Chinese Journal of Chemical Engineering 28 (2020) 502–517

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Chinese Journal of Chemical Engineering

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Article

Advances of macroalgae biomass for the third generation of

bioethanol production
Inn Shi Tan 1, Man Kee Lam 2,3,⁎, Henry Chee Yew Foo 1, Steven Lim 4, Keat Teong Lee 5
1
Department of Chemical Engineering, Curtin University, Sarawak Campus CDT 250, 98009 Miri, Sarawak, Malaysia
2
Chemical Engineering Department, Universiti Teknologi PETRONAS, 32610 Seri Iskandar, Perak, Malaysia
3
Centre for Biofuel and Biochemical Research, Institute of Self-Sustainable Building, Universiti Teknologi PETRONAS, 32610 Seri Iskandar, Perak, Malaysia
4
Department of Chemical Engineering, Lee Kong Chian Faculty of Engineering and Science, Universiti Tunku Abdul Rahman, 43000, Selangor, Malaysia
5
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: In recent years, utilization of renewable sources for biofuel production is gaining popularity due to growing
Received 9 January 2019 greenhouse gas (GHG) emissions which causes global warming. There has been a great effort in exploring alter-
Received in revised form 27 May 2019 native feedstock for bioethanol production. In this context, the production of third-generation bioethanol from
Accepted 28 May 2019
macroalgae has emerged as an alternative feedstock to food crop-based starch and lignocellulosic biomass.
Available online 13 July 2019
This is mainly due to the fast growth rate of macroalgae, no competition with agricultural land, high carbohydrate
Keywords:
content and relatively simple processing steps compared to lignocellulosic biomass. This review paper provides
Macroalage an insight of recent innovative approaches for macroalgae bioethanol production, including conventional and ad-
Hydrolysis vanced hydrolysis process to produce fermentable sugar, various fermentation technologies, economic analysis
Biomass and life cycle assessment. With the current technology maturity, efficient utilization of macroalgae as sustainable
Fermentation source for bioethanol and other value-added chemicals production could be achieved in the near future.
Bioethanol © 2019 The Chemical Industry and Engineering Society of China, and Chemical Industry Press Co., Ltd. All rights reserved.

1. Introduction
1.1. The role of biofuels as a sustainable fuel
The world is currently heading towards severe energy crisis due to
rapid growth of world's population and heavy dependency on fossil
fuels in transportation and industrial sectors. The fluctuation of crude
oil price in international market has further worsened the energy secu-
rity particularly among the non-oil producing countries [1]. In 2016, fos-
sil fuels accounted for about 85.5% of the world's primary energy
consumption with 33.3% from crude petroleum oil, 28.1% from coal
and 24.1% from natural gas [2]. The huge dependency on fossil fuels as
energy sources has led to the fast diminishing of this non-renewable
feedstock [3]. Considering the current energy consumption trends, the
supply of petroleum, natural gas and coal will only last for another 45,
60, and 120 years, respectively [4,5]. The decreasing supply of fossil
fuels, especially petroleum has led to the development of innovative
technologies for sustainable production of energy from renewable
sources [6].
Furthermore, continuous utilization of fossil fuels in large quantity
for both development and economic activities has resulted in increasing
⁎ Corresponding auhtor at: Chemical Engineering Department, Universiti Teknologi
PETRONAS, 32610 Seri Iskandar, Perak, Malaysia.
E-mail addresses: tan.s@curtin.edu.my (I.S. Tan), lam.mankee@utp.edu.my (M.K. Lam).
GHG emissions which in turn causing global warming and climate
change effects. At present, the atmospheric CO2 concentration was re-
ported to be around 350 –380 mg ·L -1 and predicted to increase at an
alarming level to 450 mg ·L -1by 2020 if left unchecked [6]. The conse -
quences of high CO2 concentration level in theatmospherecouldlead
to global warming, changing of precipitation patterns, melting of gla-
ciers, frequent occurrence of storms and rising of sea level [7]. The im-
pact of global warming has encouraged relevant industries and
societies to invest in the field of renewable energy and biofuels to miti-
gate the environmental issues.
Renewable energy such as solar energy, wind energy, hydro energy,
and energy from biomass and waste, is defined as energy derived from
regenerative sources and can be replenish over time. It was reported
that utilization of renewable energy could offer a better alternative to
reduce carbon emission and sustain the ecosystem of the Earth [8].
Among all the renewable energy sources, biofuels derived from biomass
has intensively drawn various development endeavors, mainly due to
the wide availability of the feedstock and established biofuels conver-
sion technologies [9]. Biofuels are usually referred to liquid, gas and
solid fuels predominantly produced from biomass. A range of variety
of biofuels can be produced from biomass such as bioethanol,
biomethanol, biodiesel, Fischer–Tropsch diesel, biohydrogen and
biomethane. Since the last decade, global production of biofuel has
been increasing sharply. As shown in Fig. 1, North America had pro-
duced more than 715868 t of oil equivalent per day in 2017 [2]. This

https://doi.org/10.1016/j.cjche.2019.05.012
1004-9541/© 2019 The Chemical Industry and Engineering Society of China, and Chemical Industry Press Co., Ltd. All rights reserved.
 I.S. Tan et al. / Chinese Journal of Chemical Engineering 28 (2020) 502–517 503

800

Biofuel production (×103) tonnes of Oil Equivalent per Day)

North America
700
South & Central America
Europe & Eurasia
600 Asia Pacific

500

400

300

200

100

0
2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 2010 2011 2012 2013 2014 2015 2016 2017

Year

Fig. 1. Latest worldwide biofuel production as of June 2018 [2].

was followed by South and Central America, Europe and Eurasia, and bioethanol added to motor gasoline consumed for transportation in
lastly Asia Pacific. Based on the current biofuel production rate, it is an- the U.S. had increased from about 1.4 billion gallons in 1995 to about
ticipated that the biofuel industries would likely expand in the near fu- 14.4 billion gallons in 2016 [12]. In addition, bioethanol production
ture to address the energy crisis issue and environmental problems. The has the potential to substitute 353 billion gallons of gasoline if
biofuel that is expected to be most widely used around the globe is bioethanol is used as E85 fuel for a middle size vehicle [11]. Since the
bioethanol, with respect to current available production technologies early of 2000s, interest in bioethanol production has been growing
and infrastructure systems [10]. worldwide. It is expected that the demand for biofuels, especially
bioethanol, will increase 3.4 times by 2035 [13]. Fig. 2 shows the total
1.2. Current status of bioethanol production bioethanol production by country or region, from 2007 to 2017 [14,
15]. Global bioethanol fuel production had reached 26.1 billion gallons
According to the United States (U.S.) Department of Energy, 25% of in 2017, with 58.8%, 28.0%, 5.3%, 3.2%, and 1.7% of the production in
the U.S. transportation fuel will be substituted with biofuels by 2040 United States, Brazil, Europe, China, and Canada, respectively. The U.S
[11]. In 2015, the first 88 million gallons of commercial scale cellulosic and Brazil remained as the top two bioethanol producing countries. In
bioethanol capacity was built in the U.S. Today, most of the biofuels 2017, the U.S had produced 15.8 billion gallons of bioethanol while
used in vehicles are blended to gasoline and diesel fuel. The amount of Brazil produced 7.1 billion gallons.

25

20
Bioethanol production (billion gallons)

15 Rest of World
Canada
China
10 Europe
Brazil
USA
5

0
2007 2008 2009 2010 2011 2012 2013 2014 2015 2016
Year

Fig. 2. Bioethanol production by country from 2007 to 2017 [15].

504 I.S. Tan et al. / Chinese Journal of Chemical Engineering 28 (2020) 502–517
The main feedstock of bioethanol production in the U.S is derived
from corn, while for Brazil is mostly based on sugarcane [16]. In fact,
the U.S had utilized 42% of its corn grains (114 million tonnes/year)
for bioethanol production and intended to substitute 10% of its gasoline
demand with bioethanol [17]. Sugarcane-based bioethanol is economi-
cal in Brazil owing to two leading strategies: (1) established directives
to blend gasoline with bioethanol; (2) low sugarcane prices in order
to support the bioethanol industry [18,19]. In other words, these have
drastically reduced the feedstock cost of bioethanol and created positive
development in bioethanol industry. In Brazil, gasoline is necessary to
be blended with bioethanol from 18 to 22%. The percentage of
gasoline–bioethanol blend can be varied from time to time, depending
on the availability of bioethanol [20]. In addition, International Energy
Agency (IEA) also projected that with technology advancement in con-
version of sugarcane to bioethanol, the overall CO2 emissions can be re-
duced by 90% compared to conventional gasoline [21].
1.3. Evolution of bioethanol
First-generation bioethanol (FGB) is being produced commercially
in a number of countries. The term “first-generation bioethanol” or
“conventional bioethanol” refers to bioethanol derived from food
crops (fermentable sugars). Currently, commercial bioethanol is pro-
duced mainly from starch extracted from corn (U.S) and sucrose ex-
tracted from sugarcane (Brazil). Other crops which are rich in starch
or sugar content and used for bioethanol production are potato, sugar
beet, cassava, sweet sorghum, wheat, yam, and barley [22]. Starch can
be easily hydrolysed into glucose; while sucrose, which is a disaccharide
composed of hexoses, fructose and glucose, is readily fermented by
yeast. Fermentation has been carried out for millennia and is a tradi-
tional method for bioethanol production [23]. Several species of yeasts
(e.g. Saccharomyces cerevisiae) metabolize sugars in oxygen free condi-
tions to produce bioethanol and CO2.
However, the production of FGB has received widespread criticism
due to food versus fuel issue. Also, the reduction of GHG emissions
from bioethanol is not as high as expected [24], and the feedstock
could dominate 40% of the bioethanol production cost [23]. In addition,
it has also prompted competition on agricultural land utilization for
food and biofuel production. This could concurrently stimulate many
societal issues, including increasing cost for food sources and disruption
in the food-to-population ratio [25]. For example, total wheat consump-
tion is predicted to attain around 235 Mt. by 2027 as shown in Fig. 3. By
2027, 54% of wheat usage is expected to remain for human food con-
sumption, which is marginally below its present share. Global wheat
utilization as feed is projected to achieve 76 Mt. by 2027, growing at a
Fig. 3. Wheat consumption in developed and developing countries [26].
slower rate than in the past years. On the other hand, wheat usage for
biofuel production will reach 3.0% of global wheat utilization by 2027
compared to 0.9% in the base period.
To overcome these issues, the focus on bioethanol production has
shifted to the development of second-generation bioethanol (SGB) de-
rived from lignocellulosic biomass. Lignocellulosics are usually referred
to waste biomass and mainly obtained from agriculture residues com-
prising of lignin, cellulose and hemicellulose. Although SGB is attractive
due to its nature as non-edible feedstock, the delignification process of
lignocellulosic waste has become a stumbling block for commercial pro-
duction of bioethanol. Different chemical pre-treatment methods with
extreme conditions are used to increase cellulose accessibility to pro-
duce reducing sugar, which lead to the overall process becoming com-
plex and capital intensive [27-31]. Thus, bioethanol production from
lignocellulosic biomass is not sustainable at the present form due to eco-
nomic constraint and negative environmental impacts. Therefore, a
more sustainable feedstock should be identified which can help to over-
come these barriers and improve the sustainability aspects.
In view of these, third-generation bioethanol (TGB) based on
macroalgae biomass has the potential to be a more viable feedstock
over previous generations [32].
1.4. Advantages of bioethanol production from macroalgae
Recently, macroalgae has been gaining wide attention as an alterna-
tive feedstock for bioethanol production [32,33]. Oceans and seas cover
over 70% of the Earth's surface which offer the possibility of sustainable
cultivation of macroalgae biomass feedstock. This is because macroalgae
cultivation does not interfere with agricultural land utilization and does
not require fresh water. Moreover, the growth rate of macroalgae is sig-
nificantly higher than land-based crops [32]. Up to now, there are over
10,000 species of macroalgae reported around the world, with only a
dozen species are being cultivated commercially, while the rest of
macroalgae are exploited from the wild [34]. Nevertheless, over the
last ten years, the world production of macroalgae had been increasing
continuously at an average increment rate of 10% [35]. From Fig. 4, the
brown and red macroalgae were cultivated in much larger quantities
than green macroalgae. In the last few years, the global total production
of aquatic plants (mostly macroalgae and small volume of microalgae)
had dramatically increased and reached 30.1 million tonnes wet weight
in 2016 with a value over USD 11.7 billion [35]. They are largely har-
vested from macroalgae cultivation farms and a small fraction from
wild habitats.
As shown in Table 1, although the amount of macroalgae production
is two times less than that of land-based energy crops, the production of
 I.S. Tan et al. / Chinese Journal of Chemical Engineering 28 (2020) 502–517 505

20000 30000

Total Production (million wet metric tonnes)

18000 Green macroalgae 28000

Production (million wet metric tonnes)

Brown macroalgae
Red macroalgae 26000
16000 Total Production
24000
14000 22000
12000 20000
10000 18000
16000
8000
14000
6000 12000
4000 10000
2000 8000
6000
80 4000
60
40 2000
20
0 0
2003 2004 2005 2006 2007 2008 2009 2010 2011 2012 2013 2014 2015 2016
Year

Fig. 4. World production of farmed macroalgae from 2001 to 2016. Adjusted from [35].

macroalgae is four and six times higher than microalgae and lignocellu-
losic biomass, respectively. Based on current mass cultivation tech-
Table 1 niques, large scale cultivation of macroalgae at sea can sufficiently
World cultivation of macroalgae, microalgae, energy crops, and lignocellulosic biomass
[36]
supply for macroalgae-based biorefinery. The suitable macroalgae spe-
cies that could be integrated in this biorefinery are Laminaria japonica,
Species Group (or phylum) Production/t total/% Eucheuma cottonii (Kappaphycus alvarezii), Undaria pinnatifida, and
①
Macroalgae Gracilaria verrucosa. For red macroalgae species, K. alvarezii, Eucheuma
Eucheuma spp. Red macroalgae 10518771 36.196 spp. and Gracilaria spp. accounted for about 58.7% from the total pro-
Laminaria japonica Brown macroalgae 8219210 28.283
duction of macroalgae. On the other hand, only two brown macroalgae
Gracilaria spp.② Red macroalgae 4149524 14.279
Undaria pinnatifida Brown macroalgae 2069682 7.122 species, U. pinnatifida and L. japonica, accounted for over 35.4% of the
Kappaphycus alvarezii Red macroalgae 1527217 5.255 total macroalgae production. Nevertheless, green macroalgae produc-
Porphyra spp. Red macroalgae 1352520 4.654 tion is low and may be considered negligible, in which Monostroma
Porphyra tenera Red macroalgae 710425 2.445 nitidum and Caulerpa spp. account for only about 0.04% of the total
Eucheuma denticulatum Red macroalgae 273600 0.941
Sargassum fusiforme Brown macroalgae 189910 0.653
macroalgae production.
Phaeophyceae spp Brown macroalgae 33622 0.116 Macroalgae has a wide range of applications and it is being used as
Monostroma nitidum Green macroalgae 7158 0.025 the starting raw material in different sectors such as animal aquaculture,
Codium fragile Green macroalgae 4279 0.015 cosmetics and hydrocolloid industry [44]. Due to current market de-
Enteromorpha clathrata Green macroalgae 3710 0.013
mand and farming techniques, macroalgae-based biorefinery technol-
Caulerpa spp. Green macroalgae 585 0.002
Gracillaria verrucosa② Red macroalgae 450 0.002 ogy should focus on utilizing red and brown macroalgae instead of
Haematococcus pluvialis Green macroalgae 213 0.001 green macroalgae for optimum efficiency. In order to support
Total 29060876 100 macroalgae biomass as the feedstock for biorefinery, intensive coopera-
Microalgae③ tion is essential to improve the ocean-based farming skills among the
Arthrospira sp. Cyanophyta 3000 macroalgae producing countries, such as Indonesia, Malaysia and
Chlorella sp. Chlorophyta 2000 Philippines [6]. One typical example of macroalgae that can be found
Dunaliella salina Chlorophyta 1200 abundantly in South East Asia is E. cottonii (K. alvarezii) which is the
Energy crops④ major commercial source of κ-carrageenan. Goh and Lee [32] had re-
Maize 1134746667 ported that if Eucheuma spp. was used as a feedstock for bioethanol pro-
Palm oil 54384643 duction, the estimated bioethanol yields could reach up to 110,000 t
Rapeseed 76238340
Sugarcane 1841528386
annually. The carbohydrate in macroalgae is composed of two monosac-
Soybean 352643548 charides, which are D-galactose (56.2%) and 3,6-anhydro-galactose
Rice 769657791 (43.8%) [45]. After hydrolysis process, these monosaccharides are suit-
Wheat 771718579 able to be used as substrate during fermentation process for bioethanol
Lignocellulosic biomass⑤ production.
Corn stover 12.6 In addition, macroalgae cultivation does not require arable land and
Switchgrass 9.0 heavy fertilization. Similar to all living plant on the Earth, macroalgae
①
Estimated in wet metric ton and data referred to FAO (2018) [35]. utilize the energy of sunlight to grow via photosynthesis, thus produce
②
Including macroalgae cultured in brackish water. and store organic carbons (carbohydrates). In other words, this is a
③
Estimated in dry metric ton and data referred to Hejazi and Wiffels (2004) [37], promising feedstock that only requires simple inputs (sunlight, sea
Ratledge (2004) [38], Pulz and Gross (2004) [39].
④
Estimated in ton and data referred to FAO (2018) [40,41].
water, nutrient, and carbon dioxide) for growth. These features have en-
⑤
Estimated in dry metric ton a hectare and data referred to Lemus et al. (2002) [41], gendered high optimism for future development in the production of
Shinners and Binversie (2007) [42] [42,43], [38-40]. TGB from macroalgae [32]. The rate of photosynthesis in macroalgae is
506 I.S. Tan et al. / Chinese Journal of Chemical Engineering 28 (2020) 502–517

strongly influenced by thallus morphology and varied based on differ- extracted from the harvested macroalgae through crushing and used
ent species [46]. Table 2 shows the photosynthesis rates of various as biostimulants to improve plant growth rate. For instance, macroalgae
macroalgae. The Enteromorpha sp. (green macroalgae) and Porphyra extracts enhance crop's tolerance towards environmental stress, in-
sp. (red macroalgae) have the highest photosynthesis rates, which are crease yield and quality of agricultural crops. In addition to the afore-
one to two times higher than brown macroalgae. Other advantages of mentioned bio-based products, macroalgae biomass can also be
macroalgae as the feedstock for bioethanol production are low harvest- converted to fermentable sugar through mild acid hydrolysis or enzy-
ing cost and no environmental damage. matic hydrolysis. Macroalgae saccharification/hydrolysis can be
achieved through two different methods: (1) the galactan-cellulose
2. Macroalgae-based Biorefinery Concept mixture are hydrolysed together without separation (direct saccharifi-
cation/hydrolysis) or (2) galactan/carrageenan and cellulose are sepa-
The development of highly efficient biorefinery processes is impor- rated prior to hydrolysis (indirect saccharification/hydrolysis). The
tant for large scale production of bio-based products and bio-energy. resulting hydrolysate is then diverted to fermentation process for
Due to the different chemical structures and composition of macroalgae bioethanol production. Finally, the resulting bioethanol broth can be pu-
compared to terrestrial plants, macroalgae-based biorefinery has a huge rified for product recovery.
potential for the development of new bio-products and bio-energy pro-
duction [6,49]. Macroalgae-based biorefinery embraces the concepts of 2.1. Chemical composition of macroalgae
green and zero pollution. In line with the principle of sustainable devel-
opment, incorporating ecotourism with macroalgae cultivation and re- Since the production efficiency and economic potential of a
fining will be a positive approach to drive this industry towards a biorefinery to produce liquid biofuels are strongly depended on chemi-
sustainable development [32]. Furthermore, this approach can also cal compositions of biomass, thus, a thorough understanding on
bring an alternative livelihood option for the coastal economy. Fig. 5 macroalgae compositions is essential to develop the biorefinery process
shows the conversion of macroalgae into TGB using macroalgae-based [51]. For example, information on the type of sugars (hexose or pen-
biorefinery concept. Nevertheless, a few issues need to be addressed be- tose) found in the medium is important to select suitable fermentation
fore a commercial-scale macroalgae biorefinery can be successfully real- method for bioethanol production. Besides, these information would be
ized [49]. The issues are mainly related to the cultivation and harvesting useful for the selection of biomaterials that can be attained or converted
of macroalgae biomass and also the subsequent processes to convert economically [52].
macroalgae into bio-based products and bio-energy.
As shown in Fig. 5, TGB production from macroalgae involves five 2.1.1. Differences between macroalgae and terrestrial biomass
major consecutive steps: (1) harvest or collection, (2) feedstock han- Macroalgae are significantly different from conventional terrestrial
dling or extraction of the carbohydrates, (3) hydrolysis, (4) fermenta- plant, especially in terms of their chemical composition, physiological
tion, and (5) product recovery. Generally, the macroalgae biomass can and morphological structures [46]. Table 3 shows the differences of car-
be generated either by cultivation and harvesting or by collecting wild bohydrates composition between macroalgae, microalgae, and lignocel-
drift macroalgae. Then, the macroalgae is dewatered for storage purpose lulosic biomass. Fresh macroalgae is mainly composed of moisture and
or to remove any impurities for the downstream bioconversion process. total solid (TS). It is well known that macroalgae biomass has very
Besides, the macroalgae sap (galactan/carrageenan) can be directly high content of moisture, which is typically in the range of 10–90%
(fresh mass) [53]. Volatile solid (VS), which is computed by subtracting
Table 2 ash from total solid (TS) [54], is one of the significant parameters in the
Photosynthesis rate of macroalgae [47] production of bioethanol from macroalgae. Most of the VS of macroalgae
is mainly composed of carbohydrates.
Species① Photosynthesis rate②/μmol·CO2·h−1
Macroalgae have a number of unique compounds, such as carra-
Green macroalgae (Chlorophyceae) geenan, manan, agar, ulvan, laminarin, alginate, fucoidin, and mannitoal
Enteromorpha compressa 316–558 (g dry)−1
Ulothrix speciosa 1092 (g dry)−1
as shown in Table 3 [48,55]. Thus, it is vital to assess their implications
Monostroma grevillei 1466 (g dry)−1 on species selection and associated conversion methods to achieve sus-
Ulva sp. 48.7 (g dry)−1 tainable macroalgae-based biorefinery concept. Compared to terrestrial
Codium fragile 68.3 (g dry)−1 plant, macroalgae do not contain high concentration of starch and lipid
Acrosiphonia centralis 468 (g dry)−1
(except green macroalgae) [63]. In addition, macroalgae are buoyed by
Cladophora rupestris 30.5 (g dry)−1
Enteromorpha sp. 1786 (g wet)−1 water and they do not require woody compound (e.g. lignin) as struc-
tural support [64]. Generally, lignin resists degradation, which is a key
Red macroalgae (Phaeophyceae)
constraint in producing bioethanol from lignocellulosic biomass. Since
Iridaea cordata 29.4 (g dry)−1
Porphyra sp. 1808.7 (g dry)−1 macroalgae have low lignin content, this offers suitability for down-
Asparagopsis taxiformis 174 (g dry)−1 stream processing without costly additional pre-treatment such as
Chondrus crispus 21.2 (g dry)−1 delignification and removal of lignin-based inhibiting compounds [65].
Dictyopteris australis 324 (g dry)−1 Macroalgae usually contain less than 5 wt% of lipid, which is differ-
Dictyopteris justii 118 (g dry)−1
Delesseria sanguinea 37.9 (g dry)−1
ent from microalgae that have high lipid content (10%–20% dry mass)
Gracilaria sp. 85 (g wet)−1 [36,66,67]. Due to their low lipid content, the production of biofuel
from macroalgae is expected to depend on carbohydrates content
Brown macroalgae (Rhodophyceae)
rather than lipid content. Macroalgae generally have higher ash content
Alaria marginata 109.3 (g dry)−1
Laminaria sp. 124 (g dry)−1 (20.8%–89.7% dry mass) and alkali metal content (10%–50% dry mass)
Macrocystis sp. 171.8 (g dry)−1 compared to terrestrial biomass [68,69]. On the other hand, macroalgae
Ceramium nitens 867 (g dry)−1 have lower protein content (7%–15% dry mass) [70] compared to most
Cymathere triplicata 58.5 (g dry)−1
microalgae which have higher content of protein (40%–60% dry mass)
Dictyopteris sp. 221 (g dry)−1
Fucus sp. 561 (g dry)−1 [67].
Sargassum sp. 415 (g dry)−1
①
Data referred to Lobban and Wynne, (1981) [48] and Gao and McKinley, (1994) [47].
2.1.2. Carbohydrate composition of macroalgae
②
Estimated in μmol CO2·h−1 on the basis of dry mass (g dry−1) or wet mass ((g Macroalgae usually contain high concentration of carbohydrate
wet)−1) [43], [37]. within their biomass [71]. The carbohydrate can be separated into
 I.S. Tan et al. / Chinese Journal of Chemical Engineering 28 (2020) 502–517 507

Macroalgae
cultivation
Feedstock processing
Harvest or (cleaning, dewatering, Liquid waste and
collection drying, crushing, slurrying, debris
etc)
Wild
macroalgae

Hot
Direct Macroalgae
water/alkali
hydrolysis powder
extraction
Purification and
separation

Acid/enzymatic Indirect
hydrolysis hydrolysis

Cellulose Carrageenan/ Proteins

galactan
Reducing
sugars

Pretreatment
Acid/enzymatic Pharmaceutical
and enzymatic
hydrolysis products
hydrolysis
Reducing sugars
Reducing sugars
Fermentation

Purification

Third-generation
bioethanol

Fig. 5. Schematic flow diagram of conversion of macroalgae into third-generation bioethanol (TGB) using macroalgae biorefineries concept [50].

soluble and insoluble compounds through chemical method [71,72].
The carbohydrate contents in red, brown, and green macroalgae are
usually ranged from 30% to 60%, 30%–50%, and 25%–50% dry mass, re-
spectively [67,68,70,73,74]. Since the composition of carbohydrates in
macroalgae varies widely among different species, it is necessary to
ascertain their carbohydrate compositions in order to produce
bioethanol and other related bio-products efficiently.
2.1.2.1. Red macroalgae. Red macroalgae generally consists of cellulose,
galactan, and glucan. The cell wall of macroalgae accounts for up to

Table 3
Carbohydrate composition of macroalgae, microalgae, and lignocellulosic biomass

Macroalgae① Microalgae② Lignocellulosic biomass③

Green macroalgae Red macroalgae Brown macroalgae

Polysaccharide Polysaccharide Polysaccharide Polysaccharide Polysaccharide

Mannan Carrageenan Laminarin Starch Cellulose
Ulvan Agar Mannitol Monosaccharide Hemicellulose
Starch Cellulose Alginate Arabinose Lignin
Cellulose Lignin Fucoidin Fucose Starch
Monosaccharide Monosaccharide Cellulose Galactose Monosaccharide
Glucose Glucose Monosaccharide Glucose Glucose
Mannose Galactose Glucose Mannose Xylose
Rhamnose Agarose Galactose Rhamnose
Xylose Fucose Ribose
Xylose Xylose
①
Data referred to Borines et al., (2013) [33], Trivedi et al. (2015b) [43] and Yun et al., (2016) [56].
②
Data referred to Ho et al., (2014) [57,58].
③
Data referred to Paulova et al., 2015 [59] and van Kuijk et al., (2015) [56,58-62].
508 I.S. Tan et al. / Chinese Journal of Chemical Engineering 28 (2020) 502–517

65 wt% of dry matter and comprises three domains: fibrillar wall, amor-
phous matrix and glycoprotein domain [75]. The amorphous matrix of
red macroalgae is dominated with sulphated galactans, such as agar
and carrageenan, which are long-chain polysaccharides that are impor-
tant for their gel-forming abilities [76-79]. The main gelling portion in
agar is agarose, which consists of D-galactose and 3, 6-anhydro-L-
aglactose (AHG) with alternate α-1, 3-and β-1, 4-linkages [73]. Agar
can be easily hydrolysed to produce galactose monomers [80], whereas
carrageenan is a sulphated polygalactan composed of D-galactose and 3,
6-anhydro-D-galactose units with a 15%–49% ester-sulfate content [81].
Carrageenan can be further classified as lambda (λ), kappa (κ), and iota
(ι), based on their gel-forming ability [82]. Carrageenan can be ex-
tracted from red macroalgae by dissolving it into aqueous solution.
Palmaria palmata contains the highest carrageenan with the concentra-
tion of 354 mg·g−1 and comparison of bioethanol yield for 20 varieties
of macroalgae show that P. palmata is the best suited for the large-scale
production of bioethanol [83].
2.1.2.2. Brown macroalgae. The primary carbohydrates in brown
macroalgae may consist of 55% dry mass of laminarin and mannitol car-
bohydrate [84,85]. Laminarin is a β-1, 3-linked glucan with either man-
nitol (M-chains) or glucose (G-chain) attached to the reducing end [86].
Brown macroalgae can be hydrolysed by laminarase (endo-1, 3 (4)-β-
glucanase) to produce glucose monomers [87]. Mannitol has osmoregu-
latory properties and it can be derived from D-mannose (six carbon
sugars). Besides, mannitol is a sugar alcohol that can be easily fermented
into bioethanol [88]. Brown macroalgae also contains mainly alginate
and cellulose in their cell wall, which are two structural polysaccharides
acting as structural support to prevent ruptures during currents and
tidal fluctuations [89].
2.1.2.3. Green macroalgae. Green macroalgae such as Valonia sp. is rich in
cellulose (up to 70 wt%) and pectin as the main structural polysaccha-
rides in the cell wall [90,91]. In addition, green macroalgae consists of
carbohydrate in the form of starch as food reserves [92]. It consists of
polymerized glucose molecules and deposited in chloroplasts in the
form of gains [93]. Green macroalgae also contains sucrose and other
carbohydrates such as ulvan [94,95]. Ulvan is a water-soluble sulphated
polysaccharide found in Ulva sp. (up to 29% dry mass) [94]. The main re-
peating disaccharide units reported are ulvanobiouronic acid 3-sulfate
types containing either glucuronic or iduronic acid [96]. In addition,
minor repeat units had been reported which contained sulfated xylose
replacing the uronic acid or glucuronic acid as a branch on O-2 of the
rhamnose-3-sulfate [97].
2.2. Hydrolysis of macroalgae biomass
Saccharification/hydrolysis is an important stage prior to bioethanol
production, in which complex polysaccharides are hydrolysed into sim-
ple reducing sugars. Unlike lignocellulosic waste such as wood and
grass, macroalgae contains no lignin and thus, susceptible to the hydro-
lysis process with no interference with the catalyst.
2.2.1. Acid hydrolysis
Acid hydrolysis uses acids such as sulfuric acid (H2SO4) to hydrolyze
glucosidic bonds. For effective acid hydrolysis of macroalgae biomass,
solid/liquid (S/L) ratio, acid type, acid concentration, reaction time,
and reaction temperature are important parameters to be optimized.
Up to now, many studies had been conducted to hydrolyze carbohy-
drates in macroalgae biomass, such as Gelidium amansii and
K. alvarezii (E. cottonii) via acidic method. Galactose components in
G. amansii and K. alvarezii are mainly polysaccharides κ-carrageenan
and agar, respectively. The comparison of acid hydrolysis from various
macroalgae is shown in Table 4. In general, reducing sugar such as glu-
cose and galactose was produced from red macroalgae.
Dilute sulfuric acid was used as catalyst to treat red macroalgae,
G. amansii and K. alvarezii biomass with 0.1–0.94 mol·L-1 H2SO4 at 100
to150 °C for 0.25 to 24 h [76,79,98,99,101-103]. In order to attain
bioethanol with high concentration, sugars in the acid hydrolysate of
K. alvarezii was concentrated by evaporation process. On the other hand,
two-step semi-continuous hydrolysis process is recommended to reduce
the processing cost of hydrolysis process. For example, K. alvarezii was
first hydrolyzed with 0.45 mol·L-1 H2SO4 and the hydrolysate were
recycled to be used in second hydrolysis step [103]. Through this method,
the total reducing sugar produced was 20.6 g·L−1 and the process was able
to be repeated for five times without significant reduction in efficiency.
Hargreaves et al. [98] had reported on production of reducing sugars
from K. alvarezii. The hydrolysis process was carried out with high bio-
mass loading of about 33.33 wt% raw material, 0.18 mol·L-1 H2SO4 at
121 °C for 1 h and resulted in a hydrolysate containing optimum galac-
tose concentration of 81.62 g·L−1. However, about 20.7 g·L−1 of by-
product 5-hydromethylfurfural (5-HMF) was also produced. This
showed that sugar decomposition occurred and 5-HMF was an inhibitor
during the fermentation process. In general, the amounts of 5-HMF

Table 4
Comparison of acid hydrolysis from red and brown macroalgae [98] [99] [100]

Macroalgae Conditions for acid hydrolysis Sugars Yield of sugar/ Concentration of References
material produced g sugar ·(g macroalgae)-1 sugar/g·L−1

Red macroalgae
Gelidium amansii 10% (w/v) biomass loading, 0.1 mol·L-1 H2SO4 at 150 °C for 15 min Glucose 0.05 5.0 [76]
Galactose 0.30 30
Gelidium amansii 10% (w/v) biomass loading, 0.2 mol·L-1 H2SO4 at 121 °C for 59 min Glucose 0.02 2 [79]
Galactose 0.22 22
-1
Kappaphycus alvarezii 10% (w/v) biomass loading, 0.2 mol·L H2SO4 at 130 °C for 15 min Glucose 0.049 4.9 [101]
Galactose 0.256 25.6
Kappaphycus alvarezii 10% (w/v) biomass loading, 0.2 mol·L-1 H2SO4 at 130 °C for 15 min Glucose 0.0089 0.89 [102]
Galactose 0.2387 23.87
Kappaphycus alvarezii Repeated acid hydrolysis Reducing sugars 0.262 20.6 [103]
5% (w/v) biomass loading, 0.45 mol·L-1 H2SO4 at 100 °C for 1 h
(5 cycles, overall 250 g sampled used)
Kappaphycus alvarezii 33.33% (w/w) biomass loading, 0.18 mol·L-1 H2SO4 at 121 °C for 1 h Galactose 0.51 81.62 [98]
Kappaphycus alvarezii 20% (w/v) biomass loading, 0.2 mol·L-1 H2SO4 at 110 °C for 90 min Glucose 0.03 2.61 [99]
Galactose 0.126 10.11
Eucheuma denticulatum 6.15% (w/w) biomass loading, 0.1 mol·L-1 H2SO4 at 160 °C Glucose 0.75 51.47 [104]

Brown macroalgae
Undaria pinnatifida 5% (w/v) biomass loading, 0.94 mol·L-1 H2SO4 at 120 °C for 24 h Glucose 0.065 3.3 [100]
Xylose 0.002 0.1
Fructose 0.004 0.2
 I.S. Tan et al. / Chinese Journal of Chemical Engineering 28 (2020) 502–517 509

produced are highly depended on hydrolysis conditions. Severe hydro-
lysis conditions such as high concentration of acid will favor the produc-
tion of 5-HMF and decrease the reducing sugars production. In addition,
brown macroalgae had been used to produce reducing sugars under
high acid concentration. Lee et al. [100] applied acid hydrolysis to
U. pinnatifida by using 0.94 mol·L-1 H2SO4 at 120 °C for 24 h. Several
types of reducing sugars were produced, such as glucose, xylose, and
fructose, but in low concentration. In addition, high energy was required
in this hydrolysis method due to exceptionally long reaction time. Fur-
thermore, using concentrated acid in hydrolysis process also suffers
from several problems such as waste treatment of the acid residue, reac-
tor corrosion, and producing huge amount of waste [105].
In addition, dilute sulfuric acid hydrolysis with microwave-assisted
heating could be considered a promising alternative conversion route
for sugar recovery. Teh et al. [104] used red macroalgae Eucheuma
denticulatum, which was subjected to microwave heating with a fre-
quency of 2.45 GHz and power maintained at 10 A. They found that
the highest total reducing sugars were 51.47 g·L−1 along with a low
by-product 5-HMF of 0.20 g·L−1, when the biomass was treated
under the optimum conditions at 160 °C with 0.1 mol·L-1 H2SO4.
2.2.2. Enzymatic hydrolysis
Besides acid hydrolysis, enzymatic hydrolysis is an alternative
method to hydrolyze macroalgae biomass. In addition, enzymatic hy-
drolysis is a more effective approach for producing sugars from cellu-
losic biomass because of the higher conversion yield and the
production of a lower amount of toxic hydrolysates compared to that
generated by the acid hydrolysis process [106]. However, it should be
noted that specific enzyme is usually tailored to certain types of polysac-
charide while macroalgae is usually composed of various types of poly-
saccharides [107].
Cellulose, one of the components of macroalgae, could be effectively
hydrolysed with the help from enzymes. Several studies had reported
on enzymatic hydrolysis of macroalgae biomass by using cellulase and
cellobiase (β-glucosidase), which were the same enzymes used to hy-
drolyse lignocellulosic biomass [99,108,109]. Through enzymatic hy-
drolysis, cellulose was converted to reducing sugars by the cellulase
enzyme to produce bioethanol via fermentation process. The mecha-
nism of cellulolytic enzymes follows three important steps: (1) adsorp-
tion, (2) biodegradation, and (3) desorption [110]. Several applications
of enzyme on macroalgae are summarized in Table 5.
Enzymatic hydrolysis using meicelase alone (derived from
Trichoderma viride) was sufficient to complete the saccharification of
Ulva pertusa (green macroalgae) and Alaria crassifolia (brown
macroalgae), primarily due to only glucan (single polysaccharide) was
presence in both macroalgae [109]. Green macroalgae Ulva fasciata
Delile with carbohydrate content of (43 ± 4.5)% dry weight had been
successfully used to produce glucose by enzymatic hydrolysis with
18.4% hydrolysis efficiency.
The enzymatic hydrolysis of cellulose residue that obtained from red
macroalgae K. alvarezii after carrageenan extraction showed 84% of en-
zymatic hydrolysis efficiency [98]. The study also reported that the low
efficiency of enzymatic hydrolysis over biomass concentration were
caused by diffusional limitation and high solid concentrations that
prohibited the action of the cellulolytic enzymes.
Another enzyme, β-glucosidase or also known as Novozyme 188, is
commonly used simultaneously with cellulase enzyme as a supplement
enzyme. Recently, it had been used to produce bioethanol from
G. verrucosa (red macroalgae) with hydrolysis efficiency of 88%. The op-
timal hydrolysis rate of (1.08 ± 0.02) g·L−1·h−1 was attained after 36 h
and was exceptionally higher than the hydrolysis efficiency of other
macroalgae. This could be due to the high carbohydrate content in the
macroalgae biomass and optimized enzymatic hydrolysis conditions
[111]. The study also revealed that each enzyme had its own character-
istics and could only perform well under their respective optimum
conditions.
Several important factors that influence enzymatic hydrolysis effi-
ciency are pH, temperature, substrate and enzyme loading [112]. Be-
sides, macroalgae dedicated enzymes such as laminarinase and
agarose were also employed to saccharify macroalgae [113]. Neverthe-
less, these enzymes showed lower performance which indicated that
pretreatment of macroalgae biomass prior to enzymatic hydrolysis
was essential.
2.3. Chemical pre-treatment of macroalgae for recovery of cellulose
The establishment of an efficient pre-treatment method is to facili-
tate the conversion of sugars during the hydrolysis process and subse-
quently to increase bioethanol yield [114,115]. The goal of the pre-
treatment process is to reduce the crystallinity of cellulose and increase
the porosity of the cellulosic materials, so that cell wall-bound carbohy-
drates are accessible for hydrolysis process [116-118].
Most researchers had concluded that high sugar recovery during
pre-treatment step could be attained by optimizing biomass loading,
pre-treatment time, temperature and acid concentration. However, it
should be noted that unfavorable condition of pre-treatment and hy-
drolysis could further convert the reducing sugars into undesirable by-
products such as formic acid, acetic acid and furanic compounds. Several
examples of bioethanol production and pre-treatment methods for
macroalgae are described in Table 6.
Generally, acid hydrolysis can either serve as pre-treatment step
before enzymatic hydrolysis or as the actual chemical method to

Table 5
Comparison of enzymatic hydrolysis from various macroalgae feedstocks

Macroalgae material Conditions for enzymatic hydrolysis Sugars Yield of sugar/g sugar References
produced ·(g macroalgae)-1

Red macroalgae
Gracilaria verrucosa 10% (w/v) biomass loading, 20 FPU cellulase·g−1 biomass, 60 CBU β-glucosidase·g−1 biomass, 0.05 Glucose 0.876 [111]
(left over pulp after mol·L-1 citrate phosphate buffer, 50 °C, pH 5.0, 150 r·min-1 for 36 h
agar extraction)
Kappaphycus alvarezii 7.32% (w/v) biomass loading, 45 FPU cellulase·g−1 biomass, pH 5, 50 °C, 150 r·min-1 for 24 h Glucose 0.841 [98]
(cellulose residue)

Brown macroalgae
Alaria crassifolia 30% (w/v) biomass loading, 5 g·L−1 of Meicelase (cellulase ~73.3 units·g−1 and cellobiase~227 Glucose 0.224 [109]
units·g−1), 0.1 mol·L-1 citric acid, pH 5.5, 50 °C, 100 r·min-1 for 120 h

Green macroalgae
Ulva pertusa 30% (w/v) biomass loading, 5 g·L−1 of Meicelase (cellulase ~73.3 units·g−1 and cellobiase~227 Glucose 0.143 [109]
units·g−1), 0.1 mol·L-1 citric acid, pH 5.5, 50 °C, 100 r·min-1 for 120 h
Ulva fasciata Delile 5% (w/v) biomass loading, 2% w/v cellulase, pH 4.8, 45 °C, 150 r·min-1 for 36 h Glucose 0.184 [74]
510 I.S. Tan et al. / Chinese Journal of Chemical Engineering 28 (2020) 502–517

Table 6
Comparison of pretreatment of various macroalgae feedstocks

Macroalgae material Hydrolysis method Yield References

of
sugar
/%

Laminaria japonica Acid pretreatment/enzyme hydrolysis 26.80 [108]

(floating residue from
alginate industry)
1. Biomass loading (w/v)① in 0.1% (w/v) H2SO4, 121 °C for 1 h
2. 2% (w/v) biomass loading, 45 FPU cellulase·g−1 biomass, 55 CBU cellobiase·g−1 biomass, 50 °C, 150 r·min-1, pH 5.5 for
24 h
Laminaria japonica Acid pretreatment/enzyme hydrolysis 83.37 [119]
(whole biomass)

Laminaria japonica
(whole biomass)
1. 18% (w/v) biomass loading in 0.06% (w/v) H2SO4, 170 °C for 15 min
2. 0.5% (w/v) biomass loading, 15 FPU cellulase·g−1 biomass, 70 pNPGU β-glucosidase·g−1 biomass, 50 °C, 120 r·min-1,
pH 4.8 for 24 h
Acid pretreatment/enzyme hydrolysis 72.40 [77]

1. 0.1% (w/v) biomass loading in 0.15 mol·L-1 H2SO4, 121 °C for 15 min
2. biomass loading①, Cellulast 1.5 L, Viscozyme, 50 °C, 100 r·min-1, pH 5.5 for 24 h
Saccharina japonica Acid pretreatment/enzyme hydrolysis 89.38 [120]

1. Biomass loading① in 0.30% (w/v) H2SO4, 147.14 °C for 27.85 min

2. Biomass loading①, 15 FPU cellulase·g−1 biomass, 70 pNPGU β-glucosidase·g−1 biomass, 50 °C, 150 r·min-1, pH 4.8 for
24 h
Sargassum fulvellum Acid pretreatment/enzyme hydrolysis 73.30 [77]

1. 0.1% (w/v) biomass loading in 0.15 mol·L-1 H2SO4, 121 °C for 15 min
2. biomass loading①, Cellulast 1.5 L, Viscozyme, 50 °C, 100r·min-1, pH 5.5 for 24 h
Alaria crassifolia Acid pretreatment/enzyme hydrolysis 41 [109]

1. 33.33% (w/v) biomass loading in 0.2 mol·L-1 H2SO4, 121 °C for 30 min
30% (w/v) biomass loading, 5 g·L−1 of Meicelase (cellulase~73.3 units·g−1 and cellobiase~227 units·g−1), 0.1 mol·L-1
citric acid, pH 5.5, 50 °C, 100 r·min-1 for 120 h
Gelidium amansii Acid pretreatment/enzyme hydrolysis 24.2 [77]

1. 0.1% (w/v) biomass loading in 0.15 mol·L-1 H2SO4, 121 °C for 15 min
2. biomass loading①, Cellulast 1.5 L, Viscozyme, 50 °C, 100 r·min-1, pH 5.5 for 24 h
Ulva pertusa Acid pretreatment/enzyme hydrolysis 26.3 [109]

1. 33.33% (w/v) biomass loading in 0.2 mol·L-1 H2SO4, 121 °C for 30 min
2. 30% (w/v) biomass loading, 5 g·L−1 of Meicelase (cellulase ~73.3 units·g−1 and cellobiase~227 units·g−1), 0.1 mol·L-1
citric acid, pH 5.5, 50 °C, 100 r·min-1 for 120 h
Ulva lactuca Acid pretreatment/enzyme hydrolysis 35.7 [77]

1. 0.1% (w/v) biomass loading in 0.15 mol·L-1 H2SO4, 121 °C for 15 min
2. biomass loading①, Cellulast 1.5 L, Viscozyme, 50 °C, 100 r·min-1, pH 5.5 for 24 h
①
Biomass loading: data was not reported.

hydrolyze biomass to produce reducing sugars [106]. Dilute sulfuric
acid (H2SO4) is commonly used for pre-treatment of macroalgae bio-
mass. An investigation on H2 SO 4 pre-treatment of two types of
macroalgae, U. pertusa and A. crassifolia was carried out by
Yanagisawa et al. [109]. These two types of macroalgae were
pretreated at 0.2 mol·L-1 H2SO4, 121 °C for 30 min followed by enzy-
matic hydrolysis. According to this study, for both macroalgae, the
glucose concentration after the pretreatment process was about 1.8
times higher than the enzymatic hydrolysis of macroalgae biomass
without pretreatment.
Ge et al. [108] reported that macroalgae floating residue wastes from
L. japonica alginate industry could be effectively converted to simple
sugars using dilute H2SO4 pre-treatment and followed by enzymatic hy-
drolysis consisting of cellulase and cellobiose. The research found that
the floating residues were promising feedstock for bioethanol produc-
tion due to their high amount of cellulose content with low hemicellu-
lose and lignin concentration. Under extremely low acid concentration
(ELA) pre-treatment (0.06% H2SO4) applied to L. japonica, it was found
that the maximum glucan content of 29.09% was attained, which was
four-fold higher than that of the untreated L. japonica [119]. Further
study done by Lee and Kim [120] with similar dilute H2SO4 pre-
treatment conditions (0.30% H2SO4 at 147.14 °C for 27.85 min) on
Saccharina japonica showed an improvement in the glucan content
and enzymatic efficiency compared to untreated S. japonica, by 4.7-
and 2.6-fold increment, respectively. Nevertheless, the main disadvan-
tage of pre-treatment using acid hydrolysis is the formation of multiple
inhibitory by-products which is probably due to cell disruption and in-
hibitory compounds generated during the acid hydrolysis conditions
[85]. These inhibitory compounds such as furfural, 5-
hydroxymethylfurfural (5-HMF), levulinic acid, and caffeic acid are de-
rived primarily from reducing sugar such as xylose and galactose in
macroalgae biomass [65]. The inhibitory compounds can be removed
by using lime and activated carbon but may increase the bioethanol pro-
duction cost [98].
Although H2SO4 is a powerful agent for cellulose pre-treatment, it is
toxic, corrosive, and hazardous [121]. Therefore, highly corrosive resis-
tant reactor is usually required which indirectly increases the
bioethanol production cost. In addition, it is also very difficult to recycle
 I.S. Tan et al. / Chinese Journal of Chemical Engineering 28 (2020) 502–517 511

H2SO4 and its disposal would require proper waste water treatment cellulose (45 mg) under reaction conditions of 150 °C for 24 h with
facilities. 50 mg of Amberlyst (TM) 15 [133].
Literatures on investigating the sugar recovery from macroalgae by
using solid acid catalyst were still very limited. In general, all types of
2.4. Solid acid catalyst hydrolysis and pretreatment techniques
carrageenan and agar that are extracted from macroalgae are soluble
in hot water at temperatures above its gel melting temperature. Thus,
Chemical hydrolysis and pre-treatment techniques using liquid acid
it is possible to hydrolysis carrageenan or agar by using solid acid cata-
catalyst are preferred in terms of high hydrolysis efficiency and mass
lyst. Tan et al. [135] had successfully demonstrated the feasibility to hy-
transfer rate. Nevertheless, this method also faces several limitations,
drolyze carbohydrates from E. cottonii extract to simple reducing sugar
such as intensive product separation, potential corrosion on reactor
prior to fermentation process by using Amberlyst (TM) 15 and the opti-
wall, difficult to recover the catalyst and additional waste treatment
mum sugar yield attained was 39.7%. Recently, Jeong et al. [134] re-
step is required. As an alternative to dilute acid hydrolysis and pre-
ported that a 61 g·L−1 total reducing sugars was obtained from
treatment method, it is also possible to use solid acid catalyst which
G. verrucosa with 15% Amberlyst (TM) 15 at 140 °C, and 150 min condi-
can overcome these drawbacks [112]. Recent advancement in biomass
tions. In addition, Dowex (TM) Dr-G8 was explored as a potential cata-
pre-treatment and hydrolysis with solid acid catalyst has shown prom-
lyst to hydrolyze carbohydrates from E. cottonii or macroalgae extract
ising replacement for liquid acid hydrolysis and pre-treatment [123]. In
[137]. The effects of catalyst loading (2%–10% w/v), reaction tempera-
order to maintain the process efficiency, the solid acid catalysts should
ture (110–140 °C) and biomass loading (8%–20%) on the simple reduc-
have a high number of Brӧnsted acid sites, good affinity for the reactant
ing sugar production were studied. The highest values of galactose yield
substrates, high surface area and good thermal stability [124]. Other im-
were obtained at 43.2% in E. cottonii and 49.4% in macroalgae extract
portant characteristics of solid acids such as high catalyst porosity, ded-
when treated under optimum reaction conditions (catalyst loading:
icated functional groups and high stability in water are important in the
6% w/v (E. cottonii) and 8% w/v (macroalgae extract), temperature 120
biomass hydrolysis process [125].
°C, 1 h of reaction time and 16% biomass loading).

2.4.1. Hydrolysis with solid acid catalyst
The hydrolysis mechanism of water soluble polysaccharide consists
of the following steps: (1) the soluble polysaccharide molecules diffuse
onto the surface acidic sites of the solid acid catalyst; (2) β-1, 4 glyco-
sidic bonds access to the acidic sites; and (3) these bonds are being
cleaved randomly, and the polysaccharides are hydrolysed to reducing
sugars e.g. glucose and galactose. In the existing literature, four different
classes of solid catalysts were reported: (1) cation-exchange resins,
(2) siliceous-based materials, (3) metal oxides, and (4) sulfonated
amorphous carbons; in which all these solid acid catalysts had equiva-
lent or stronger acidity compared to liquid acid [126].
Recently, several solid acids were reported to have high catalytic ac-
tivity to hydrolyze cellulose, starch and other polysaccharides [127-
131]. These suggested that direct hydrolysis of biomass using solid
acid catalyst without subsequent enzymatic hydrolysis is possible.
Strong acid cation-exchange resins bearing sulfonic acid sites were
found to be particularly effective in the hydrolysis of cellulose. An
added advantage of cation-exchange resins is the simultaneous removal
of inhibitors by the resins itself during the hydrolysis process [132].
Several applications of solid acid catalyst on various substrates are
summarized in Table 7. Amberlyst (TM) 15 and Nafion resins were the
most popular solid catalysts in organic synthesis process, mainly due
to their high selectivity in non-aqueous and aqueous solution [126]. In
fact, resins are both chemically and thermally stable (up to 280 °C).
Amberlyst (TM) 15 is an effective catalyst to selectively convert cellu-
lose to glucose. About 25% of glucose yield was attained from milled
2.4.2. Pre-treatment with solid acid catalyst
Unlike hydrolysis, during the pre-treatment process, reducing
sugars are less likely to form because it is generally conducted at mild
reaction conditions, such as low reaction temperature, short processing
time, and low acid concentration [138]. After pre-treatment process,
pre-treated biomass will be further hydrolysed by either enzymatic or
chemical method as discussed in Section 2.2. Pre-treatment mediated
with solid acid catalysts is superior than conventional liquid catalyst, es-
pecially in the destruction of biomass with fewer side-reactions occur-
ring [139]. As the pre-treatment process proceeds, part of the cellulose
is converted into oligosaccharide and the structure of biomass becomes
more irregular and disordered, resulting in low crystallinity of cellulose
or an amorphous structure. Thus, the resulting cellulose becomes more
susceptible to enzymatic hydrolysis in the subsequent enzymatic
process.
Recently, a novel process to treat lignocellulosic biomass using solid
acid catalysts had been investigated, and the influence of pre-treatment
conditions such as pre-treatment temperature, pre-treatment time, and
types of catalyst on the efficiency of subsequent enzymatic hydrolysis
was evaluated [140]. The process used rice straw as feedstock and
Amberlyst 35 Dry, sulphated zirconia (SA-J1), and a novel sulfonated
mesoporous silica catalyst (MPS-1) as catalysts (Table 8). The pre-
treatment process was carried out in an autoclave batch reactor at
110–180 °C for 3 h. After the pre-treatment, there was no significant dif-
ference among the liquefaction rates of the pre-treated rice straw using
these four catalysts. However, pre-treatment using SA-J1 showed higher

Table 7
Comparison of solid acid hydrolysis from various substrates

Substrate Catalyst Conditions for solid acid hydrolysis Glucose References

yield/%

Cellulose Nafion-50 0.5% (w/v) of cellulose, 160 °C, 4 h, and 1% (w/v) of Nafion-50 35 [60]
Cellulose Amberlyst (TM) 22.5% (w/v) of cellulose, 150 °C, 24 h, and 1% (w/v) of Amberlyst (TM) 15 25 [133]
15
Cellulose H-ZSM5 1% (w/v) of cellulose, 150 °C, 24 h, and 0.9% (w/v) of H-ZSM5 24 [61]
Gracilaria verrucosa Amberlyst (TM) 13.33% (w/v) of Gracilaria verrucosa, 140 °C, and 2.5 h, and 15% (w/v) of Amberlyst 51.9 [134]
15 (TM) 15
Cellobiose HSM-5B 0.2% (w/v) of cellobiose, 175 °C, 0.5 h, and 1% (w/v) of HSM-5B 33.7 [62]
E. cottonii Amberlyst (TM) 12.5% (w/v) of E. cottonii, 120 °C, 1.5 h, and 4% (w/v) of Amberlyst (TM) 15 39.7 [135]
15
E. cottonii Dowex (TM) 16% E. cottonii, 120 °C, 1 h, and 6% (w/v) of Dowex (TM) Dr-G8 43.2 [136]
Dr-G8
Macroalgae extract (Extracted from E. Dowex (TM) 16% E. cottonii, 120 °C, 1 h, and 8% (w/v) of Dowex (TM) Dr-G8 49.4 [136]
cottonii) Dr-G8
512 I.S. Tan et al. / Chinese Journal of Chemical Engineering 28 (2020) 502–517

Table 8
Comparison of solid acid pretreatment of various substrates

Substrate Hydrolysis method Yield of References

sugar/%

Rice straw Solid acid pretreatment/enzyme hydrolysis 76.20 [140]

1. 18% (w/v) biomass loading in 2% (w/v) Amberlyst 35 Dry, 150 °C for 3 h.

2. 2% (w/v) biomass loading, 45 FPU cellulase·g−1 biomass, 45 °C, 150 r·min-1, pH 5.5 for 72 h
Rice straw Solid acid pretreatment/enzyme hydrolysis 83.20 [140]

1. 18% (w/v) biomass loading in 2% (w/v) sulphated zirconia, 150 °C for 3 h.

2. 2% (w/v) biomass loading, 45 FPU cellulase·g−1 biomass, 45 °C, 150 r·min-1, pH 5.5 for 72 h.
Rice straw Solid acid pretreatment/enzyme hydrolysis 62.40 [140]

1. 18% (w/v) biomass loading in 2% (w/v) mesoporous silica catalyst, 150 °C for 3 h.
2. 2% (w/v) biomass loading, 45 FPU cellulase·g−1 biomass, 45 °C, 150 r·min-1, pH 5.5 for 72 h
Macroalgae cellulosic Solid acid pretreatment/enzyme hydrolysis 99.80 [136]
residue

1. 10% (w/v) biomass loading in 4% (w/v) Dowex (TM) Dr-G8, 120 °C for 30 min.
2. 2% (w/v) biomass loading, 45 FPU cellulase·g−1 biomass, 52 CBU·g−1 β-glucosidase, 50 °C, 150 r·min-1, pH 4.8,
120 r·min-1 for 30 h.

yields of oligosaccharide after pre-treatment compared to MPS-1 and
Amberlyst 35 Dry catalyst. In contrast, MPS-1 and Amberlyst 35 Dry
had higher yields of monosaccharide (xylose and glucose) than SA-J1
catalyst. These results indicated the possibility of a direct saccharifica-
tion of lignocellulosic biomass using a strong solid acid catalyst (e.g.
Brӧnsted-type) without subsequent enzymatic hydrolysis.
Li et al. [139] indicated that the highest total reducing sugar (TRS)
yield of 66.7% was achieved using the mesocarbon microbeads-based
solid acid at 140 °C for 4 h. This work offers a solid catalyst that exhibited
excelled recyclability, regeneration, and catalytic ability for the conver-
sion of cellulose into platform chemical compounds. Tan and Lee [136]
had presented a novel and environmental friendly pre-treatment pro-
cess that could enhance enzymatic hydrolysis of macroalgae cellulosic
residue (MCR) to glucose using Dowex (TM) Dr-G8 as catalyst. The op-
timum yield of glucose reached 99.8% under the optimal condition for
solid acid pre-treatment (10% w/v biomass loading, 4% w/v catalyst
loading, 30 min, 120 °C) followed by enzymatic hydrolysis
(45 FPU·g−1 of cellulase, 52 CBU·g−1 of β-glucosidase, 50 °C, pH 4.8,
30 h). [136]
3. Bioethanol Conversion Technologies
Bioethanol is produced from macroalgae through fermentation pro-
cess by yeast or bacteria [141]. Various macroalgae species have differ-
ent chemical and physical characteristics which required specific
process parameters. Thus, it is important to understand its characteris-
tics prior to production of bioethanol. A complete assessment of the fer-
mentation process is generally referred to cell growth profile,
consumption of reducing sugar by microorganism and bioethanol pro-
duction rate [142]. There are numerous fermentation methods used to
convert reducing sugar produced from macroalgae into bioethanol.
The processes are denoted as follows: (1) separate hydrolysis and fer-
mentation (SHF); (2) simultaneous saccharification and fermentation
(SSF); (3) simultaneous saccharification and co-fermentation (SSCF);
and (4) consolidated bioprocessing (CBP) [112].
3.1. Separate hydrolysis and fermentation (SHF)
Separate hydrolysis and fermentation (SHF) process using chemicals
(in the acid hydrolysis and fermentation) or enzymes (in the enzymatic
hydrolysis and fermentation) is suitable to completely convert the re-
ducing sugars from biomass into bioethanol. In general, the saccharifica-
tion/hydrolysis process (dilute acid hydrolysis or enzymatic hydrolysis)
is carried out first in a separate vessel followed by fermentation of the
hydrolysate into bioethanol in a different vessel at different conditions.
Table 9 summarizes the bioethanol production from SHF tested on var-
ious types of macroalgae.
Most of researches had reported on the utilization of S. cerevisiae in
their fermentation process. Wu et al. [143] reported that the bioethanol
production from acid hydrolysed Gracilaria sp. resulted in 0.236 g of
bioethanol production from 1 g of dry macroalgae, or corresponding
to 81% theoretical yield. In addition, Meinita et al. [101] used
S. cerevisiae directly for bioethanol fermentation from K. alvarezii hydro-
lysate. The bioethanol yield achieved after 24 h fermentation at 30 °C
from 30.5 g·L−1 of reducing sugars was 0.21 g·g−1 galactose, which
corresponded to a 41% of theoretical yield. Hargreaves et al. [98] ex-
plored the usage of carrageenan hydrolysate from K. alvarezii and was
fermented using a galactose-strain of S. cerevisiae. An optimum
bioethanol concentration of 37 g·L−1 was attained after 12 h of fermen-
tation, corresponding to a 90.6% theoretical yield.
Borines et al. [33] reported that after fermentation by S. cerevisiae at
40 °C for 48 h, the bioethanol conversion rate of the enzyme hydrolysate
from Sargassum spp. reached 89%, which was markedly higher than the
theoretical yield of 51% based on glucose as substrate. Thus, it can be
concluded that the fermentation of glucose or galactose after hydrolysis
can be fermented by the yeast of S. cerevisiae.
3.2. Simultaneous saccharification and fermentation (SSF)
Simultaneous saccharification and fermentation (SSF), which com-
bines enzymatic hydrolysis and fermentation into a single step, is an-
other alternative for the production of bioethanol from biomass. SSF is
usually preferred over the SHF process since inhibition of cellulase can
be minimized and high bioethanol production rate can be achieved
due to rapid conversion of glucose into bioethanol by yeast [145,146].
In addition, SSF process posed several advantages such as low risk of
contamination, small initial osmotic stress of fermenting agent, and
high energy efficiency [147]. Table 10 shows the bioethanol production
from SSF tested on various types of macroalgae.
Hargreaves et al. [98] evaluated the potential of cellulosic residue
from K. alvarezii as feedstock for bioethanol production and attained
53 g·L−1 of bioethanol via SSF process. In addition, bioethanol produced
from pretreated S. japonica via SSF process using different fermentation
mediums such as sodium citrate buffer, deionized water, and liquid hy-
drolysate exhibited insignificant differences. Nevertheless, deionized
water is found to be cost-effective and highly efficient to produce
bioethanol from the treated macroalgae. In SSF process, a bioethanol
concentration of 6.65 g·L−1 was obtained, which corresponding to a
 I.S. Tan et al. / Chinese Journal of Chemical Engineering 28 (2020) 502–517 513

Table 9
Bioethanol production from SHF tested on red and brown macroalgae

Macroalgae feedstock Conditions Fermentation process conditions Theoretical bioethanol yield/% References

Red macroalgae
Gracilaria sp. H2SO4 + enzyme 10% v/v S. cerevisiae Wu Y2, 30 °C, pH 4.5, static for 48 h 81 [143]
(whole biomass)
Gracilaria verrucosa Enzyme 6% v/v S. cerevisiae HAU, 30 °C, pH 6 for 16 h 84.31 [111]
(pulp after agar extraction)
Kappaphycus alvarezii (carrageenan) H2SO4 10% v/v S. cerevisiae, 28–30 °C for 168 h 4.6 [144]
Kappaphycus alvarezii H2SO4 5% v/v S. cerevisiae NCIM, 30 °C, pH 6.4–6.8, 150 r·min-1 for 48 h 82 [103]
(Granule)
Kappaphycus alvarezii H2SO4 S. cerevisiae, 30 °C, pH 5, 120 r·min-1 for 72 h 41 [102]
(whole biomass)
Kappaphycus alvarezii (carrageenan) H2SO4 7 g·L−1 S. cerevisiae CBS1782, 30 °C, pH 5, 150 r·min-1 for 24 h 90.6 [98]

Brown macroalgae
Laminaria japonica H2SO4 + enzyme 0.375 g·L−1 S. cerevisiae, 30 °C, pH 6.5 for 36 h 80.8 [108]
(floating residue)
−1
Sargassum spp. H2SO4 + enzyme 0.5 g·L S. cerevisiae, 40 °C, pH 4.5 for 48 h 89 [33]
(whole biomass)
−1 -1
Ulva fasciata Hot buffer + enzyme 109 CFU·mL S. cerevisiae, 28 °C, 120 r·min for 48 h 88.2 [74]

glucose equivalent concentration of 13.01 g·L−1, or SSF yield of 67.41%
based on the total available glucan of the pretreated S. japonica [149].
In addition, Kim et al. [148] reported that SSF was superior to SHF for
bioethanol production from G. amansii with bioethanol conversion yield
of 76.9% after 24 h. Adams et al. [85] evaluated the production of
bioethanol from Saccharina latissima macroalgae samples using
S. cerevisiae and 1 U·kg−1 laminarinase. They showed that the highest
bioethanol yield of 0.45% (v/v) was successfully achieved [85]. More re-
cently, Korzen et al. [14] used Ulva rigida (green seaweed) as a feedstock
to produce bioethanol through SSF process assisted with sonication. A
maximum of 6.2% of bioethanol (on dry wt. basis) was obtained under
sonication in 3 h vs. only 4.9% bioethanol under incubation even after
48 h.
3.3. Simultaneous saccharification and co-fermentation (SSCF)
In contrast to lignocellulosic biomass, macroalgae contains cellulose
and galactan/carrageenan that can be converted to glucose and galac-
tose, respectively, after hydrolysis process. In order to achieve more eco-
nomical route of macroalgae-based biorefinery, the microorganisms in
fermentation process should be able to consume all reducing sugars si-
multaneously and efficiently [150]. However, carbon catabolite repres-
sion (CCR) in microorganisms helps to obtain a proper balance
between their metabolic capacity and the maximum sugar uptake capa-
bility. Generally, glucose is the preferred carbon source and the utiliza-
tion of other carbon sources may be inhibited until the glucose content
is depleted [150]. Thus, sequential utilization or diauxic growth was ob-
served during mixed sugar fermentation, resulting to low yield and pro-
ductivity of bioethanol [151].
In order to overcome these problems, researchers had recently dem-
onstrated SSCF of cellulose and galactose [98]. The galactose liquid frac-
tion was mixed with the cellulosic residue in a concentration of 18% (w/
v) prior to enzymatic hydrolysis for 24 h. Thereafter, S. cerevisiae
CBS1782 (was initially screened for its ability to ferment galactose)
was introduced to the medium for bioethanol production. The result
showed a significant improvement in the consumption of galactose dur-
ing the fermentation process.
In another study, Park et al. reported the usage of a novel
ethanogenic strain S. cerevisiae ATCC2341 that was able to utilize galac-
tose for fermentation [152]. This mutant yeast strain exhibited excep-
tional fermentative performance on galactose and a mixture of
galactose and glucose. At 120 g·L−1 of initial galactose concentration,
bioethanol concentration reached 6.9% (v/v) within 36 h with 88.3% of
theoretical bioethanol yield. The results obtained herein demonstrated
a great opportunity for bioethanol production from macroalgae, which
mainly consisting of galactose.
3.4. Direct microbial conversion (DMC) or consolidated bioprocessing
(CBP)
Direct microbial conversion (DMC) or consolidated bioprocessing
(CBP) is a process in which all enzymes and bioethanol are produced
in a single reactor by a single microorganism community [153,154].
The key difference between DMC and other fermentation process is
that only one microbial community carries out both production of cellu-
lase and fermentation [152]. Wargcki et al. [156] had shown that
bioethanol could be produced directly from engineered alginate lyases
from Vibrio splendidus and functional alginate transport into Escherichia
coli. Through this method, 0.281wt%. of bioethanol yield was attained
which exceeded ideal bioethanol yield of 0.245wt%. for macroalgae.
More recently, Trivedi et al. [157] reported the isolation of cellulase
from marine fungus Cladosporium sphaerospermum through DMC pro-
cess. The isolated cellulose was applied in saccharification of
U. fasciata (green macroalgae) with attained bioethanol yield of

Table 10
Bioethanol production from SSF tested on various macroalgae

Macroalgae feedstock Fermentation process conditions Theoretical bioethanol References

yield/%

Red macroalgae
Kappaphycus alvarezii (cellulosic residue) 7 g·L−1 S. cerevisiae CBS1782, 18% w/w biomass loading, 30 °C, pH 5, 150 r·min-1 for 24 h 78.5 [98]
Gelidium amansii 5 mg S. cerevisiae KCTC7906, 15% w/v biomass loading, 37 °C, pH 4.8 for 24 h 76.9 [148]

Brown macroalgae
Saccharina latissima 0.5% (v/v) S. cerevisiae, 32 °C, pH 6 for 48 h – [85]
Saccharina japonica 10% v/v S. cerevisiae DK 410362, 6% w/v biomass loading, 43 °C, pH 5, 130 r·min-1 for 48 h 67.41 [149]

Green macroalgae
Ulva rigida 1.25% w/v S. cerevisiae, 4.2% w/v biomass loading, 37 °C, pH 5, 150 r·min-1 for 48 h 66.6 [14]
514 I.S. Tan et al. / Chinese Journal of Chemical Engineering 28 (2020) 502–517
0.47 g·g−1 reducing sugar, which corresponded to 93.8`% conversion
efficiency.
3.5. Cost estimation of bioethanol production
Cost estimation of bioethanol production had been reported by
many researchers in order to evaluate the economic feasibility of
bioethanol production [158-160]. Nevertheless, there was still a lack
of research being carried out to clearly define the bioethanol production
cost from macroalgae, as well as price competitiveness compared with
gasoline. In fact, studies on economics of industrial-scale production of
bioethanol from macroalgae are still very rare in literature. Economic
analysis has been used as a promising tool to assist the biofuels research
community in identifying key cost drivers, evaluating novel technolo-
gies and assessing new process configurations [161]. Rigorous economic
analysis requires a sound technical understanding of the process and es-
tablishing mass and energy balance estimates on a process-unit-level
basis. This is an important precedent to develop associated unit opera-
tion capital and operating cost estimates that can seamlessly be inte-
grated into a production pathway assessment.
According to Philippsen et al. [162], a general well-to-wheel model
of macroalgae-based bioethanol production was developed and applied
to the case of bioethanol produced from S. latissima, which was farmed
in British Columbia, Canada. This study showed that, despite the chal-
lenges of high-water content, high ash content, and limited harvest sea-
sons, macroalgae-based bioethanol was a promising biofuel with low
Carbon Intensity (CI), high Energy Return on energy Invested (EROI)
and good potential for commercialization. Reith et al. [63,163] had
used 100000–500000 ton of dry laminaria biomass per year to estimate
the preliminary cost for bioethanol production. Through this study, the
maximum seaweed price (MSP) of $ 28/dry ton was determined for a
500000 t·a-1 plant [63,163]. However, it is also expected that the actual
macroalgae bioethanol price is likely to be higher since the calculations
were based on optimistic conditions and assumptions.
4. Life Cycle Assessment (LCA) of Macroalgae Biofuels
Up to now, only a few number of LCA studies had been carried out
with respect to the potential of macroalgae biomass as a raw material
in the production of biogas and bioethanol [164-168]. Aitken et al.
[165] performed an analysis of the life cycle assessment of biofuel pro-
duction from offshore grown brown macroalgae (Gracilaria chilensis
and Macrocystis pyrifera), focusing on different offshore cultivation
methods (bottom planting and long-line cultivation) in Nordic condi-
tions. The study suggested that under base case conditions, bottom
planted G. chilensis was the most suitable option to produce both
bioethanol and biogas due to the low input method of cultivation. In ad-
dition, by using advanced cultivation and processing methods of long-
line cultivated M. pyrifera could result to a more sustainable biofuels
production. If these methods could be implemented at a commercial
scale, the production of biofuel from macroalgae could achieve long
term sustainability compared to other biomass feedstocks.
Alvarado-Morales et al. [164] studied bioenergy production from
offshore-cultivated brown macroalgae (Laminaria digitata) in Nordic
conditions based on two production scenarios: one was producing bio-
gas only and the other was producing bioethanol with stillage to be con-
verted to biogas. The production of both bioethanol and biogas
generated a higher energy export compared to with solely biogas. This
was preferable in terms of its energy ratio. Nevertheless, the additional
fermentation and distillation process resulted in poorer environmental
impacts. This study also suggested that macroalgae biomass was an al-
ternative and promising feedstock for bioethanol production.
In addition, the LCA study on utilization of macroalgae to enhance
CO2 fixation and biofuels production was performed by Aresta et al.
From the study, it was demonstrated that there was potential energy
which could be beneficial when associated with fixation of CO2 by
macroalgae, especially when effluent water was used as nutrients
source to cultivate macroalgae. Nevertheless, the net energy gain was
depended on the conversion technology. In the best case considered
so far, macroalgae could generate a net energy of 11000 MJ·t−1 dry
algae compared to 9500 MJ·t−1 relevant to microalgae gasification
[167].
Nevertheless, the life cycle assessments on macroalgae biofuel pro-
duction are still limited in the literature and it is still challenging to
scale-up the macroalgae bioethanol production based on current tech-
nology maturity. The technologies selected for bioethanol production
should be further verified not only from the standpoint of economic po-
tential, technical feasibility, but also from the environmental
perspective.
5. Conclusions
In view of macroalgae biomass as a potential source of clean, sustain-
able and renewable biofuel worldwide, there is intensified research
targeting macroalgae-based bioethanol technology with an intention
to compete with or even replace the fossil fuels. The usage of
macroalgae biomass for bioethanol production is still under develop-
ment and most of the literatures are still limited to lab-scale study. In
order to open new possibilities for cost-effective industrial application,
future research could focus on technology development for genetic en-
gineering, where the carbohydrates content can be significantly in-
creased. In addition, hydrolysis process with high substrate loading
and efficient microbial utilization of reducing sugars are required.
At the current stage, macroalgae biomass is an attractive renewable
source for third-generation bioethanol production since it possesses in-
herent advantages over previous generations. However, the high cost of
conversion is the main stumbling block that may take longer time for
the macroalgae bioethanol to triumph the market than previously
projected. Therefore, improving the conversion process efficiency into
bioethanol will be vital to establish this emerging source of bioenergy
for commercial utilization. If these technologies are further optimized,
the production of bioethanol from macroalgae could be one of the viable
processes in the near future.
Acknowledgements
The authors would like to acknowledge the financial support re-
ceived from the Ministry of Higher Education (MOHE) Malaysia through
Fundamental Research Grant Scheme Malaysia's Rising Star Awards
2016 (FRGS MRSA 2016) with cost centre 203/PJKIMIA/607136. Sup-
port from Ministry of Education Malaysia through HiCOE award to
CBBR is duly acknowledged.
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