signifcantly in water (around 2000 ppm) and free fatty acid
(1015%) contents.
19, 29
Terefore, alkaline catalyst cannot
be utilized, and chemical catalysts are limited to the slower
acidic ones.
19
Indeed, the use of waste oil as a feedstock for
biodiesel production by acidic catalyst has been reported
in the literature.
19, 51, 52
On the other hand, lipase is capable
of converting all the free fatty acids contained in waste oils
to esters.
30
It has been reported that the reaction proceeds
in a similar manner when using Novozym 435 to produce
biodiesel from virgin and used olive oils, and the conver-
sion was almost identical for both cases, which confrms the
applicability of lipase with waste cooking oil.
30
In addition,
up to 500 ppm water found in waste vegetable oil decreased
the rate of methanolysis but did not afect the equilibrium of
the reaction.
33

On the other hand, animal fats have been used for the
production of biodiesel.
53
However, due to the high melting
points, which are usually near the denaturation temperature
of lipase, the reaction has to take place in an organic solvent
media to dissolve the solid fat. Te addition of organic
solvent is not recommended, as it requires the addition of a
solvent recovery unit. Other sources of oils have also been
evaluated as raw material for biodiesel production, including
waste oil adsorbed on activated bleaching earth (ABE)
35
and
cellular biomass of oleaginous yeasts and flaments fungi.
54
Acyl-acceptors
Alcohols
Methanol is the most commonly used alcohol in biodiesel
production, mainly because of its reactivity, as shown
in Table 2. Although methanol is easier to produce than
ethanol, sustainable methods of methanol production are
currently not economically viable and the majority of it is
formed from syngas, which is extracted from natural gas
(a non-renewable source). In addition, methanol is the most
toxic and has the most deleterious efect on the biocatalyst
activity in comparison to other alcohols. On the other hand,
ethanol can easily be formed from renewable sources by
fermentation, which makes the process of biodiesel
production, totally green. Terefore, ethanol is more readily
accepted for use in a variety of industrial situations.
Te efect of methanol on the enzymatic production
of biodiesel has been thoroughly discussed in the litera-
ture. Shimada et al.
42
reported that biodiesel production
increased with increasing methanol concentration up to
an oil-to-methanol ratio of 3:1, and then decreased when
methanol concentration was further increased. Tis was also
found by Noureddini et al.
34
although the ratio was higher
(7.5:1). In general, it is widely accepted that methanol that
is completely dissolved in the substrate mixture should not
inactivate the lipases.
33, 39, 42
Proteins generally are unstable in
short-chain alcohols such as methanol and ethanol. Tere-
fore, lipases are inactivated by contact with insoluble meth-
anol, which exists as drops in the oil. To avoid this problem
the amount of methanol used should be below its solubility
limits in oil. To achieve this ef ciently, stepwise addition of
methanol in a way to keep the amount below its solubility
has been proposed.
33, 42
Further, lipases having substantial
methanol resistance, such as those extracted from Pseu-
domonas strains or using long-chain fatty alcohols that do
not inhibit the enzyme could be used to overcome methanol
inhibition problems. In addition, the use of organic solvent
to dissolve the methanol has also been suggested; however,
this is not recommended as it requires the addition of a
solvent recovery unit. On the other hand, methyl acetate has
been proposed to replace alcohols as an acyl acceptor for
biodiesel production from soybean oil using Novozym 435.
26

It was shown that methyl acetate had no negative efect on
enzymatic activity and almost no loss in lipase activity was
detected even afer being continuously used for 100 batches.
However, when methanol was used in an appropriate
concentration, its reaction rate remained much faster.
Immobilization of lipase
Lipases are spontaneously soluble in aqueous solutions as
a result of their globular protein nature
55
but their natural
substrates (i.e. lipids) are not. Using a suitable organic
solvent or an emulsifer helps to overcome the problem
of intimate contact between substrate and enzyme.
25, 56

However, the practical use of lipase in such psuedohomoge-
neous reaction systems poses technological dif culties such
63
Review: Production of biodiesel: possibilities and challenges S Al-Zuhair
2007 Society of Chemical Industry and John Wiley & Sons, Ltd | Biofuels, Bioprod. Bioref. 1:5766 (2007); DOI: 10.1002/bbb
as contamination of the products with residual enzymatic
activity, and economic dif culties such as the use of enzyme
in a single reactor pass. Hence, part of the overall potential
enzymatic activity is lost. If the lipase is immobilized, then
it becomes an independent phase within the reaction system,
which may easily be retained in the reactor with concomi-
tant advantages in preventing contamination of the products
and extending its useful active life. Further, increasing the
temperature generally increases the rate of lipase-cata-
lyzed reaction per unit amount of active enzyme; however,
increasing the temperature also leads to a higher thermal
deactivation rate of the lipase itself, thus yielding decreasing
amounts of active enzyme.
25, 57
Because immobilization
provides a more rigid external backbone for lipase molecule,
temperature optima are expected to increase,
56
which results
in a faster reaction rate.
Among all the immobilization methods, physical adsorp-
tion has been clearly selected by most researchers due to
its ease, absence of expensive and toxic chemicals, ability
to retain the activity, and feasibility of regeneration.
58

On the other hand, poor adsorption of enzyme results in
its leaching of the support surface, which favours other
means of enzyme immobilization such as covalent bonding,
entrapment and encapsulation. It is possible to strengthen
the attachment between the water-soluble enzyme and the
water-insoluble surfaces by using multifunctional agents
that are bifunctional in nature and have low molecular
weight, such as glutaraldehyde.
55, 59, 60
Te author of this
review has studied the kinetics of lipase adsorption and
a temperature dependence correlation for the amount of
adsorbed lipase activity and that present in the supernatant
solution was determined.
60
Immobilized lipase by physical
adsorption has been successfully used in biodiesel produc-
tion.
27, 36, 37, 4246
On the other hand, lipase immobilized by
entrapment is much more stable than physically adsorbed
lipase, and unlike the covalent bonding,
60
this method uses
a relatively simple procedure. Hsu et al.
61
developed a novel
procedure for the immobilization of lipase from B. cepacia
within a phyllosilicate solgel matrix. Te applicability of
using lipase encapsulated in a solgel in biodiesel produc-
tion has shown encouraging results.
34, 62, 63
Dispersing immobilized enzyme in a stirred tank reactor
promotes intimate contact of the enzyme with the reaction
mixture. Te immobilized enzyme is retained within
the reactor by means of a flter at the outlet. Low costs of
construction are normally associated with these reactors.
However, as the reaction proceeds, glycerol is generated
and physically mixes with the alcohol to form a second
liquid phase that is not completely miscible with the oil.
Tis second polar organic phase serves to extract alcohol
from the oil phase, thereby decreasing the concentra-
tion of this substrate in the reaction medium and causing
a concomitant decrease in the conversion achieved in a
fxed amount of time. Consequently, conversions will be
enhanced if glycerol is removed from the substrate mixture
as the reaction proceeds.
46
To achieve this, membrane reac-
tors with immobilized lipase are proposed, which may take
either a fat sheet
64
or hollow fbre form.
37, 65
In this type of
reactor, continuous reaction and separation can take place
simultaneously.
Conclusions
Biodiesel has very similar properties to petroleum diesel but
with much lower sulfur, CO
2
and particulate emissions. In
conventional chemical processing, synthesis of fatty acid
esters (biodiesel) is achieved by alkaline- or acidic-catalyzed
transesterifcation of triacylglycerol (oil). Te acid-catalyzed
production of biodiesel has received less attention because
it is 400 times slower than production using alkali catalyst.
Nevertheless, the acid-catalyzed approach is less sensitive to
free fatty acids in the feedstock oil than the alkali-catalyzed
process. Transesterifcation by either chemical catalyst has
several drawbacks, which include being energy intensive,
dif cult to recover glycerol and requires wastewater treat-
ment. In addition, free fatty acids present in the oil interfere
with the reaction, especially for the alkaline catalyst case,
leading to undesirable side products. A less energy inten-
sive and environmental friendly procedure would be to use
enzymes for oil transesterifcation. Enzymatic transesteri-
fcation can overcome the problems facing conventional
chemical methods without compromising the advantages of
using alkaline- catalyzed reactions. Most importantly, glyc-
erol can be easily recovered without any complex process
and free fatty acids contained in the oils can be completely
converted to methyl esters. In addition, with enzymatic
64 2007 Society of Chemical Industry and John Wiley & Sons, Ltd | Biofuels, Bioprod. Bioref. 1:5766 (2007); DOI: 10.1002/bbb
S Al-Zuhair Review: Production of biodiesel: possibilities and challenges
catalyzed reactions, subsequent wastewater treatment is
not required. Production of biodiesel from waste oil greatly
reduces the cost of biodiesel and is considered an important
step in reducing and recycling waste oil. Methanol is the
most commonly used alcohol in biodiesel production due
to its low cost and high reactivity. However, methanol has a
much more deleterious efect on the biocatalyst activity than
ethanol. Terefore, by using ethanol the reusability of the
biocatalyst is enhanced. In addition, ethanol can be easily
formed from renewable sources by fermentation, which
makes the process of biodiesel production totally green.
Tere are two main problems facing the efective utiliza-
tion of lipase in the production of biodiesel. First, lipases are
unstable in short-chain alcohols. To avoid this problem, the
amount of alcohol used must be less than the solubility limit,
so that it is not present as a separate phase. Stepwise addition
of alcohol has been proposed as a way of keeping the amount
below the critical concentration. Te use of an organic
solvent has also been proposed as a way of increasing the
alcohol concentration. In addition, it is recommended to
use lipases with substantial ethanol resistance, such as those
extracted from Pseudomonas strains. Second, the presence
of water is needed for the expression of enzyme activity, but
results in lipase inactivation. Nevertheless, R. meihei lipases
have ef ciently catalyzed alcoholysis of oil even in the pres-
ence of up to 20% water.
Te cost of lipase remains the main obstacle facing full
exploitation of its potential. Terefore, the reuse of lipase
is essential from the economic point of view, which can be
achieved by using the lipase in immobilized form. In addi-
tion, immobilized lipase displays improved stability and
activity in comparison to its use in soluble form. Immobi-
lization of the lipase can be achieved by encapsulation in
solgel or by attachment to a solid surface.
With the inevitable depletion of the non-renewable
resources of fossil fuels, and due to its favourable envi-
ronmental features, biodiesel promises to be the fuel of
tomorrow. Nevertheless, there are many obstacles standing
in the way of the efective production of biodiesel on an
industrial scale. Te most important challenges and the
proposed ways to overcome them have been presented in
this review. At a laboratory scale, these solutions have shown
very promising results.
References
1. Korbitz W, Biodiesel production in Europe and North America. An
encouraging prospect. Renewable Energy 16:10781083 (1999).
2. Agarwal AK and Das LM, Biodiesel development and characterization for
use as a fuel in compression ignition engines. J Eng Gas Turbines Power
123:440447 (2001).
3. Krawcsyk T, Biodiesel: alternative fuel makes inroads but hurdles remain.
Inform 7:800815 (1996).
4. Von Wedel R, Technical handbook for marine biodiesel in recreational
boats. Prepared for National Renewable Energy Laboratory, US
Department of Energy, Subcontract No. ACG- 7-16688-01 under Prime
Contract No. DE-AC36-83CH10093, pp 32 (1999).
5. Yusaf T, Al-Zuhair S and Al-Atabi M, Performance of diesel engine using
an emulsion of biodieselconventional diesel fuel. J Mech Eng 56:
137142 (2005).
6. Connemann J and Fischer J, Biodiesel in Europe 1998: biodiesel
processing technologies. Paper presented at the International Liquid
Biofuels Congress, Brazil (1998).
7. Freedman B, Buttereld RO and Pryde EH, Transesterication kinetics of
soybean oil. J Am Oil Chem Soc 63:13751380 (1986).
8. Noureddini H and Zhu D, Kinetics of transesterication of soybean oil.
J Am Oil Chem Soc 74:14571463 (1997).
9. Antoln G, Tinaut FV, Briceno Y, Castano V, Perez C and Ramrez AI.
Optimisation of biodiesel production by sunower oil transesterication
Bioresource Technol 83:111114 (2002).
10. Darnoko D and Cheryan M, Kinetics of palm oil transesterication in a
batch reactor. J Am Oil Chem Soc 77:12631267 (2000).
11. Demirbas A, Biodiesel production from vegetable oils via catalytic and
non-catalytic supercritical methanol transesterication methods. Prog
Energy Comb. Sci. 31:466487 (2005).
12 Kreutzer UR, Manufacture of fatty alcohols based on natural fats and oils.
J Am Oil Chem Soc 61:343348 (1984).
13. Liu KS, Preparation of fatty acid methyl esters for gaschromatographic
analysis of lipids in biological materials. J Am Oil Chem Soc
71:11791187 (1994).
14. Basu HN and Norris ME, Process for production of esters for use as a
diesel fuel substitute using a non-alkaline catalyst. US Patent 5525126
(1996).
15. Jeromin L, Peukert E and Wollmann G, Process for the pre-esterication
of free fatty acids in fats and oils. US Patent 4698186 (1987).
16. Lepper H and Friesenhagen L, Process for the production of fatty acid
esters of short-chain aliphatic alcohols from fats and/or oils containing
free fatty acids. US Patent 4608202 (1986).
65
Review: Production of biodiesel: possibilities and challenges S Al-Zuhair
2007 Society of Chemical Industry and John Wiley & Sons, Ltd | Biofuels, Bioprod. Bioref. 1:5766 (2007); DOI: 10.1002/bbb
17. Watanabe Y, Shimada Y, Sugihara A and Tominaga Y, Enzymatic
conversion of waste edible oil to biodiesel fuel in a xedbed bioreactor.
J Am Oil Chem Soc 78:703707 (2001).
18. Canakci M and Van Gerpen J, Biodiesel production from oils and fats
with high free fatty acids. Trans ASAE 44:142936 (2001).
19. Zhang Y, Dube MA, McLean DD and Kates M, Biodiesel production from
waste cooking oil: 1. Process design and technological assessment
Bioresource Technol 89:116 (2003).
20. Demirbas A, Biodiesel from vegetable oils via transesterication in
supercritical methanol. Energy Convers Manage 43:23492356
(2002).
21 Janssen AEM, Vaidya AM and Halling PJ, Substrate specicity and
kinetics of C. rugosa lipase in organic media. Enzyme Microbiol Technol
18:340346 (1996).
22. Verger R, Maria CE and DeHaas GH, Action of phospholipase A at
interfaces. J Biological Chem 218:40284034 (1973).
23. Panalotov I and Verger R, Physical Chemistry of Biological Interfaces.
Marcel Dekker Inc., New York (2000).
24. Rooney D and Weartherley LR, The effect of reaction conditions upon
lipase catalyzed hydrolysis of high oleate sunower oil in a stirred
liquid-liquid reactor. Process Biochem 36:947953 (2001).
25. Al-Zuhair S, Hasan M and Ramachandran KB, Kinetic hydrolysis of palm
oil using lipase. Proc Biochem 38:11551163 (2003).
26. Wei D, Yuanyuan X, Dehua L and Jing Z, Comparative study on lipase-
catalyzed transformation of soybean oil for biodiesel production with
different acyl acceptors. J Mol Catal B: Enzymatic 30:125129 (2004).
27. Samukawa T, Kaieda M, Matsumoto T, Ban K, Kondo A, Shimada Y,
Noda H and Fukuda H, Pretreatment of immobilized Candida antarctica
lipase for biodiesel fuel production from plant oil. J Biosci Bioeng
90:180183 (2000).
28. Chang HM, Liao HF, Lee CC and Shieh CJ, Optimised synthesis of
lipase-catalyzed biodiesel by Novozym 435. J Chem Technol Biotechnol
80:307312 (2005).
29. Lai CC, Zullaikah S, Vali SR and Ju YH. Lipase-catalyzed production
of biodiesel from rice bran oil. J Chem Technol Biotechnol 80:331337
(2005).
30. Sanchez F and Vasudevan PT, Enzyme catalyzed production of biodiesel
from olive oil. Appl Biochem Biotechnol 135:114 (2006).
31. Kaieda M, Samukawa T, Matsumoto T, Ban K, Kondo A, Shimada Y,
Noda H, Nomotos F, Ohtsuka K, Izumoto E and Fukuda H, Biodiesel fuel
production from plant oil catalyzed by Rhizopus oryzae lipase in a
water-containing system without an organic solvent. J Biosci Bioeng
88:627631 (1999).
32. Kaieda M, Samukawa T, Kondo A and Fukuda H, Effect of methanol and
water contents on production of biodiesel fuel from plant oil catalyzed by
various lipases in a solvent-free system. J Biosci Bioeng 91:1215
(2001).
33. Shimada Y, Watanabe Y, Sugihara A and Tominaga Y, Enzymatic
alcoholysis for biodiesel fuel production and application of the reaction
to oil processing. J Mol Catal B: Enzymatic 17:133142 (2002).
34. Noureddini H, Gao X and Philkana RS, Immobilized Pseudomonas
cepacia lipase for biodiesel fuel production from soybean oil.
Bioresource Technol 96:769777 (2005).
35. Lara PV and Park EY, Potential application of waste activated bleaching
earth on the production of fatty acid alkyl esters using Candida
cylindracea lipase in organic solvent system Enz Microb Technol
34:270277 (2004).
36. Hilal N, Kochkodan V, Nigmatullin R, Goncharuk V and Al-Khatib L,
Lipase-immobilized biocatalytic membranes for enzymatic esterication:
Comparison of various approaches to membrane preparation.
J Membrane Sci 268:198207 (2006).
37. Hilal N, Nigmatullin R and Alpatova A, Immobilization of crosslinked
lipase aggregates within microporous polymeric membranes.
J Membrane Sci 238:131 (2004).
38. Shah S and Gupta MN, Lipase catalyzed preparation of biodiesel from
Jatropha oil in a solvent free system. Proc Biochem 42:409413
(2006).
39. Al-Zuhair S, Jayaraman KV, Krishnan S and Chan YH, The effect of fatty
acid concentration and water content on the production of biodiesel by
lipase. Biochem Eng J 30:212217 (2006).
40. Iso M, Chen B, Eguchi M, Kudo T and Shrestha S. Production of biodiesel
fuel from triglycerides and alcohol using immobilized lipase. J Mol Catal
B: Enzymatic 16:5358 (2001).
41. Hama S, Yamaji H, Kaieda M, Oda M, Kondo A and Fukuda H, Effect
of fatty acid membrane composition on whole-cell biocatalysts for
biodiesel-fuel production. Biochem Eng J 21:155160 (2004).
42. Shimada Y, Watanabe Y, Samukawa T, Sugihara A, Noda H, Fukuda H
and Tominaga Y, Conversion of vegetable oil biodiesel using immobilized
Candida antarctica lipase. J Am Oil Chem Soc 76:789793 (1999).
43. Tweddell RJ, Kermasha S, Combes D and Marty A, Esterication and
interesterication activities of lipase from Rhizopus niveus and Mucor
miehei in three different types of organic media: a comparitive study.
Ezyme Microb Technol 22:439445 (1998).
44 Watanabe Y, Shimada Y, Sugihara A, Noda H, Fukuda H and Tominaga
Y, Continuous production of biodiesel fuel from vegetable oil using
immobilized Candida antarctica lipase. JAOCS 77:355360 (2000).
66 2007 Society of Chemical Industry and John Wiley & Sons, Ltd | Biofuels, Bioprod. Bioref. 1:5766 (2007); DOI: 10.1002/bbb
S Al-Zuhair Review: Production of biodiesel: possibilities and challenges
45. Watanabe Y, Shimada Y, Sugihara A and Tominaga Y, Conversion
of degummed soybean oil to biodiesel fuel with immobilized Candida
antarctica lipase. J Mol Catal B: Enzymatic 17:151155 (2002).
46. Watanabe Y, Shimada Y, Sugihara A and Tominaga Y, Stepwise
ethanolysis of tuna oil using immobilized Candida antarctica lipase.
J Biosci Bioeng 88:622626 (1999).
47. Orcaire O, Buisson P and Pierre AC, Application of silica aerogel
encapsulated lipases in the synthesis of biodiesel by transesterication
reactions. J Mol Catal B: Enzymatic 42:106113 (2006).
48. Soumanou MM and Bornscheuer UT, Improvement in lipase-catalyzed
synthesis of fatty acid methyl esters from sunower oil. Enzyme Microbiol
Technol 33:97103 (2003).
49 Fukuda H, Kondo A and Noda H, Biodiesel fuel production by
transesterication of oils. J Biosci Bioeng 92:405416 (2001).
50. http://journeytoforever.org/biodiesel_yield.html#alph
51. Al-Widyan MI and Al-Shyoukh AO, Experimental evaluation of the
transesterication of waste palm oil into biodiesel. Bioresource
Technol 85:253256 (2002).
52. Al-Widyan MI, Tashtoush G and Abu-Qudais M, Utilization of ethyl ester
of waste vegetable oils as fuel in diesel engines. Fuel Process Technol
76:91103 (2002).
53. Tashtoush GM, Al-Widyan MI and Al-Jarrah MM, Experimental study
on evaluation and optimization of conversion of waste animal fat into
biodiesel. Energy Convers Manag 45:2697,2711 (2004).
54. Miao XL and Wu Q, Biodiesel production from heterotrophic microalgal
oil. Bioresource Technol 97:841846 (2006).
55. Bjorkling F, Godtfredsen SE and Kirk O, The future impact of industrial
lipases. Trends Biotechnol 9:360363 (1991).
56. Knezevic ZD, Siler-Marinkovic SS and Mojovic LV, Kinetics of lipase-
catalyzed hydrolysis of Palm oil in lecinthin/isooctane reversed micelles.
Appl Microbiol Biotechnol 49:267271 (1998).
57. Shamel MM, Ramachandran KB and Hasan M, Effect of hollow ber
membrane material and operating conditions on lipase adsorption.
IMSTEC 03, The Fifth International Membrane Science & Technology
Conference, Australia (2003).
58 Guit RPM, Kloosterman M, Meindersma GW, Mayer M and Meijer
EM, Lipase kinetics: hydrolysis of triacetin by lipase from Candida
cylindracea in a hollow bre membrane reactor. Biochem Bioeng
38:727732 (1991).
59. Balcao VM, Paiva AL and Malcata FX, Bioreactors with immobilized
lipases: state of the art. Enz Microb Technol 18:392416 (1996).
60. Shamel MM, Ramachandran KB, Hasan M and Al-Zuhair S. Hydrolysis
of palm and olive oils by immobilized lipase using hollow-bre reactor.
Biochem Eng J 34:228235 (2007).
61. Hsu AF, Jones K, Marmer WN and Foglia TA, Production of alkyl esters
from tallow and grease using lipase immobilized in a phyllosilicate
solgel. J Am Oil Chem Soc 78(6):585588 (2001).
62. Chen JP and Lin YS, Solgel powders and supported solgel polymers
for immobilization of lipase in ester synthesis. Enz Microb Technol
32:801811 (2003).
63. Vidinha P, Augusto V, Almeida M, Fonseca I, Fidalgo A, Ilharco L,
Joaquim MSC and Barreiros S, Solgel encapsulation: an efcient and
versatile immobilization technique for cutinase in non-aqueous media.
Biotechnol 121:2333 (2006).
64. Isono Y, Nakajima M and Nabetani H, Solvent-free esterication of oleic
acid and oleyl alcohol using membrane reactor and lipase-surfactant
complex. J Ferment Bioeng 86:138140 (1998).
65. Shamel MM, Al-Zuhair S and Azaha R, Adsorption of lipase on hollow
bre membrane chips. Int J Art Cells, Blood SubstitutesBiotechnol
33:423433 (2005).