Biotechnology Advances 29 (2011) 124–141

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Biotechnology Advances
j o u r n a l h o m e p a g e : w w w. e l s ev i e r. c o m / l o c a t e / b i o t e c h a d v

Research review paper

Renewable and sustainable bioenergies production from palm oil mill effluent
(POME): Win–win strategies toward better environmental protection
Man Kee Lam, Keat Teong Lee ⁎
School of Chemical Engineering, Universiti Sains Malaysia, Engineering Campus, Seri Ampangan, 14300 Nibong Tebal, Pulau Pinang, Malaysia

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

Article history: Palm oil industry is one of the leading agricultural industries in Malaysia with average crude palm oil
Received 28 July 2010 production of more than 13 million tonne per year. However, production of such huge amount of crude palm
Received in revised form 2 October 2010 oil has consequently resulted to even larger amount of palm oil mill effluent (POME). POME is a highly
Accepted 3 October 2010
polluting wastewater with high chemical oxygen demand (COD) and biochemical oxygen demand (BOD) in
Available online 19 October 2010
which can caused severe pollution to the environment, typically pollution to water resources. On the other
Keywords:
hand, POME was identified as a potential source to generate renewable bioenergies such as biomethane and
Palm oil mill effluent (POME) biohydrogen through anaerobic digestion. In other words, a combination of wastewater treatment and
Biomethane renewable bioenergies production would be an added advantage to the palm oil industry. In line with the
Biohydrogen world's focus on sustainability concept, such strategy should be implemented immediately to ensure palm oil
Biodiesel is produced in an environmental friendly and sustainable manner. This review aims to discuss various
Bioethanol technologies to convert POME to biomethane and biohydrogen in a commercial scale. Furthermore, discussion
Microalgae on using POME to culture microalgae for biodiesel and bioethanol production was included in the present
paper as a new remedy to utilize POME with a greater beneficial return.
© 2010 Elsevier Inc. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 125
2. POME . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 125
3. Palm oil mill process flow description . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 125
3.1. Sterilization of fresh fruit bunches . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 125
3.2. Stripping, digestion and pressing of fruits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126
3.3. Clarification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126
3.4. Kernel oil recovery . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126
4. Bioenergies production from POME . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126
4.1. Biomethane production . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126
4.1.1. Biochemistry of anaerobic digestion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 127
4.1.2. Conventional ponding (lagoon) system . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128
4.1.3. Open digesting tanks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 129
4.1.4. Closed digesting tanks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 129
4.1.5. Other related technologies and future direction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 130
4.2. Biohydrogen production . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131
4.3. POME cultured microalgae: a novel approach to produce biofuels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133
4.3.1. Microalgae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133
4.3.2. POME as nutrients source to culture microalgae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 134
4.3.3. Overall system design . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 135
4.3.4. Open pond and closed photo-bioreactor system . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 135
4.3.5. Mitigation of CO2 from palm oil mill by microalgae cultivation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 136
4.3.6. Microalgae biomass harvesting method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 136
4.3.7. Biodiesel production from microalgae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 137
4.3.8. Bioethanol production from microalgae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 139

⁎ Corresponding author. Tel.: +60 4 5996467; fax: +60 4 5941013.

E-mail address: chktlee@eng.usm.my (K.T. Lee).

0734-9750/$ – see front matter © 2010 Elsevier Inc. All rights reserved.
doi:10.1016/j.biotechadv.2010.10.001
 M.K. Lam, K.T. Lee / Biotechnology Advances 29 (2011) 124–141 125

5. Conclusion. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 139
Acknowledgement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 139
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 139

1. Introduction
The Malaysian palm oil industry has grown rapidly over the past few
years to become the world's second largest producer of palm oil,
accounted for 10.3% of the world's oils and fats production in year 2007
(Lam et al., 2009c). The industry served as an important backbone to the
country's economy and has significantly increased the standard of living of
its populations (Yusoff and Hansen, 2007). For example, in year 2008, the
total export of palm oil and derived products raked RM 64,808 million
(USD 20,268 million), or 9.8% of the total national revenue (Yusof and
Yew, 2009). Furthermore, the palm oil industry provides a source of
livelihood to rural families in government land schemes and private small
holders, as well as employment opportunities to agricultural workers in
estates (Wu et al., 2010). Currently, palm oil is even intensively used as a
source to produce biodiesel after the announcement of Fifth Fuel Policy
under Eighth Malaysia Plan (2001–2005) (Lim and Teong, 2010). The new
policy is drafted with the mission to lead Malaysia towards green energy
utilization and palm oil has re-position itself as an important asset to drive
this policy forward.
However, sustainability of palm oil production has always been
questioned, especially by non-governmental organizations (NGOs). It was
claimed that further expansion of oil palm industry will cause severe
negative impacts toward environment such as deforestation, destruction
of orangutan habitat and serious greenhouse gases (GHG) emission due to
over exploration of peat land for oil palm plantation (Laurance et al., 2010;
Yule, 2010). Nevertheless, criticisms are not only limited to plantation
sector, but also include palm oil mills. The most significant pollutant from
palm oil mills is POME (Poh and Chong, 2009). If there is no proper waste
management practices implemented in palm oil mills, POME can cause
water and air pollution (biomethane emission). This problem become
more challenging as the number of palm oil mills in Malaysia has
increased tremendously in the past few years, from about 10 mills in 1960
to 410 operating mills in 2008 (Wu et al., 2010). Hence, immediate
remedies should be taken to overcome this problem and to further
strengthen the palm oil sustainability concepts announced by Malaysian
Palm Oil Board (MPOB) and Malaysian Palm Oil Council (MPOC).
The purpose of the present paper depicts some potential strategies to
reduce the environmental problems created by POME coupled with
renewable energies generation, such as biomethane and biohydrogen. In
addition, the prospects and technologies to commercialize those
renewable energies are discussed in details. Apart from that, using
POME to culture microalgae for biodiesel and bioethanol production
combined with wastewater treatments were proposed as a new method
to enhance renewable energies production from palm oil mills. This
visionary strategy will eventually become an important baseline reference
for governments, researchers and palm oil millers.
mills base on the total crude palm oil production of 17.56 million tonnes
(Malaysian Palm Oil Board, 2010). In fact, the palm oil industry was
identified as one of the agricultural industry in Malaysia that generates the
highest pollution load into rivers throughout the country (Wu et al.,
2007). Although POME is a non-toxic liquid waste with unpleasant smell,
its COD and BOD value are high enough to cause serious pollution and
environmental problem to the rivers. Table 1 shows a typical character-
istics of POME (Malaysian Palm Oil Board) and discharge limits for POME
discharge into water sources in Malaysia. Nevertheless, many palm oil
mills are still unable to adhere to the wastewater discharge limits and thus
resulting to a dramatic increase in the number of polluted rivers (Ahmad
and Chan, 2009). Furthermore, if the POME is discharged without prior
purification, the BOD produced in year 2009 was estimated to be
1095 million kg. In other words, if each human being is assumed to
produce 14.6 kg annually, this BOD value will be equivalent to the waste
generated by 75 million people which is nearly thrice the current
population in Malaysia (Ahmad and Chan, 2009).
3. Palm oil mill process flow description
The wet palm oil milling process is the most common way of
extracting palm oil from fresh fruit bunches (FFB), typically in Malaysia
(Wu et al., 2010). It involves several stages in which huge amount of water
and steam are required for washing and sterilizing. Thus, this has resulted
to huge amount of wastewater generated from palm oil mill or better
known as POME. Fig. 1 shows a simplified process flow diagram to
produce palm oil and the description of some important stages is
explained in the following section.
3.1. Sterilization of fresh fruit bunches
During sterilization process, pressurized steam (3×105 Pa) at high
temperature (140 °C) (Wu et al., 2010) is used to infuse moisture into the
nuts that can cause it to expand. Consequently, the contraction of the nut
will cause the detachment of the kernel from the shell wall and resulting
to loosen kernels within their shells (Poku, 2002). Apart from that, another
purpose of sterilization is to destroy the oil-splitting enzymes and thus
slows down the formation of free fatty acids in the oil. However, it is
important to ensure that air is evacuated from sterilizer in order to
increase the efficiency of heat transfer and to avoid oil oxidation by air
Table 1
Characteristics of POME and discharge limits.
Parametersa POME (range) POME (mean) Discharge standard
(1-1-1984 and thereafter)

2. POME Temperature (°C) 80–90 85 45

pH 3.4–5.2 4.2 5.0–9.0
Oil and grease 130–18,000 6000 50
Raw POME is a colloidal suspension containing 95–96% water, 0.6–
BOD3b 10,250–43,750 25,000 100
0.7% oil and 4–5% total solids including 2–4% suspended solids. The COD 15,000–100,000 51,000 –
suspended solids are mainly consist of debris from palm fruit mesocarp Total solid 11,500–79,000 40,000 –
generated from three main sources, (1) sterilizer condensate, (2) sludge Suspended solid 5000–54,000 18,000 400
Total volatile solid 9000–72,000 34,000 –
separator and (3) hydrocyclone waste (Borja and Bnaks, 1994; Khalid and
Total nitrogen 180–1400 750 200c
Wan Mustafa, 1992). Consequently, about 0.9, 1.5 and 0.1 m3 of POME will Ammoniacal 4–80 35 150c
be generated from sterilizer condensate, sludge separator and hydro- nitrogen
cyclone waste for each tonne (1.13 m3) of crude palm oil processed (Ma a
All parameters are in mg L− 1 except temperature and pH.
and Ong, 1988). Thus, it was estimated that in year 2009, 43.8 million m3 b
Sample is incubated for 3 days at 30 °C.
(11,600 million gallon) of POME was generated from Malaysian palm oil c
Value of filtered sample.
126 M.K. Lam, K.T. Lee / Biotechnology Advances 29 (2011) 124–141
Fig. 1. Flow diagram of a palm oil mill.
(Poku, 2002). The condensate coming out from sterilizer constitutes as
one of the major sources of POME.
3.2. Stripping, digestion and pressing of fruits
The purpose of stripping is to separate the sterilized fruits from the
bunch stalks by using a rotary drum thresher (Wu et al., 2010). The
detached fruits are passed through a bar screen in the stripper,
collected by a bucket conveyor and then channeled into a digester. In
the digester, the fruit is softened or mashed by steam-heated
cylindrical vessel fitted with a central rotating shaft carrying a
number of mechanical arms. Through the action of rotation at high
temperature (80–90 °C) (Wu et al., 2010), the fruits' outer covering
(mesocarp) are loosened from the nuts and subsequently being
channeled to a mechanical press machine to squeeze out the crude
palm oil.
3.3. Clarification
The purpose of clarification is to separate the oil from its entrained
impurities. The fluid flowing out from the press machine is a mixture
of palm oil, water, cell debris, fibrous material and other insoluble
solids that have high viscosity. Hence, hot water is added into the
clarifier to break the oil emulsion and acts as a barrier to cause the
insoluble solids to settle to the bottom of the clarifier while the lighter
oil droplets flow through the watery mixture on the top. Consequent-
ly, the bottom phase from the clarifier is drained off as sludge or POME
for further purification before being discharged.
3.4. Kernel oil recovery
Residue (press cake) from the press machine consists of a mixture of
fibre and nuts. After the separation of fibre from nuts by strong air current
induced by a suction fan, the fibre is usually sent to boiler house as boiler
fuel while the nuts are sent to a nut cracker and finally to hydrocyclone.
Hydrocyclone is commonly used to separate the kernels from empty shells
after cracking the nuts (Wu et al., 2010). The discharge from this process
constitutes the last source of POME.
4. Bioenergies production from POME
Releasing un-treated POME directly to the surrounding will caused
serious pollution problem towards the environment. Instead, POME can
be treated efficiently using available wastewater treatment technologies
in order to meet the standard discharge limits for wastewater. However, in
conjunction with the need to address global sustainability challenges,
POME treatment has now evolved into another new dimension with
simultaneous bioenergies recovery strategy. The main benefit of
implementing this approach is reduction in wastewater treatment cost
by producing green energies as by-products that is also very beneficial
towards environmental protection. The following sections will describe
some of the technologies capable to recover biomethane and biohydrogen
from POME. Apart from that, a conceptual idea of utilizing POME as a low
cost nutrient source to cultivate microalgae for biodiesel and bioethanol
production will also be presented.
4.1. Biomethane production
Biological treatment in wastewater processes has been accepted as an
effective way to remove dissolved and biodegradable constituents by
utilizing variety of microorganisms, principally bacteria (Sankaran et al.,
2010). Since POME contains high level of organic matters and thus,
adoption of anaerobic digestion in the first stage of the treatment process
is a necessity to convert the bulk of the wastes to biogas (biomethane). The
treated effluent is further subjected to an aerobic treatment in order to
meet the required discharge standards. These treatment steps have been
 M.K. Lam, K.T. Lee / Biotechnology Advances 29 (2011) 124–141 127

applied either as an open pond or open digesting tank systems in Table 3

Malaysian palm oil mills. However, due to the lack of infrastructure and Methane emission rate from anaerobic digestion of POME.

low demand of renewable energy (biogas) in the county, biomethane is No Reactor type Conditions HRT Methane COD Reference
not captured but escapes directly to the atmosphere and thus caused (days) emission removal
serious air pollution. Methane has been categorized as one of the GHG rate (L efficiency
methane/g
with its global warming potential 21 times more potent than CO2. A life
COD
cycle assessment study on Malaysian palm oil mill processes revealed that removed)
the non-recovered biomethane emission from POME contributed the
1 Single stage – 6.2 0.325 96% Borja and
highest impact towards the environment (climate change category) and stirred Banks, 1994
therefore makes the overall processes not environmentally friendly digester
(Subramaniam et al., 2008). Table 2 shows an estimation of methane 2 Modified – 6 0.42 95% Faisal and
emission from POME based on the methodology of Revised 1996 anaerobic Unno, 2001
baffled
Intergovernmental Panel on Climate Change (IPCC) Guidelines for
bioreactor
National Greenhouse Gas Inventories Reference Manual and Workbook 3 Up-flow Mesophilic 3 0.346 97% Najafpour
(Tong and Bakar Jaafar, 2004). The input parameters were listed according anaerobic 38 °C et al., 2006
to the Malaysian palm oil industry in 2009. From the table, total sludge-fixed
film (UASFF)
biomethane emission from POME was estimated at 0.57 million tonnes
4 Single stage Mesophilic 17 0.15a 97% Yacob et al.,
in year 2009. The emission of biomethane in CO2-equivalent corresponds 37–42 °C 2006b
to 11.99 million tonnes, which is equivalent to approximately 6.41% of the 5 Continuous Mesophilic 7 0.30 a
71% Choorit and
total carbon emission in Malaysia in the year of 2006 (187 million tonnes) stirred tank 37 °C Wisarnwan,
(Bernama, 2009). reactor (CSTR) 2007
6 Continuous Thermophilic 5 0.27a 70% Choorit and
Based on previous studies on POME treatment using anaerobic
stirred tank 55 °C Wisarnwan,
digestion system, the end products were mainly consist of biomethane reactor (CSTR) 2007
(CH4) and carbon dioxide (CO2) in 65:35 ratios and approximately 28 m3 a
Self-estimation.
of gases were emitted from 1 tonne of POME (Yacob et al., 2006a).
Nevertheless, biomethane emission may differ depending on treatment
practices as shown in Table 3. From the table, methane emission rate lies temperature and pH play a significant role in determining the
between 0.15 and 0.42 L/g COD removed with COD removal efficiency of performance and fate of the microbial community in anaerobic
70–97%. For a comparison with different feedstocks, mean value of these digesters (Weiland, 2010). Fig. 3 shows the main pathways of an
data were taken as a basis of calculation to standardize the unit reported in anaerobic digestion and the descriptions for each stage are given in
literature (m3 of methane/kg of volatile solids). Fig. 2 shows a comparison the following section. Table 4 displays the overall biochemistry
chart of biomethane emission from different feedstock. From the figure, reaction during the process.
biomethane emission rate from POME was relatively higher as compared
to biomethane emission rate from manure, biomass and municipal solid 4.1.1.1. Hydrolysis. The first step of an anaerobic digestion process is
waste. Thus, there is indeed a huge potential in utilizing the POME to hydrolysis in which organic polymers (carbohydrates, proteins and
generate high commercial return and to produce renewable energy in a lipids) are hydrolyzed to their respective organic monomers. For
more sustainable way. The following section reviews some of the existing example, carbohydrates are converted to sugar or alcohols, proteins to
and recent technologies in anaerobic digestion of POME to produce amino acids and lipids to fatty acids. This is carried out by several
methane. hydrolytic enzymes such as cellulases, cellobiase, xylanase, amylase,
lipase and protease secreted by hydrolytic microbes (Weiland, 2010). The
4.1.1. Biochemistry of anaerobic digestion organic monomers will then be utilized either as substrates by
The metabolic reactions that occur during anaerobic digestion of fermentative organisms (amino acids and sugars) or by anaerobic
substrates involve four important stages: hydrolysis, acidogenesis, oxidizers (fatty acids) (Demirel and Scherer, 2008).
acetogenesis and methanogenesis (Demirel and Scherer, 2008). The
operational efficiency of an anaerobic digestion system primarily
4.1.1.2. Acidogenesis. The second step is acidogenesis (also referred to
depends on the structure of microbial community present in the
as fermentation), in which the hydrolyze products are degraded
system (Weiland, 2010). In addition, environmental factors such as
further to simpler organic products such as acetate, hydrogen (H2)
and carbon dioxide (CO2). These final products of fermentation will
Table 2
Total methane emission from POME in year 2009.
eventually become the precursors of biomethane formation.

Value
4.1.1.3. Acetogenesis. During acidogenesis process, not only acetate, H2
Input parameters and CO2 are produced, but complex intermediary products such as
A Total industrial output (million tonnes/year), crude palm oil 17.56
B Degradable organic component (kg COD/m3 POME)a 51
propionate, butyrate, lactate and ethanol will be produced simulta-
C Wastewater produced (m3 POME/tonnes CPO)b 3 neously. Such intermediary products will be converted to simpler
D Total organic in wastewater (million tonnes COD/year)c 2.69 organic acid, CO2 and H2 by acetogenic bacteria.
E Efficiency of COD removal by anaerobic digestion 0.85

Estimation of maximum methane emission from POME
F Maximum methane producing capacity (kg CH4/kg COD)
G Methane emission from POME (million tonnes CH4/year)d
H Methane emission from POME (million m3 CH4/year at 273 K)
I GHG emission, CO2-equivalent (million tonnes/year)e
a
Assuming COD in POME input to anaerobic digestion at 50,000 mg/L.
b
Assuming 0.6 m3 POME/tonne FFB; 0.2 tonne CPO/tonne FFB.
c
D = A × B × C.
d
G = D × E × F.
e
Global warming potential of methane =21.
4.1.1.4. Methanogenesis. The final step of anaerobic digestion is
0.25 methanogenesis in which methane is produced by two groups of bacteria
0.57 (methanogens), namely acetotrophic methanogens and hydrogeno-
799.61
trophic methanogens. Acetotrophic methanogens convert acetate to
11.99
biomethane (CH4) and CO2 whereas hydrogenotrophic methanogens use
H2 as electron donor and CO2 as electron acceptor to produce biomethane
(Demirel and Scherer, 2008). In addition, many H2-using methenogens
can also use formate as an electron donor for the reduction of CO2 to
biomethene (Demirel and Scherer, 2008). These bacteria are highly
128 M.K. Lam, K.T. Lee / Biotechnology Advances 29 (2011) 124–141

Fig. 2. Methane emission rate from different feedstock (Amon et al., 2007; Chynoweth et al., 1993; Forster-Carneiro et al., 2007; Kabouris et al., 2009; Møller et al., 2004).

sensitive to oxygen; oxygen is a deadly poison that kills all methanogens
even at low concentration.
4.1.2. Conventional ponding (lagoon) system
Ponding system is the most common treatment system used by
Malaysian palm oil mill to treat POME. Surveys conducted by MPOB have
shown that more than 85% of the palm oil mills are using ponding system
due to its low capital cost (Tong and Bakar Jaafar, 2004). Ponding system
consists of a series of anaerobic, facultative, and algae (aerobic) ponds
(Yacob et al., 2009). Facultative and aerobic ponds are necessary to further
reduce the organic content in the wastewater before it is discharged to
rivers (Poh and Chong, 2009). This system requires less energy to operate
due to limited mechanical mixing, operational control and monitoring
(Yacob et al., 2009). However, ponding system requires a large land area to
accommodate a series of ponds in order to achieve the desired
characteristic of discharge.
In the ponding system approach, the depth of pond plays a significant
role in determining the type of biological process. For anaerobic ponds, the
optimum depth ranges from 5 to 7 m (hydraulic retention time (HRT) of
30–45 days) whereas for facultative ponds are 1–1.5 m depth (15–
20 days HRT) (Tong and Bakar Jaafar, 2004; Yacob et al., 2009). A
shallower depth of approximately 0.5–1 m is required for aerobic ponds
with HRT of 24 days (Yacob et al., 2009). Previous studies have shown that
anaerobic ponding system has a higher methane emission rate of

Fig. 3. Anaerobic digestion of organic matter to methane.

 M.K. Lam, K.T. Lee / Biotechnology Advances 29 (2011) 124–141
Table 4
Anaerobic digestion reactions (Denac et al., 1988).
Acidogenesis of glucose
Glucose : C6 H12 O6 →CH3 ðCH2 Þ2 COOH + 2CO2 + 2H2
Glucose : C6 H12 O6 →2CH3 COOH + 2H2 O
Glucose : C6 H12 O6 →2CH3 COOH + 4H2 + 2CO2
Acetogenesis
Butyric acid : CH3 ðCH2 Þ2 COOH + 2H2 O→CH3 COOH + CO2 + 3H2
Propionic acid : CH3 CH2 COOH + 2H2 O→CH3 COOH + CO2 + 3H2
approximately 1043.1 kg/day compared to only 518.9 kg/day from open
digesting tank (Yacob et al., 2006a, 2005). This was attributed to a higher
biomethane composition in the biogas mixture at 54.4%. Nevertheless, the
disadvantages of using ponding system are formation of scum on the
POME surface and solids tend to build-up at the bottom of the pond.
Furthermore, the sludge and scum will clump together inside the pond
and thus lowering the treatment efficiency. Therefore, the system requires
regular desludging process by either using submersible pumps or
excavators to maintain desired efficiency.
4.1.3. Open digesting tanks
Open digesting tanks are used to treat POME when limited land area is
available for ponding system (Poh and Chong, 2009). Normally, an open
digester tank is constructed of mild steel with various volumetric
capacities ranging from 600 to 3600 m3 (Yacob et al., 2009). In addition,
open digesting tank requires shorter HRT ranging from 20 to 25 days
compared to ponding system, 30–45 days. However, treated POME from
the digester will still go through the facultative ponds and followed by
aerobic ponds to further degrade its organic content. Similar to the
ponding system, no mechanical mixing equipment is installed in the
digesters and biomethane is emitted directly to atmosphere (Tong and
Bakar Jaafar, 2004; Yacob et al., 2009). One of the advantages of using
open digester tanks is easier removal of solids build-up at the bottom on a
regular basis, and thus maintaining the desired treatment efficiency
(Tong and Bakar Jaafar, 2004). The dewatered sludge can then be used as
fertilizer. Nevertheless, the major drawback of using open digester tanks
is corrosion of the steel structures due to long exposure to hydrogen
Fig. 4. CSTR anaerobic digester for treatment of POME and methane recovery.
129
sulfide. Incidents such as bursting and collapsed digesters have been
commonly reported (Yacob et al., 2009).
4.1.4. Closed digesting tanks
Closed anaerobic digesting tanks have been developed and
promoted in the last few years under Clean Development Mechanism
(CDM). Under this mechanism, biogas is captured and directly used
for flaring, boiler fuel or power generation (Tong and Bakar Jaafar,
2004). Open pond and open digesting tanks treatment system are
getting less attractive because valuable gas (biomethane) is released
to the atmosphere and those system cannot be certified for Carbon
Emission Reduction (CER) trading (Sulaiman et al., 2009). In fact, a
closed digesting tank has a similar design with an open digesting tank,
except a fixed or floating cover is included and equipped with other
facilities such as gas collector, safety valves and monitoring facilities
(Yacob et al., 2009). Apart from that, a case study on methane
emission rate from a large scale closed digesting tank was conducted
and the emission rate was found to be 5019 kg/day with methane
composition of 62.5%, which is much more higher compared to the
methane emission rate from open pond (1043.1 kg/day) and open
digesting tank (518.9 kg/day) (Tong and Bakar Jaafar, 2004).
Fig. 4 shows a typical closed digesting tank designed by Novaviro
Technology Sdn. Bhd (Tong and Bakar Jaafar, 2004). The system
consist of three continuous stirred tank reactors, in which two
reactors installed with floating roof and another reactor with fixed
cover. The total capacity of the three reactors is 7500 m3 with 18 days
of HRT. Apart from that, the system is equipped with dual-function
mixing mechanism: (1) pump-aided circulation and (2) gas lifting
mixing (Hassan et al., 2009). This anaerobic digesting system has been
perfectly operated continuously for over 20 years. The high efficiency
of the anaerobic digestion design allows maximum recovery of
biomethane possible from the optimally controlled operation of POME
anaerobic digestion. In addition, the total biomethane captured and
utilized as boiler fuel has been estimated to be 1407 tonnes/year,
which is equivalent to 29,547 tonnes/year of CO2-equivalent
reduction.
130 M.K. Lam, K.T. Lee / Biotechnology Advances 29 (2011) 124–141
Fig. 5. Methane captured and utilization in palm oil mill using floating plastic cover in open ponds system.
4.1.5. Other related technologies and future direction
Since conventional anaerobic digestion system requires large
reactor volume (open and closed digesting tanks) and long duration
of HRT, new technologies and reactor design have been proposed to
overcome these limitations. Technologies currently undergoing
intensive research and development include fluidized bed reactor
(Idris et al., 2003), up-flow anaerobic sludge blanket (UASB) reactor
(Borja et al., 1996; Chaisri et al., 2007), up-flow anaerobic sludge
fixed-film (UASFF) reactor (Zinatizadeh et al., 2006a,b, 2007a,b) and
membrane technology (Ahmad et al., 2006a,b, 2009; Wu et al., 2007).
Nevertheless, industrial full scale application of these new reactors
has not been materialized since capital and operating cost are
relatively higher than the conventional anaerobic digestion system.
Furthermore, there is no environment legislation on biomethane
emission from POME or effective biomass policy implemented in
Malaysia to encourage palm oil investors to include renewable energy
as part of their business plan.
On the other hand, a direct and cost effective way to recover
biomethane from POME is to replace the existing ponding/lagoon
system with a closed digester system. This can be simply done by
installing floating plastic membranes on the open ponds. Conse-
quently, biogas will be captured and retained within the floating
plastic covers. Normally, the floating plastic covers are made of linear
low density polyethylene liners that are resistant to bad weather,
biological degradation, UV radiation and can easily evacuate accumu-
lated rainwater. The captured biogas will be removed, cleaned and
combusted as a boiler fuel or to generate electricity. In addition, the
closed anaerobic ponds can be incorporated with biogas recycling
system by pumping back the gas to the bottom of the ponds and thus,
creating a mixing effect in the pond. This will enhances contact
between the POME with microorganisms (particularly methanogens)
resulting to a better organic materials degradation and higher
emission rate of biogas. Effluent from the ponds will be discharged
to facultative and aerobic ponds for further organic matters
degradation. Desludging process for anaerobic ponds will maintain
as normally done by pumping into drying beds and used as organic
fertilizer in agricultural sector. Fig. 5 shows a simplified flow diagram
of biogas utilization in palm oil mill.
Apart from that, biogas refining and storage system can be
installed at palm oil mill for long term investment. Biogas such as
biomethane can be upgraded as transportation fuel and thus enhances
its economic value in renewable energy market. Furthermore,
implementation of such strategy will further strengthen the sustain-
ability of palm oil mill. During biogas refining step, unwanted gaseous
components are removed depending on the requirements and type of
applications as shown in Table 5 (Tong and Bakar Jaafar, 2005). Biogas
can be upgraded to a quality comparable to compressed natural gas
(CNG) in order to be used in vehicles which have been modified to use
CNG. However, the biogas must be able to meet the following basic
requirement with composition of methane content (vol.): N97%; CO2
content (vol.): b3%; H2S: b10 ppmv; and water content: b32 mg/
N m3 (Tong and Bakar Jaafar, 2005). Common technologies available
for biogas upgrading include: water scrubbing, pressure swing
adsorption (PSA) using molecular sieves and membrane separation.
Fig. 6 shows a simple schematic diagram of a gas refinery plant using
water scrubber technology (Tong and Bakar Jaafar, 2005). Crude
biogas is initially pressurized to 8–12 bar and fed to the bottom of the
water scrubber absorption column where CO2 and H2S are absorbed in
water (Tong and Bakar Jaafar, 2005). Water is fed on the top of the
column and the process is operating counter-currently. The exit gas
from the column is dried in an adsorption dryer operating with
zeolites as adsorbing agent. Water is recycled in a stripper or
desorption column. Finally, the refined biogas is compressed to
250 bar and stored in a refueling station ready for use in vehicles
(Tong and Bakar Jaafar, 2005).
Table 5
Requirements to remove gaseous component from biogas.
Application H2S CO2 H2O
Gas heater (boiler) b 1000 ppm No No
Kitchen stove Yes No No
Stationary engine b 1000 ppm No No
Vehicle fuel Yes Yes Yes
Natural gas grid Yes Yes Yes
 M.K. Lam, K.T. Lee / Biotechnology Advances 29 (2011) 124–141
Fig. 6. Biogas refinery flow diagram using water scrubber technology.
4.2. Biohydrogen production
Biohydrogen production from renewable sources such as cellulose-
rich biomass, starch-rich wastes and wastewater has entered a new
dimension in this era as overexploitation of fossil fuels has left mankind
with not much option both from energy and environmental perspective
(Ntaikou et al., 2010). Although hydrogen is not readily available in its
molecular form, it does exist abundantly in nature compared to fossil fuel
(Ntaikou et al., 2010). In addition, hydrogen is a totally clean fuel as
combustion of hydrogen produced only water as by-product (Guo et al.,
2010). As shown in Table 6, hydrogen carries higher energy value per unit
of weight compared to other fuels. In other words, hydrogen can supply
more energy for combustion per unit quantity. These characteristics make
hydrogen an appealing candidate for the future energy system.
Hydrogen can be produced in numerous ways, such as steam methane
reforming (SMR), electrolysis, biomass gasification, biomass pyrolysis,
photofermentation and dark fermentation of waste (Balat, 2009b; Chong
et al., 2009d; Guo et al., 2010). Among these methods, SMR is the only
commercialize method available for hydrogen production due to its
economical efficiency (Jin et al., 2008). Currently, about 48% of the world's
hydrogen production is using this method (Jin et al., 2008). SMR consists
of two steps in which the first step involves methane reforming at
temperature of 500–900 °C and pressure of 30 atm in the presence of
catalyst (Ni) (Balat, 2009b). For the second step, carbon monoxide (CO)
reacts with steam to produce hydrogen and CO2 through a water gas shift
reaction (WGSR). However, SRM reaction is an endothermic process
(Balat, 2009b; Fan et al., 2009). Therefore, external energy must be
supplied to meet the endothermicity of the reaction, which generally
requires combustion of a fuel (Fan et al., 2009). Furthermore, methane
which is mainly derived from fossil fuel has led to unsustainable hydrogen
Table 6
Higher heating value for various fuels.
Fuel Higher heating value (MJ/kg)
Hydrogen 142
Methane 56
Natural gas 54
Gasoline 47
Diesel 45
Ethanol 30
Methanol 23
Coal (anthracite) 27
Wood 15
131
production. The following equations depict the SMR reaction (Balat,
2009b).
Step 1: CH4 + H2O ⇔ CO + 3H2 ΔH = + 206 kJ/mol.
Step 2: CO + H2O ⇔ CO2 + H2 ΔH = − 102 kJ/mol.
Currently, research on hydrogen production has shifted to using
microbial technology via photo fermentation and dark fermentation. For
photo fermentation, microorganisms (Rhodobacter sphaeroides and
Rhodopseudomonas palustris) use carbohydrates or organic acids as
substrates to produce hydrogen and CO2 (Chong et al., 2009d). However,
photo fermentation process suffers various limitations such as low light
conversion efficiency, requirement of high energy demand by nitrogenase
and expensive photo-bioreactors (Hallenbeck and Ghosh, 2009). There-
fore, dark fermentation with acidogenic bacteria (Enterobacter, Bacillus
and Clostridium) is favorable from economic point of view since no direct
sunlight is required, variety of waste streams/energy crops can be used as
subtract and simpler reactor design. Nonetheless, specific control and
monitoring on important parameters such as pH, hydrogen partial
pressure and temperature are required because these variables will
significantly affect the emission rate of biohydrogen (Guo et al., 2010).
Table 7 shows the biological pathways of biohydrogen production
through photo and dark fermentation (Manish and Banerjee, 2008; Su
et al., 2009).
The potential of using raw POME as the main substrate to produce
biohydrogen has been revealed recently (Chong et al., 2009a; Chong
et al., 2009b; Chong et al., 2009c; Ismail et al., 2010; Jamil et al., 2009;
Prasertsan et al., 2009; Yusoff et al., 2009). Naturally, POME contains
hemicelluloses and lignocelluloses material (complex carbohydrate
polymers) which resulted to high COD value (15,000–100,000 mg/L)
(Chong et al., 2009b). Due to this reason, POME can become a suitable
substrate for biohydrogen production and act as a wastewater
treatment process simultaneously. Table 8 shows the yield of biohydro-
gen from POME which lies between 1.1 and 6.9 L H2/L POME depending
on the types of microorganisms and fermentation method. It is obvious
Table 7
Biological process of biohydrogen production.
Photo fermentation Dark fermentation
Glucose as substrate Glucose as substrate; acetic acid as end-product
C6H12O6 +6H2O→12H2 +6CO2 C6H12O6 +2H2O→2CH3COOH+4H2 +2CO2
Acetic acid as substrate Glucose as substrate; butyric acid as end-product
CH3COOH+2H2O→4H2 +2CO2 C6H12O6 →CH3CH2CH2COOH+2H2 +2CO2
132 M.K. Lam, K.T. Lee / Biotechnology Advances 29 (2011) 124–141

Table 8 To date, production of biohydrogen from POME in commercial scale

Yield of biohydrogen production from POME using dark and photo-fermentation is not ready based on current production technology. Apart from safety
technique.
and storage problems of biohydrogen, problems associated with long
No. Microorganism pH Temperature Biohydrogen Reference retention time of dark fermentation and reactor design have been
(°C) yield (L H2/L identified as the main constrains to up-scale biohydrogen production.
POME)
Moreover, most of the researches on biohydrogen production from
Dark fermentation POME only focused on optimization work in lab-scale resulting to
1 Mixed culture 5.5 60 4.6a Atif et al.,
limited information and knowledge gap in commercial scale production.
2005
2 Mixed culture 5.5 60 2.3 Atif et al., A study was conducted previously in a pilot plant using 50 L of CSTR
2005 reactor to produce biohydrogen from POME (Yusoff et al., 2009). The
3 Thermoanaerobacterium 5.5 60 6.3a O-Thong result significantly indicated that lab-scale biohydrogen production
et al., 2007 efficiency did not correlate well when up-scaled to pilot plant. The
4 Thermoanaerobacterium 5.5 60 6.5 O-Thong
biohydrogen emission rate obtained in lab scale and pilot plant was
et al., 2008
5 Clostridium butyricum 5.7 36 6.9a Chong 74 mL/h/L-POME (70% biohydrogen content) and 33 mL/h/L-POME
et al., 2009b (25% biohydrogen content), respectively. These two distinguish
6 Clostridium butyricum 5.5 37 3.4 Chong et al., observations were mainly due to difficulty in pH and temperature
2009c
control during pilot plant operation. Therefore, further investigation and
7 Mixed culture 5.5 23–25 1.2 Yusoff et al.,
2009 research works should be done with special focus on up-scaling
Photofermentation processes, selection of suitable microorganisms, optimization of process
8 Rhodopseudomonas 6.0 – 1.1 Jamil et al., conditions and effective parameters control.
palustris 2009 Another potential approach to conceptualize commercial scale of
a
Self-estimation. biohydrogen production is through integration of dark and photo
fermentation. The objectives of this integration are to overcome the
limitations of individual fermentation methods and for further
enhancement of energy recovery by maximizing the yield of
that dark fermentation is more favorable than photofermentation with biohydrogen (Brentner et al., 2010). For the first phase of the
higher biohydrogen yield of 4.5 L H2/L POME (mean value). This is integrated system, dark fermentation is carried out to break down
because limited light penetrations caused by the sludge particles reduce the complex carbohydrate polymers from POME to organic acids such
the conversion efficiency of photo fermentation microorganisms. Base as acetic and butyric acid. Consequently, the organic acids will be
on 43,800 million L of POME generated in 2009, it was estimated that subjected to photo fermentation to further breakdown the organic
197,100 million L of biohydrogen can be produced. If this amount of acids and to produce biohydrogen. Theoretically, a maximum yield of
biohydrogen is successfully captured and utilized as energy, the energy 12 mol biohydrogen per mol of glucose could be achieved through this
attained is equivalent to 2.51 PJ in which contributes to 6.5% of the integrated dark-photo fermentation (Das, 2009), while 8 mol of
projected renewable energy demand (375 MW) in Malaysia in year biohydrogen per mol of glucose is considered sufficient for economic
2010 (Oh et al., 2010) (assuming 1 PJ can be converted into 46 MW of applications (Chen et al., 2010). On the other hand, apart from photo
energy power with 21% conversion efficiency (Tock et al., 2010)). Apart fermentation, dark fermentation also can be integrated with metha-
from that, the potential of biohydrgen derived from POME is further nogenic process whereby biomethane is produced after dark fermen-
strengthened by referring to Fig. 7 that shows the yield of biohydrogen tation. Such strategy offers several advantages than integration with
from POME is the highest in comparison with other wastes and photo fermentation in terms of waste stabilization and organic
wastewaters. Nevertheless, it should be noted that the figure presents a degradation (Ntaikou et al., 2010). A combined biohydrogen and
rough indication, regardless to their fermentation method, type of biomethane generation process has been proposed for organic solid
microorganisms used and reactor design. wastes (Ueno et al., 2007), food wastes (Han and Shin, 2004), cheese

Fig. 7. Yield of biohydrogen by using different feedstock (Fang et al., 2006; Kim and Lee, 2010; Ntaikou et al., 2009; Pan et al., 2008; Van Ginkel et al., 2005; Venetsaneas et al., 2009;
Xing et al., 2010; Zhang et al., 2003; Zhu et al., 2009).
 M.K. Lam, K.T. Lee / Biotechnology Advances 29 (2011) 124–141
whey (Antonopoulou et al., 2008), olive mill wastewater (Koutrouli et
al., 2009) and wastewater sludge (Ting and Lee, 2007).Up to now,
integrated system for POME treatment, neither dark-photo fermenta-
tion nor dark fermentation-methanogenic process has not been
investigated. Thus, there is an urgent need to fill up the gap knowledge
since this integrated system is the key step to commercial viability and
sustainable energy production from POME. Fig. 8 shows a conceptual
flow diagram of an integrated system for biohydrogen production
coupled with methanogenic process.
4.3. POME cultured microalgae: a novel approach to produce biofuels
Sustainable energy management in palm oil mill has entered a new
dynamic era with the possibility of culturing microalgae using POME.
In the last few years, microalgae have been highlighted as the most
feasible alternative feedstock for biofuels production due to its fast
growth rate and ability to synthesis large quantity of neutral lipids/oil
(Khan et al., 2009). In addition, microalgae can provide different types
of biofuels, such as biodiesel derived from microalgae lipid; bioethanol
produced from fermentation of microalgae biomass; biomethane
produced by anaerobic digestion of microalgae biomass; and photo-
biologically produced biohydrogen (Chisti, 2007). The following
section reviews the current status of biofuels production from
microalgae and the potential of using POME as nutrients source.
4.3.1. Microalgae
Microalgae are recognized as one of the oldest living microorganisms
on Earth (Song et al., 2008). They are single cell organisms,
representative of both bacteria and eukaryotes (Williams and Laurens,
2010). A significant characteristic that distinguish between bacteria and
eukaryotes is that the former lack of discrete internal, sub-cellular
structures, organelles (chloroplasts, mitochondria and nuclei) (Williams
and Laurens, 2010). Eukaryotes, which comprise of many different types
of common microalgae, do have organelles that control the functions of
the cell, allowing it to survive and reproduce (Brennan and Owende,
2010). To date, microalgae species are divided into four categories:
diatoms (Bacillariophyceae), green algae (Chlorophyceae), blue-green
algae (Cyanophyceae) and golden algae (Chrysophyceae), depending on
their pigmentation, life cycle and basic cellular structure (Khan et al.,
2009). Carbohydrates, proteins, nucleic acids and lipids (typically
Fig. 8. Integrated system of dark fermentation coupled with anaerobic digestion for biohydrogen and biomethane production.
133
phospholipids and glycolipids) are the major constituents of microalgae
(Williams and Laurens, 2010).
One of the reasons that microalgae appear to be an attractive
renewable energy source is due to its rapid growth rate; 100 times
faster than land-based plant and they can double their biomass in less
than 1 day (Tredici, 2010). Furthermore, microalgae are able to divide
once every 3–4 h, but mostly divide every 1–2 days under favorable
growing conditions (Williams and Laurens, 2010). This is mainly due
to their simple cellular structure and large surface to volume ratio that
give them the ability to uptake large amount of nutrients from water
sources and thus, promoting their growth rate (Khan et al., 2009).
Apart from that, microalgae can be cultured either autotrophic or
heterotrophic. Autotrophic microalgae such as Botryococcus braunii
and Dunaliella salina require sunlight, CO2 and nutrients as a basic
requirement for growing whereas heterotrophic microalgae such as
Chlorella protothecoides require organic carbons sources (sugar and
organic acids), CO2, nutrients but independent of sunlight (Liang et al.,
2009). Note that C. protothecoides can be grown autotrophically or
heterotrophically under different culture conditions. However, het-
erotrophically growth of C. protothecoides is favorable due to higher
biomass production and accumulation of higher lipid content in cells
(Miao and Wu, 2006).
The potential of microalgae cultivation for biofuels production can
be clearly seen in Table 9. From the table, culturing microalgae (either
high or low lipid content) required the least land area in comparison
with other oil crops such as soybean, sunflower, rapeseed and oil palm
in order to meet European Union (EU) biofuels target in year 2010. In
addition, using edible oils to produce biofuels is recognized as an
unsustainable way due to the arising public concern on “food versus
fuel” issue. Therefore, to find alternative feedstock for biofuels
production, focus has recently being directed to using non-edible oil
specifically, Jatropha curcas L., a type of non-food crop which is easily
grown on wasteland (Lam et al., 2009b). However, regular irrigation,
heavy fertilization and good management practices are mandatory
steps to ensure high seeds yield production from jatropha. Up to now,
the highest jatropha seeds production rate was reported to be
12 tonnes/ha/year in which equivalent to 4.13 tonnes of oil/ha/year
(assuming oil content of 34.4%) (Lam et al., 2009b). This value is still far
lower than microalgae with an annual lipid production rate of 54–
126 tonnes/ha/year. Base on this evidence, microalgae seems to be a
134 M.K. Lam, K.T. Lee / Biotechnology Advances 29 (2011) 124–141

Table 9 Table 11
Comparison of oil yield for various oil bearing plants and microalgae (Chisti, 2007; Lam Fatty acid composition for microalgae and other oil crops (Balat and Balat, 2010;
et al., 2009b; Lam et al., 2009c). Karmakar et al., 2010; Ma and Hanna, 1999; Šimáček et al., 2009; Yan et al., 2010).

Oil crop Average oil yield Area to meet EU biofuel % of current Fatty acid Number of Oil composition (%)
(tonne/ha/year) demand in 2010 Malaysian's carbon atoms:
Microalgae Soybean Rapeseed Palm Jatropha
(million ha)a agricultural land areab number of
double bonds
Soybean 0.4 25.0 379
Sunflower 0.46 21.7 329 Lauric C12:0 0 0.1 0 0.1 0
Rapeseed 0.68 14.7 223 Myristic C14:0 2.7 0.1 0.1 1 0
Oil palm 3.62 2.8 42 Palmitic C16:0 20.9 13.1 4.8 42.8 14.2
Jatrophac 0.14 71.4 1082 Palmitoleic C16:1 10.6 0 0.2 0.23 0.7
Jatrophad 4.13 2.4 37 Stearic C18:0 6.9 3.7 1.9 4.5 7
Microalgaee 126 0.1 1 Oleic C18:1 33.3 22.8 61.9 40.5 44.7
Microalgaef 54 0.2 3 Linoleic C18:2 18.5 53.7 19.8 10.1 32.8
a Linolenic C18:3 1.2 6.6 9.2 0.2 0.2
EU biofuel target in year 2010 is equivalent to 10 million tones.
b Others – 6.9 0 2.2 0 0
Total agricultural land area in Malaysia is equivalent to 6.6 million ha.
c
Jatropha are planted without irrigation and fertilization.
d
Jatropha are planted with irrigation and heavy fertilization.
e
Microalgae synthesized high lipid content (70% lipid based on biomass weight). production (Clarens et al., 2010). High requirement of nitrogen and
f
Microalgae synthesized low lipid content (30% lipid based on biomass weight). other related chemical fertilizers to culture microalgae on a large scale
have driven the process towards non-environmental friendly. On the
superior feedstock for biofuels production. An added advantage to other hand, culturing microalgae can actually play an important role
microalgae biofuels is it does not compete with food production and as a self-purification process of natural wastewaters (Soeder, 1980).
thus, up hold an important key for a more sustainable energy Conventional treatment of municipal wastewater that involves
development in the future. primary and secondary biological treatment only managed to remove
Table 10 shows the lipid content of various microalgae strains. a fraction of nitrogen and phosphorus contained in the wastewater
From the table, lipid content for freshwater and marine microalgae (Órpez et al., 2009). Therefore, culturing of microalgae in wastewater
lies between 20 and 75% (weight of dry biomass). In addition, oil offers an inexpensive alternative to conventional forms of tertiary
productivity (mass of oil produced per unit volume of the microalgae wastewater treatments and at the same time to utilize the nitrogen
broth per day) is dependent on the algae growth rate and the oil and phosphorus compound in wastewater to generate microalgae
content of the biomass (Chisti, 2007). As mentioned previously, biomass for biofuels production. In term of environmental and
microalgae lipid contains high concentration of phospholipid and economic point of view, such win–win strategy will be the most
glycolipid. These lipid groups contain nitrogen, phosphorus and sulfur plausible option to commercialize microalgae cultivation process.
that may be problematic with regards to engine performance. Utilizing POME as nutrients source to culture microalgae is not a
Nevertheless, nitrogen, phosphorus and sulfur containing compound new scenario, especially in Malaysia. Most palm oil millers favor the
can be minimized through water soluble fraction and followed by culture of microalgae as a tertiary treatment before POME is
transesterification reaction. Therefore, one would expect that micro- discharged due to practically low cost and high efficiency. Therefore,
algae biodiesel will not deteriorate diesel engine combustion most of the nutrients such as nitrate and ortho-phosphate that are not
efficiency. Table 11 shows the lipid composition of microalgae in removed during anaerobic digestion will be further treated in a
comparison with other oil bearing crops. microalgae pond. Consequently, the cultured microalgae will be used
as a diet supplement for live feed culture. Table 12 shows the
characteristics of POME after secondary treatment. From the table, the
4.3.2. POME as nutrients source to culture microalgae
total nitrogen contents are still high and do not meet the discharge
Realizing the huge potential of culturing microalgae towards
standard for wastewater which is 200 mg/L. Meanwhile, nitrogen
sustainable energy production, researchers in this field have identified
source (usually appears in nitrate form) plays an important role in
a few key points in up-scaling the whole process. One of the obstacles
promoting microalgae growth. In order to grow microalgae effectively,
in up-scaling microalgae culture to industrial scale is the source of
the basic nitrate concentration required is in the range of 200–400 mg/
nutrients in the culture medium. A life cycle assessment on
L (Li et al., 2008a). Others minerals such as Fe, Zn, P, Mg, Ca and K that
microalgae cultivation has underlined that 50% of energy use and
are required for microalgae growth are also present in POME. (Habib et
green house gases emissions are associated with fertilizer (nutrients)
al., 1998).Thus, POME emerged to be an alternative option as a
chemical remediation to grow microalgae for biomass production and
Table 10 simultaneously act as part of wastewater treatment process.
Lipid content for various freshwater and marine microalgae (Chisti, 2007; Rodolfi et al.,
2009).

Habitat Microalgae Lipid content (% dry wt)

Table 12
Freshwater Botryococcus braunii 25–75 Characteristics of POME after stirred digester tank and lagoon pond.
Chlorella sp. 28–32
Neochloris oleoabundans 35–54 Parameter Final dischargea
Phaeodactylum tricornutum 20–30
Range Mean
Scenedemus sp. 20–21
Marine Crypthecodinium cohnii 20 pH 7.0–7.4 7.3
Cylindrotheca sp. 16–37 Oil and grease 10–430 130
Dunaliella primolecta 23 BOD 110–1800 610
Isochrysis sp. 25–33 COD 340–19,680 4820
Monallanthus salina N 20 Total solid 6090–18,400 10,360
Nannochloris sp. 20–35 Suspended solid 760–14,850 4680
Nannochloropsis sp. 31–68 Volatile solid 2680–14,570 5000
Nitzschia sp. 45–47 Total nitrogen 320–1070 520
Schizochytriukm sp. 50–77 Ammoniacal nitrogen 60–300 180
Tetraselmis sueica 15–23 a
All parameters are in mg L− 1 except temperature and pH.
 M.K. Lam, K.T. Lee / Biotechnology Advances 29 (2011) 124–141
Fig. 9. Overview design of POME treatment incorporating with microalgae culture.
4.3.3. Overall system design
A conceptual design of incorporating microalgae farm fed by POME
is shown in Fig. 9. The anaerobic digestion process and biomethane
production was already discussed previously. Therefore, the following
part will describe the microalgae cultivation, microalgae biomass
harvesting and biofuels production process. Apart from that, GHG
mitigation strategy using microalgae in palm oil mills will be
elaborated as well. Through this integrated system, sustainable and
renewable energies production from palm oil mill can be achieved
simultaneously with air pollution control and wastewater treatment.
4.3.4. Open pond and closed photo-bioreactor system
The principle of microalgae cultivation requires sufficient supply of
sunlight, CO2 and nutrients (Brennan and Owende, 2010). Therefore, a
comprehensive design of cultivation system is important to ensure
optimum microalgae growth condition and produce large quantity of
biomass. Currently, phototrophic method (sunlight) is the only
technical and economical feasible way to culture microalgae in large
scale (Borowitzka, 1999). Generally, phototrophic method can be
divided into two types; open pond system and closed photo-
bioreactor system (Chisti, 2007; Greenwell et al., 2010). The following
section details out the basic design of the two systems.
4.3.4.1. Open pond system. The commonly used method to culture
microalgae in large scale is the open pond system. This is because open
pond system offers a relatively lower cost to built and easy operation
(Mata et al., 2010). Among all the open pond system, raceway pond
design is usually applied to ensure good nutrient homogenizing and to
avoid microalgae biomass sedimentation. Fig. 10 shows the conceptual
design of the raceway pond. Generally, the pond consists of closed-
loop recirculation channel (oval shape) in which mixing and
circulation are provided by paddle wheel (Chisti, 2007; Greenwell et
al., 2010). Apart from that, the depth of the pond is usually 0.2 to 0.5 m
to ensure microalgae received adequate exposure to sunlight
(Brennan and Owende, 2010). In addition, raceway pond is built
using concrete, but compacted earth-lined pond with white plastic has
also been used (Brennan and Owende, 2010; Chisti, 2007). Although
open pond system is low in cost, however this system suffers from
several limitations. The main disadvantage of using open system is
being exposed to atmosphere resulting to lost of water due to water
evaporation and susceptible to environment and undesired micro-
135
organisms contamination (Schenk et al., 2008). Over the period of
operation, undesired microorganisms are inevitably introduced and
thus, potentially to dominate in the raceway pond (Schenk et al.,
2008). Furthermore, contamination becomes more serious if POME is
utilized as the nutrients source. This is because POME contained high
value of BOD and COD that may be suitable for other microorganisms
to grow. Due to this phenomenon, microalgae biomass yield will be
reduced. Hence, regular cleaning and maintenance are required to
ensure the system operates at optimum condition.
4.3.4.2. Closed photo-bioreactor system. Closed photo-bioreactor offers
several advantages compared to open pond system, in term of
reducing contamination level, higher microalgae biomass yield with
respect to reactor volume and permit essentially single-species culture
of microalgae for prolonged durations (Chisti, 2007; Greenwell et al.,
2010). Normally, closed photo-bioreactors are designed in tubular,
plate and column shape (Brennan and Owende, 2010) as shown in
Table 13. Apart from that, closed photo-bioreactor is a more flexible
system in which the reactors can be optimized according to the
biological and physiological characteristics of the microalgae strains
being cultivated (Mata et al., 2010). Furthermore, microalgae growth
parameters such as pH, temperature, concentration of CO2 and
nutrients can be controlled. However, the main drawback in convert-
ing this closed photo-bioreactor systems to commercial scale is the
high operating costs compared to open ponds systems (Carvalho et al.,
2006).
Fig. 10. Raceway pond for microalgae cultivation.
136 M.K. Lam, K.T. Lee / Biotechnology Advances 29 (2011) 124–141

Another potential way to up-scale the closed photo-bioreactor

–The columns are usually made of transparent plastic or glass

–Cell sedimentation may occurred if airlift system is not used

system is by using the secondary treated wastewater as culture
medium and nutrients source. For the case of using POME as nutrients

–Optimum column diameter is 0.2 m with 4 m height

–Bioreactor with vertical arranged cylindrical column
source, there is definitely a plausible opportunity to establish a closed

–Usually coupled with a pump or airlift technology

–Compact, easy to operate and relatively low cost

photo-bioreactor systems in palm oil mills. Palm oil mills are usually
energy self-sufficient. Oil palm biomass (fiber and shell) are burnt as

–High mass transfer rate with good mixing

boiler fuel to generate steam and electricity for sterilization process
and power supplement. For some cases, electricity is even supplied to
the nearby estates (Lam et al., 2009b; Subramaniam et al., 2008).

–Small illumination surface area

Therefore, an alternative way to utilize this additional energy

–Lower power consumption

(electricity) is by using it for closed photo-bioreactor system
operation, such as pumps, temperature control systems and artificial
illumination (during night time). It is strongly believe that by
Vertical-column

implementing this strategy, operation cost for closed photo-bioreac-

tor systems can be reduced significantly. Nevertheless, this statement
needs to be further supported through a comprehensive life cycle
assessment study.

4.3.5. Mitigation of CO2 from palm oil mill by microalgae cultivation

CO2 emission in palm oil mills is crucial since fibers and shells are
–Tube diameter is limited (0.1 m) to increase the surface/volume ratio

burned to generate energy. Fibers and shells contained high amount

–Consists of array of straight, coiled or looped transparent tubes

of carbon element, 47.2% and 52.4%, respectively (Mahlia et al., 2001);

(poison to microalgae) in culture medium if tubes are too long

if they are burned, 13.7–161.2 kg of CO2 will be emitted to atmosphere

–Potential cell damage is minimized if airlift system is used
–The tubes are usually made of transparent plastic or glass

to produce 1 tonne of crude palm oil (Subramaniam et al., 2008).

–Required large land space due to long tubes are used

Furthermore, if CO2 is emitted from POME and not collected, total CO2
–Potentially to accumulate high concentration of O2

–Decreasing of CO2 gradient resulted to microalgae

–Usually coupled with a pump or airlift technology

emission from palm oil mill will reach up to 68.2–279.6 kg CO2 per
tonne of crude palm oil. Thus, it was estimated that 1200–
4900 million kg of CO2 was emitted based on palm oil production in
–Mixing is problematic in extended tubes
–Relatively higher biomass productivity

year 2009. Note that this amount of CO2 emission are limited to palm
oil milling process, excluding methane emission from POME and oil
–Large illumination surface area

palm photosynthesis rate (utilizing back the CO2). Therefore, there is

starvation for carbon source

an urgent need to control serious CO2 emission in palm oil mills to

ensure its environmental sustainability. One of the effective ways is
Various design of closed photo-bioreactor system (Chisti, 2007; Sierra et al., 2008; Ugwu et al., 2008; Xu et al., 2009).

through microalgae cultivation.

Microalgae is a fast growing microorganism with an ability to fix
CO2 from atmosphere, flue gases or soluble carbonate while capturing
solar energy with efficiency 10–50 times greater than terrestrial
Tubular

plants (Khan et al., 2009; Li et al., 2008b). The best method for
microalgae to act as carbon sink is to capture CO2 from the
atmosphere for its growth; however this method is limited by low
CO2 concentration in the air (360 ppm) (Brennan and Owende, 2010)
–The flat-plate are usually made of transparent plastic or glass

–Scale-up require many compartments and support materials

which makes it economic infeasible. This is further hindered by the

–The light path (depth) is dependent on microalgae strain;

–Can be positioned vertically or inclined at am optimum

low mass transfer rate of CO2 in water and thus, higher air pumping
–Large illumination surface area resulted to maximum

cost is required. On the other hand, microalgae have a better

adaptability to high concentration of CO2 up to 20% from flue gases
emission (Brennan and Owende, 2010). Nevertheless, some micro-
–Difficulty in controlling culture temperature

algae cannot withstand the acidic condition in culture medium

–Low concentration of dissolved oxygen

resulted from carbonic acid (weak acid) created from CO2 dissolution
–Bioreactor with rectangular shape

in water. Moreover, only a small number of microalgae strains are able

–Usually coupled with gas sparger

to tolerate high levels of SOx and NOx that are present in flue gases
ranged between 1.3 and 10 cm

–Lower power consumption

(Brennan and Owende, 2010). Up to now, Chlorella sp., Scenedesmus

utilization of solar energy

sp. and Botryococcus braunii were determined as potential microalgae

to biomitigate CO2 from flue gas while producing high lipid content
angle facing the sun

for subsequent biodiesel production (Yoo et al., 2010). Furthermore, it

was found that Scenedesmus sp. and B. braunii grew better using flue
gases in comparison with air enhanced with CO2 (Yoo et al., 2010). In
Flat-plate

other words, there is still a huge potential to utilize flue gases from
palm oil mill to culture microalgae coupled with POME as nutrients
source, since Scenedesmus sp. and B. braunii are freshwater microalgae
that can be easily found in tropical countries like Malaysia. Thus this
Design characteristic
Conceptual design

potential should be exploited in the near future through intensive

Disadvantages

research works.
Advantages

4.3.6. Microalgae biomass harvesting method

Table 13

Microalgae biomass harvesting process always posed a challenging

task to engineers typically in commercial scale production. This is
 M.K. Lam, K.T. Lee / Biotechnology Advances 29 (2011) 124–141 137

because microalgae are small size microorganism with diameter extremely fast growth rate compared to terrestrial crops, high lipid
ranging from 3 to 30 μm (Molina Grima et al., 2003) and therefore content and less requirement of land area (Chisti, 2007).
biomass recovering process becomes a very challenging task. Apart
from that, the harvesting process is a solid–liquid phase separation
O O
that requires a lot of energy to recover the biomass efficiently and to
dehydrate the biomass after harvesting process. Generally, microalgae CH2-O-C-R1 CH3O-C-R1 CH2-OH
harvesting process accounted for 20–30% of the total cost of O O (1)
production (Ma and Hanna, 1999) (Gudin and Therpenier, 1986),
CH-O-C-R2 + 3CH3OH CH3O-C-R2 + CH-OH
depending on the harvesting method. Microalgae biomass can be O O
harvested via several conventional methods, such as centrifugation,
flocculation, floatation, filtration and gravity sedimentation. A more CH2-O-C-R3 CH3O-C-R3 CH2-OH
advanced approach is through ultrasonication process. Table 14
summarized the current available microalgae harvesting method. Triglyceride Methanol Methyl Ester Glycerol

After microalgae biomass are harvested from open pond or

4.3.7. Biodiesel production from microalgae
Biodiesel is an alternative diesel fuel that offers several advantages
to environment, such as biodegradable, non-toxicity, better lubricity,
low SOx and CO emission (Jacobson et al., 2008). Normally, biodiesel is
produced through transesterification reaction in which oil reacts with
short chain alcohol (e.g. methanol) in the presence of catalyst (e.g.
sodium hydroxide, NaOH) as shown in Eq. (1) (Sharma and Singh,
2009). For first generation biodiesel production, edible vegetable oils
such as soybean, rapeseed, sunflower and palm oil are used as the
main feedstock. However, with increasing concerns from public
regarding the “food versus fuel” issue, second generation biodiesel
produced by using non-edible oil such as J. curcas L. emerged as a new
alternative for the biodiesel industry. Although jatropha tree can be
planted on non-fertile land but heavy irrigation and fertilization are still
required to ensure high oil yield (Lam et al., 2009b; Openshaw, 2000). In
addition, competition of land area to produce raw oil for biodiesel
production still remains un-resolved. Therefore, for third generation
biodiesel, microalgae is proposed as a potential feedstock due to its
photobioreactor, the biomass is then subjected to dehydration
process. The most economic dehydration method is through sun
drying but this method requires long drying duration, large drying
area and possible contamination from surrounding (Brennan and
Owende, 2010; Prakash et al., 1997). Alternative ways such as spray
drying and freeze drying are only available for small scale usage due to
their relatively high operating cost (Brennan and Owende, 2010).
After the dehydration process, the dried microalgae biomass then
undergoes cell destruction process in order to extract lipid out from
microalgae cell. Various lipids extraction methods have been
reported, such as mechanical press machine, solvent extraction,
supercritical fluid extraction and ultrasound technology as shown in
Table 15. However, only mechanical press machine is economically
feasible in the commercial scale due to its relatively low operating
cost and easy maintenance. On the other hand, solvent extraction
method (hexane, methanol coupled with chloroform) seems to be
another efficient way to extract lipids from microalgae biomass.
Nevertheless, issues such as overall energy efficiency, wastewater

Table 14
Microalgae biomass harvesting methods (Brennan and Owende, 2010; Greenwell et al., 2010; Molina Grima et al., 2003; Schenk et al., 2008).

Harvesting Process description Advantages Disadvantages

method

Centrifugation -Described by Stokes' law — sedimentation of -Rapid and efficient -Energy intensive and high maintenance cost
suspended solids is determined by density
and radius of microalgae cells
-Concentrated microalgae biomass can be obtained
-Centrifugation force — 5000–10,000 g with
95% removal efficiency
Flocculation -To aggregate the microalgae biomass to a larger -Cost effective -The microalgae biomass cannot be used
size and hence ease sedimentation. for some downstream applications such as
-Flocculants used: Ferric chloride (FeCl3), aluminum animal feed supply or for anaerobic digestion
sulfate (Al2(SO4)3, alum) and ferric sulfate (Fe2(SO4)3)
-Normally applied as pre-step before centrifugation,
gravity sedimentation and filtration
Floatation -Trapping microalgae biomass by dispersed micro air bubbles -Applicable to process large volume of -Contamination with the flocs agent and
-The fine bubbles less than 10 μm adhere the biomass biomass may decrease microalgae biomass value
(pre-step is flocculation) making them very buoyant and
causing them rise rapidly to the surface.
Filtration -Filter press and membrane filter (micro and ultrafiltration) -Filter press: effective in recovering -Filter press: Ineffective to recover small size
are operated under pressure or vacuum condition relatively large size microalgae microalgae (e.g. Scenedesmus and Chlorella)
(e.g. Spirulina platensis)
-Micro/ultrafiltration: effective in -Micro/ultrafiltration: High cost due to
recovering large and small size microalgae membrane replacement, membrane
clogging and maintenance
Gravity –Described by Stokes' law — sedimentation of -Low cost as no additional chemicals or -Required relatively longer settling time
sedimentation suspended solids is determined by density and physical treatment -Not effective for small size microalgae
radius of microalgae cells
–Microalgae are left to settle naturally by the
means of gravity
Ultrasonication –Ultrasound wave (20–100 MHz) can compresses -Can be operated continuously without -Safety problem
and stretches molecular spacing of a medium in which inducing shear stress on the microalgae
it passes through and hence creates cavitations effect biomass
–Microalgae cell are destructed immediately and thus
facilitated sedimentation rate
138 M.K. Lam, K.T. Lee / Biotechnology Advances 29 (2011) 124–141
Table 15
Microalgae lipid extraction methods (Couto et al., 2010; Harun et al., 2010).
Extraction methods Process description
Mechanical -Physical pressure is introduced to
press machine microalgae biomass and compress out lipids
-Without chemical additive
-Expeller press (screw-type machine)
is normally used
Solvent -Solvent such as hexane is introduced
extraction to microalgae biomass
-Solvent will destroy microalgae cell wall
and lipids are extracted out
-Normally, lipids have a higher solubility in
organic solvent
-Distillation is required as the final step to recover
the solvent for subsequent extraction cycle
Supercritical -Supercritical fluid such as CO2 and water are
fluid extraction commonly used
-High temperature and pressure are generally required
Ultrasound -High intensity ultrasonic wave is introduced to microalgae
technology -Cavitations create by ultrasound will breaks the microalgae
cell and releasing lipids into solution
treatment problem, solvent purification and storage must be
addressed before this method can be implemented in commercial
scale. In addition, combination of mechanical pressing and solvent
extraction is another possible way to extract more lipids from
microalgae biomass. Supercritical fluid extraction and ultrasound
technology are new approaches proposed by researchers to facilitate
the lipid extraction process (Mata et al., 2010). More research work
and investigation are urgently required to justify the possibility of
these new technologies in term of cost effectiveness and environ-
mental related issues.
The extracted lipids are then subjected to transesterification
reaction in which biodiesel is produced as the main product whereas
glycerol is the by-product. During transesterification, homogeneous
base catalysts such as NaOH and KOH are generally used to facilitate
the reaction towards completion at mild reaction temperature (50–
80 °C) (Lam et al., 2010). In addition, these catalysts are widely used
in commercial scale biodiesel processing plants due to their relatively
cheap price and easily available in the market. However, these
catalysts are sensitive to the free fatty acid (FFA) in raw oil in which
the FFA will react with the catalysts to form soap (Lam et al., 2010).
The soap formed will eventually lower down the yield of biodiesel and
creates difficulty in product separation and finally resulting to large
amount of wastewater generated during biodiesel purification step.
Since microalgae lipids contain high FFA, homogeneous base catalyst
is not a suitable choice unless the lipids are pre-treated with acidic
catalysts (Greenwell et al., 2010). During the pre-treatment step
(esterification reaction), strong acidic catalysts such as sulfuric acid
(H2SO4) or hydrochloric acid (HCl) are used to minimize the FFA
content in the oil before transesterification reaction takes place (Lam
et al., 2010). At this moment, two steps process involved esterification
and transesterification is the most possible way to be implemented in
large-scale microalgae biodiesel production. However, more attention
and proper process design should be focussed on the pre-treatment
step as strong acidic catalysts are categorized as hazardous chemicals
and potentially caused serious corrosion to valves, pipelines and
reactor vessels.
Heterogeneous acid catalysts such as sulfated zirconium oxide (SO2−
4 /
ZrO2) (Furuta et al., 2004; Jacobson et al., 2008; Jitputti et al., 2006),
sulfated titanium oxide (SO2−
4 /TiO2) (de Almeida et al., 2008; Shu et al.,
2009), sulfated tin oxide (SO2−
4 /SnO2) (Furuta et al., 2004; Lam et al.,
2009a), sulfonic ion-exchange resin (Dos Reis et al., 2005; Vicente et al.,
Advantages Disadvantages
-Relatively lower operating cost -Relatively low extraction efficiency
-Easy operation
-Quality of oil cake from pressing is
higher, suitable for downstream process.
(e.g. fertilizer, fermentation to bioethanol)
-High extraction efficiency -Extra energy is required to
-Solvent can be recycled to use recover the solvent
-The use of chemicals present safety
and health issue
-Short extraction time: mass transfer is highly -Energy intensive: high temperature
facilitated due to high mass and thermal and pressure are required
diffusivity coupled with low viscosity -The left behind biomass may not
-Contamination of final product does not occur suitable for downstream process as
most of the carbohydrates and
proteins are destroyed
-Shorten extraction time -High power consumption
-Reduce solvent consumption -Safety and health issue
1998), and sulfonated carbon-based catalyst (Hara, 2009; Lou et al., 2008;
Takagaki et al., 2006) seem to have lot of potential to produce biodiesel
from high FFA oil, typically microalgae lipids. Heterogeneous acid
catalysts offer several advantages compared to homogeneous base
catalysts: (1) esterification and transesterification can be performed in a
single step, (2) insensitive to FFA content in oil, (3) easy separation of
catalyst from reaction products and (4) catalysts can be recycled and
regenerated for the subsequent reaction cycle (Kulkarni and Dalai, 2006).
Apart from that, these catalysts hold an important key to be
incorporated into a continuous flow reactor in which product
separation and purification step can be minimized (Lotero et al.,
2005). Thus, economic feasibility of biodiesel production is improved
and even able to compete with commercial petroleum-based diesel fuel
(de Almeida et al., 2008). Nevertheless, the bottlenecks to commer-
cialize these catalysts are due to its high costs, complicated synthesis
procedures and leaching of catalyst active sites may result to product
contamination. Furthermore, high reaction temperature, high molar
ratio of alcohol to oil and long reaction time are generally required to
drive transesterification reaction towards completion (Lam et al.,
2010). Currently, heterogeneous acid catalysts are still in the research
stage and commercialization of these catalysts in biodiesel production
will be a matter of time. Research focus on heterogeneous acid catalysts
should shift toward implementation of these catalysts in up-scaled
biodiesel plant rather than catalyst synthesis procedures and reaction
optimization in lab scale.
Apparently, biodiesel production from microalgae is more focused
on large-scale cultivation, effective photo-bioreactor design and
increase lipids productivity through optimization work. Nevertheless,
more attention on technologies to convert microalgae lipid to
biodiesel is extensively required to further strengthen the sustain-
ability of microalgae biodiesel. Up to now, only homogeneous base
catalysts are favoured to be used to produce microalgae biodiesel
mainly due to its easy preparation step and fast reaction rate. Other
catalysts such as heterogeneous acid catalysts and enzymatic catalysis
are still limited in the literature. Apart from that, there is still a
knowledge gap on ultrasonication, supercritical fluid and microwave
technologies to enhance microalgae lipid conversion to biodiesel that
can improvement mass transfer rate. In other words, microalgae
biodiesel conversion processes now emerge as a new dimension in
research and development to further enhance the overall process in
term of environmental and economic perspective.
 M.K. Lam, K.T. Lee / Biotechnology Advances 29 (2011) 124–141 139

4.3.8. Bioethanol production from microalgae
Bioethanol is another attractive renewable fuel. It is non-petroleum
based and commonly used as a substitution to gasoline. Although
bioethanol has a lower calorific value than gasoline (22 MJ L− 1 and
33 MJ L− 1), however the higher octane number of bioethanol and the
adjustments to the engines and injection systems resulted to a higher
technical equivalent of bioethanol per liter of gasoline, which is about 1.15
(Costa and de Morais, 2011). Apart from that, bioethanol offers several
advantages compared to gasoline such as broader flammability limits,
higher flame speed and higher heat of vaporization (Balat, 2009a). In term
of environmental perspective, bioethanol has a lower emission of CO2, CO,
lead, sulfur and particulate when combusted in engine (Costa and de
Morais, 2011).
Bioethanol can be derived from: (1) sugary feedstock such as sugar
cane and sugar beet, (2) starchy materials such as corn and wheat and
(3) lignocellulosic biomass such as wood and agricultural residues
(Balat, 2009a; Cardona and Sánchez, 2007). Current production of
bioethanol is mainly from sugary and starchy feedstock due to their
relatively simpler and easier processing steps. Generally, for sugary
feedstock, sugar is directly fermented by yeasts to produce bioethanol
whereas for starchy materials, an additional hydrolysis steps is
required before fermentation takes place (Cardona and Sánchez,
2007). Hydrolysis is usually catalyzed by cellulose enzymes in order to
breakdown the chains of carbohydrate into glucose which can be
converted into bioethanol by yeasts (Cardona and Sánchez, 2007;
Nigam and Singh, 2011). However, the sustainability of bioethanol
produced from sugary and starchy feedstock is heavily debated due to
their priority for food purposes and large land area is required to
produce the feedstock (Harun et al., 2010).
Alternatively, lignocellulosic biomass appears to be an attractive
alternative feedstock since it is the most abundant renewable biomass
worldwide and does not compete with food crops (Alvira et al., 2010).
The main challenge of using lignocellulosic biomass to produce
bioethanol efficiently is the pre-treatment step. Pre-treatment step is
generally used to break down the complex structure of lignin and to
decrease the fraction of crystalline cellulose to amorphous cellulose
(Cardona and Sánchez, 2007). Hence, accessibility and digestibility of
enzymes (typically cellulases) are enhanced during hydrolysis step
resulting to a higher yield of bioethanol during fermentation process.
Some of the pre-treatment methods are mechanical chipping
grinding, extrusion, alkali or acid pretreatment, ozonolysis, organo-
solv, steam explosion, compressed hot water, ammonia fiber
explosion (AFEX), wet oxidation, microwave pretreatment, ultra-
sound pretreatment and CO2 explosion (Alvira et al., 2010).
Nevertheless, almost all the pre-treatment technologies are costly
and thus, greatly reduce the cost effectiveness of bioethanol
Table 16
Proteins and carbohydrates composition from different microalgae strains (dry matter
basis, %).
production (Cardona and Sánchez, 2007). Furthermore, some of the
pre-treatment methods such as steam explosion, concentrated and
diluted acid can generate toxic compounds (or due to products
degradation) to the environment if no proper waste treatment system
is implemented.
On the other hand, microalgae also appear to be a potential feedstock
for bioethanol production. Despite its high lipid content for biodiesel
production, microalgae also contain carbohydrates (generally not
cellulose) and proteins that can be used as carbon source or substrate
for fermentation (Harun et al., 2010). Table 16 highlights the amount of
carbohydrates and proteins from selected microalgae strains. Different
from terrestrial plants, green and blue-green microalgae are not
lignocellulosic compounds. Therefore, pre-treatment step can be
avoided and consequently simplifies the overall bioethanol production
process. Apart from that, lipid from microalgae can be extracted out for
biodiesel production prior to hydrolysis and fermentation process for
bioethanol production. Furthermore, the remaining microalgae biomass
after fermentation can be utilized in anaerobic digestion process to
produce biomethane. The overall process chain will become more
realistic and sustainable if microalgae is coupled with wastewater
treatment, especially POME treatment in this particular case. Even
though research on fermentation of microalgae biomass to bioethanol is
limited and not yet been commercialized, a lot of advantages are
observed through this process.
5. Conclusion
POME is always regarded as a highly polluting wastewater
generated from palm oil mills. However, reutilization of POME to
generate renewable energies in commercial scale has a great potential
especially when coupled with wastewater treatment process. POME
can be treated anaerobically to breakdown organic matters while
releasing biomethane and sometimes, biohydrogen. Furthermore,
while global interest in culturing microalgae for biodiesel and
bioethanol production is increasing; secondary treated POME has
re-position itself as a good nutrients source for this purpose and
served as a cheap organic fertilizer. This is because POME contains
high amount of nitrate and ortho-phosphate which is required for
microalgae growth. In conclusion, it is an undeniable fact that POME
has its own potential to produce various renewable energies. Such
win–win strategies are in-line with the current zero waste sustain-
ability concept and can make substantial contribution towards better
environmental protection.
Acknowledgement
The authors would like to acknowledge the funding given by
Universiti Sains Malaysia (Short Term Grant No. 304/PJKIMIA/
6039015 and USM Fellowship) for this project.

Microalgae strains Proteins Carbohydrates

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