Renewable and Sustainable Energy Reviews 111 (2019) 44–56

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Renewable and Sustainable Energy Reviews

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Potentials and challenges in lignocellulosic biofuel production technology T

a,∗ a b b
M. Raud , T. Kikas , O. Sippula , N.J. Shurpali
a
Institute of Technology, Estonian University of Life Sciences, Kreutzwaldi 1, Tartu, 51014, Estonia
b
Department of Environmental and Biological Sciences, University of Eastern Finland, P.O. Box 1627, Kuopio, FI-70211, Finland

ARTICLE INFO ABSTRACT

Keywords: Since the Kyoto protocol, the EU renewable energy policy has been the driving force in research and develop-
Lignocellulose ment of the production and use of biofuels. Therefore, the utilization of biofuels and the scientific research to
Biofuel widen the scope of their commercialization are being increasingly promoted. In this context, we present here an
Biomass overview of the technological developments that have occurred in the production of different biofuels and their
Bioethanol
resources with a focus on the lignocellulosic biomass and biofuels derived from it. In the recent years, many
Thermochemical process
technologies have evolved enabling the production of different liquid biofuels from lignocellulosic biomass.
Bioethanol production using microbial fermentation, and production of bio-oil using fast pyrolysis process of
biomass are some of the most widely researched and promising technologies. The first production plants based
on these techniques are already operational. Bioethanol production from lignocellulosic biomass promises a
good fuel yield under laboratory scale. However, there are several challenges that need to be tackled before the
production process can be commercialized. We provide here an overview of the various possibilities that can be
exploited for biofuel production and make recommendations for increasing the production efficiency with a view
to improving the overall yield and lowering the production costs.

1. Introduction – importance and benefits of liquid biofuels
Climate change is a global problem. Every nation needs to partici-
pate in finding solutions to this problem with active, shared measures.
The carbon dioxide content of the atmosphere has increased by 43%
since 1750 [1]. The atmospheric CO2 concentration as measured at the
Mauna Loa laboratory during March 2019 was 413 ppm (as opposed to
about 280 ppm during the preindustrial times). The growth rate of at-
mospheric CO2 during the 1960-69 decade was 0.85 ppm yr−1, while it
climbed to 2.28 ppm yr−1 during the recent 10-year (2008–17) period.
The reason for such a drastic change in the atmospheric composition is
attributed to our overdependence on fossil fuels for energy and defor-
estation. Corresponding to this rise in CO2 content, the mean global
surface temperature has already increased by 0.85 °C, compared to the
preindustrial era [2]. The impacts of a changing climate are becoming
evident across all continents in the warmer oceans, reduced snow and
ice cover and rising sea levels.
Today, fossil fuels account for nearly 80% of the primary energy
consumed in the world, of which 58% alone is consumed by the
transport sector [3,4]. As the global demand for fossil fuels is increasing
and their supply is dwindling, there is a need for alternative fuels that
are renewable and sustainable. Biofuels are an important alternative
with a potential to replace traditional fossil fuels [5]. Biofuels have
emerged as one of the most strategically important sustainable fuel
sources and are considered important in our progress towards limiting
greenhouse gas emissions, improving the environmental health and
finding new energy resources.
Biomass and waste, as a global energy source, account for over 70%
of all renewable energy production. Its contribution to final energy
consumption in 2015 was approximately on par with that of coal. Direct
burning is the largest end use of biomass and waste [6]. However,
various liquid biofuels can be produced from such feedstocks using
different thermochemical or biochemical processes and technologies
[7]. The technologies for producing ethanol from sugar and starch-
based feedstocks, and biodiesel from vegetable oils and other lipid
feedstocks are well-established, and provide most of today's transport
biofuels. However, the sustainability of biofuels produced in competi-
tion with food crops is questionable because of the conflict with food
supply [3,8].
Since the demand for biofuels and food has been increasing, a lot of
attention in recent years has been directed at the utilization of lig-
nocellulosic biomass [9,10] derived from either non-edible residues of
food crops or non-edible whole plant biomass. Different biomass types
such as by-products (cereal straw, sugar cane bagasse, forest residues),

∗
Corresponding author.
E-mail address: Merlin.raud@emu.ee (M. Raud).

https://doi.org/10.1016/j.rser.2019.05.020
Received 3 July 2018; Received in revised form 3 May 2019; Accepted 8 May 2019
Available online 15 May 2019
1364-0321/ © 2019 Elsevier Ltd. All rights reserved.
M. Raud, et al.
Abbreviations
AFEX Ammonia fiber explosion
CBP Consolidated bioprocessing
EU European Union
FPBO fast pyrolysis bio-oil
GHG Greenhouse gas
wastes (organic components of municipal solid wastes), and dedicated
feedstocks (forage grasses, short rotation coppices and other energy
crops) can be used for producing biofuels [3,4]. The main advantage of
the production of biofuels from non-edible feedstocks is that it limits
the direct food versus fuel competition [8,11,12].
Different liquid fuels can be made from lignocellulosic feedstock
using either biochemical or thermochemical route [12,13]. Most
common approach is to make ethanol or butanol using biochemical
route and other synthetic fuels using thermochemical route. However,
the processing technologies of these biofuels are immature yet. Hence,
significant progress needs to be made in terms of perfecting the pro-
duction process [14]. Of these technologies, lignocellulosic ethanol
production through the biochemical route and fast pyrolysis bio-oil
production via thermochemical route have been scaled up to the early
market development phase, while other thermochemical technology is
still in demonstration stage and other biochemical routes are mainly
still operational at the lab-scale [6]. In view of the above, the goal of
this paper is to provide an overview of the state-of-the-art in liquid
biofuel production technologies from lignocellulosic biomass.
2. Government initiatives and EU legislation for biofuels
The aim of the Kyoto Protocol adopted in 1998 was to reduce
greenhouse gas emissions during 2008-2012 by 5%, compared to 1990.
Under this protocol, the EU had the obligation to reduce greenhouse gas
emissions by 8%. This responsibility was shared among the Member
States so that the overall reduction would be 8% [15]. In order to
achieve this target, the EU Member States set a goal to significantly
increase the use of renewable energy sources in all fields including
transportation [16] and a series of measures were introduced to im-
prove the development of energy from renewable sources.
The European Parliament and EU Council adopted the biofuel di-
rective in 2003 and defined different types of energy that could po-
tentially be generated from various forms of renewable sources. With
this directive, a target was set for the first time on the proportion of
biofuels and other renewable fuels to be mixed with petrol and diesel
fuels used for transport purposes. A reference value was set so that
Fig. 1. Classification of biofuels according to processing technology and biomass type.
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Renewable and Sustainable Energy Reviews 111 (2019) 44–56
IL Ionic liquids
LHW Liquid hot water
NED N2 explosive decompression
SE Steam explosion
SHF Separate hydrolysis and fermentation
SSCF Simultaneous hydrolysis and co-fermentation
SSF Simultaneous hydrolysis and fermentation
biofuels and other renewable fuels are used to the extent of at least
5.75% by 31 December 2010 in the EU member states. This reference
value was calculated on the basis of the fuel energy content. Also, an
intermediate target of 2% was set by December 2005 [17]. However,
the share of biofuels in the transport sector was only 0.9% in 2005 and
3.8% in 2010 [18].
A Renewable Energy directive approved in 2009 set targets for all
EU countries to make renewable energy sources account for 20% of EU
energy, 20% GHG emission reduction and 20% of energy efficiency
increase by 2020. Each member state was assigned specific national
binding targets. A 10% target of energy from renewable resources in the
transport sector was also set to all member states. In order to achieve
these targets each EU state was obligated to prepare a national re-
newable energy action plan for 2020 and to set support schemes in
order to encourage and promote energy efficiency and utilization of
renewable resources [19]. Since the approval of these directives, the
consumption of renewable sources has increased and in 2014, renew-
able energy sources accounted for a 12.5% share of the EU-28's gross
inland energy consumption. At the same time, the average share of
energy from renewable sources in transport increased from 1.4% in
2004 to 6.7% in 2015 [20].
In 2012, the European Commission stated in its proposal that only
advanced biofuels, with low estimated indirect land use, climate change
impacts, and high overall greenhouse gas savings should be supported
as part of the post 2020 renewable energy policy framework [21].
According to the European alternative fuel strategy, adopted in 2013,
increasing demand for energy [21]and reduced dependency on oil in
transport sector could be met only by a comprehensive mix of alter-
native transport fuels. Therefore, due to the concerns over food security
and land use change, it was proposed to set a limit of 5% on the amount
of first generation biofuels that can be counted towards the Renewable
Energy Directive targets. The aim was to increase the incentives for
advanced biofuels made from lignocellulosic biomass, residues, waste,
and other non-food biomass, including algae and microorganisms [22].
These suggestions were taken into account in the directive (EU)
2015/1513 that repeats the goal of 20% energy efficiency by 2020 set
in the previous directive. In addition, different types of new energy
M. Raud, et al.
substrates such as waste, starch-rich crop, lignocellulosic material,
processing residues etc. were defined and the transition from conven-
tional first-generation biofuels to advanced second-generation biofuels
was aimed. Therefore, a maximum limit of 7% of conventional first
generation biofuels in transport sector by 2020 was introduced [23].
The remaining 3% must not be based on food crops, but may come from
a variety of multiple counted alternatives as well as biofuels from used
cooking oil, animal fats and from advanced biofuels like second-gen-
eration biofuels [16].
Energy Roadmap 2050, which aims to decarbonize the EU economy
by 2050, states that the renewables have the potential to provide ap-
proximately 30% of total EU energy consumption by 2030 [24]. In
order to gain these goals, concrete milestones must be set. Therefore, in
2014 a new greenhouse gas reduction target of 40% and a share of
renewable energy in the EU of at least 27% by 2030 was agreed. In
addition, it was also proposed that it is not appropriate to establish new
targets for fuels used in the transport sector after 2020 [25]. This target
is binding at EU level and would be fulfilled through individual Member
State contributions. With a proposal of new and revised Renewable
Energy Directive published in 2016 a goal was set to make the EU a
global leader in renewable energy and to ensure that the target of at
least 27% renewables in the final energy consumption in the EU by
2030 is met. In addition, a limit of 7% of conventional first generation
biofuels in transport sector should be reduced to 3.8% by 2030 [26].
3. Classification of biofuels
Different classification systems for biofuels have been proposed. The
most widely used classification of biofuels according to processing
technology and biomass type has been outlined in Fig. 1. Firstly, bio-
fuels can be classified as primary and secondary biofuels according to
their production process. Primary fuels are produced from biomass in
its natural form such as fuelwood, wood chips and pellets etc. Primary
fuels are mainly used for cooking, heating and electricity production.
Secondary biofuels are gained via processing of biomass. List of sec-
ondary biofuels include charcoal, ethanol, diesel, biogas, etc. These
fuels can be used in a variety of applications such as transport, energy
production or various industrial applications [8]. The secondary fuels
can be additionally grouped according to biomass type and processing
technology into different fuel generations [27].
The first generation biofuels, conventional biofuels, are produced
primarily from food crops such as grains, sugar cane and oil cultures
etc. [7,28,29] by using simple production technology [8]. The most
widespread first generation fuel is ethanol, produced from sugar or
starch-rich biomass by fermentation with yeast, which converts the
sugars to ethanol. Additionally, hydrolysis is used before fermentation
when the starch containing grains are used. Another first generation
biofuel is biodiesel from vegetable oils. Biodiesel is usually produced by
the transesterification process or cracking [8]. The first generation
biofuel production technologies have been successfully established on
firm foundations and are widely used, as their technologies and im-
plementation are simple [29]. However, the viability of first-generation
biofuels is questionable because of the conflict with food supply
[3,8,30].
Second generation biofuels are called lignocellulosic biofuels since
they are produced from non-edible biomass such as energy crops,
agricultural and forest residues, which are cheap, abundant and re-
newable [7,28,29]. The main advantage of the production of second-
generation biofuels is that it limits the direct food versus fuel compe-
tition issues associated with first generation biofuels [8,11,12], as in-
edible biomass is used. Second-generation biofuels can be produced
using two distinctive ways – thermochemical and biochemical route.
Thermochemical processing involves heating the biomass in the ab-
sence or in the presence of different concentrations of oxygen. Thus, by
thermal decay and chemical reformation, wide range of products can be
produced. Biological conversion technologies, on the other hand, are
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Renewable and Sustainable Energy Reviews 111 (2019) 44–56
based on microbial and enzymatic processes for producing sugars from
biomass [12,14] and then converting the sugars to fuels such as ethanol
or butanol. It is worth noting here that compared to first generation
bioethanol, currently the second generation biofuels have lower effi-
ciency due to a lower yield of the final product [30].
The third generation biofuels are produced using algae or other
aquatic biomass and fourth generation biofuels are produced using
genetically modified or engineered algae and microorganisms that are
able to capture CO2 from the atmosphere [7,29]. Algal biomass is a
promising alternative feedstock due to its high lipid and carbohydrate
contents, ability to be easily cultivated in water environment and re-
latively low land usage with high atmospheric CO2 uptake capabilities
[31]. Therefore, algae are a potential source of different fuels such as
biogas via anaerobic digestion or biodiesel produced from lipids accu-
mulated in the algae cells [32]. However, the industrial cultivation and
production technologies of algal and fourth generation biofuels are still
in infancy and the production process is currently expensive and un-
profitable.
4. Biofuel resources – lignocellulosic biomass
Biomass burning (harvested organic matter) has been fulfilling hu-
mankind's energy needs for millennia. While such fuels remain the
primary energy source for many people in developing and emerging
economies, such traditional use of biomass is not sustainable. Moreover,
inefficient combustion leads to harmful emissions with serious health
implications. There exist modern technologies that can convert this
organic matter to solid, liquid and gaseous forms that can more effi-
ciently provide for energy needs and replace fossil fuels [12]. Lig-
nocellulosic biomass is one of the most abundant natural resources and
therefore, has become the major focus of research and development as a
promising renewable resource for the production of different biofuels
[33]. Lignocellulosic materials are particularly attractive as feedstocks
for biofuel production because of their relatively low cost, great
abundance, and sustainable supply [30].
A wide range of lignocellulosic biomass feedstocks can be used as
sources of bioenergy. They include residues and co-products from
agricultural and timber industry [34], crops grown for energy [35],
food crops (corn, wheat, sugar and other raw materials [36]), non-food
crops, such as perennial lignocellulosic plants (e.g. grasses, such as
Miscanthus [37], reed canary grass [38,39] and trees, such as short-
rotation willow [40] and eucalyptus [41] and oil-bearing plants (ja-
tropha and camelina [42]). Traditional lignocellulosic feedstocks in-
clude mainly different energy crops and agricultural residues [43],
which are widespread in all regions. Agricultural residues include a
variety of agricultural wastes such as corn stover, cornstalks, rice and
wheat straws as well as sugarcane bagasse [44]. On the other hand,
dedicated energy crops are purposely grown and harvested as biomass
for energy use. Dedicated energy crops have a low input in require-
ments, high biomass yield, high cellulose content, easy to grow (with
broad cultivation in a short period of time) and tolerance to drought,
high temperatures, flooding and salt stress [3]. Moreover, this kind of
biomass can be produced using less farmland, since the whole plant can
be utilized as raw material, while first generation uses only seeds [7]. In
addition, forest residues, wood process wastes, industrial residues and
urban greening wastes can be used as lignocellulosic feedstocks
[4,44,45].
The potential lignocellulosic material from forestry in central
Europe and Scandinavian countries are aspen, alder, birch, willow and
poplar. Agricultural residues such as crop-derived wastes include re-
sidues of grains, corn, flowers, grass, and straw [46]. Currently the most
promising and abundant cellulosic feedstocks, derived from plant re-
sidues in the U.S., South America, Asia and Europe, are from corn
stover, sugarcane bagasse, rice and wheat straws, respectively. Amongst
dedicated energy crops, switchgrass (Panicum virgatum) is the primary
herbaceous prairie grass and energy crop grown in the North America
M. Raud, et al.
namely, Canada and the U.S while Miscanthus is a suitable biomass
candidate for biofuel production in Asia and Europe [47].
4.1. Biomass structure and components
The lignocellulosic biomass is mainly composed of plant cell walls,
which provide strength and resilience and form the woody tissue of the
plants [3]. Cell wall consists mainly of cellulose, hemicellulose, and
lignin, which are organized into a complex structure as seen in Fig. 2.
The typical percentages of dry weight are 35–50% cellulose, 20–35%
hemicellulose, and 5–30% lignin, depending on the biomass type,
source and even on harvesting time [30,48,49]. Generally, the crop
residues contain more hemicellulose and woody residues have higher
lignin content. The composition of different types of biomasses are
shown in Table 1. These major constituents of biomass exist as a closely
associated, rigid network in plants [33], which makes it recalcitrant to
decomposition by enzymes and microbes [33,50].
Cellulose (C6H10O5)x, the main constituent of lignocellulosic bio-
mass, is a polysaccharide that consists of a linear chain of D-glucose
monomers, linked together by β-(1,4)-glycosidic bonds [58]. The cel-
lulose chain consists of 10 000 monomers in wood and up to 15 000
monomers in cotton [30]. The linear cellulose chains are associated
together via intra- and intermolecular hydrogen bonds to make long
tightly packed cellulose fibers [3,30]. These cellulose microfibers, or-
dered into tightly packed and highly crystalline structures, are insoluble
in water and highly resistant to depolymerisation [58]. There are also
some parts of cellulose that are less crystalline - amorphous regions.
These areas are less resistant to biodegradation and thus, enzymes are
able to bind to cellulose there and start hydrolyzing process [30]. It has
been shown that the most important property of biomass is high cel-
lulose content since bioethanol is produced from glucose formed during
cellulose hydrolysis while low lignin content is preferred since the de-
gradation products of lignin and hemicellulose inhibit the hydrolysis of
cellulose to sugars [59–63].
Hemicellulose is the second most abundant polymer in biomass.
Unlike cellulose, hemicellulose is not chemically homogeneous [30].
Hemicelluloses (C5H8O4)m are a group of polysaccharides located in
secondary cell walls, which interact with cellulose and lignin. They are
heterogeneous branched amorphous biopolymers of pentoses (xylose,
arabinose), hexoses (mannose, glucose, galactose) and acetylated
Fig. 2. The structure of lignocellulosic biomass.
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Renewable and Sustainable Energy Reviews 111 (2019) 44–56
sugars [3,50,58]. Due to their low molecular weight and branched
structure with short lateral chains, the hemicelluloses are easily hy-
drolyzed [3,30,50].
Hemicelluloses bind to cellulose microfibers via hydrogen-bonds by
covering them and thus form a network that provides the structural
backbone to plant cell wall [58]. This hemicellulose cover makes the
cellulose fibers difficult to access for enzymes, which are needed to
convert the biomass into fuels. It has been proposed that in order to
significantly increase the cellulose digestibility and conversion effi-
ciency, at least 50% of the hemicellulose should be removed with
pretreatment [30]. During processing, the severity parameters like
temperature and retention time must be controlled to avoid formation
of hemicellulose degradation products such as furfurals and hydro-
xymethyl furfurals, which later inhibit the fermentation process
[30,50].
Lignin [C9H10O3(OCH3)0.9–1.7]n is an aromatic amorphous hetero-
polymer synthesized from phenyl propanoid units, primarily of syr-
ingyl, guaiacyl and p-hydroxy phenol, which are held together by dif-
ferent linkages to make a complicated matrix [50]. Lignin forms a
three-dimensional network in which cellulose and hemicellulose fibers
are embedded [48] and therefore, lignin is generally accepted as the
‘glue’ that binds the different components of lignocellulosic biomass
together [30]. This structure provides structural support, mechanical
reinforcement, waterproofing and barrier to enzymatic digestion for
biomass [3]. Compared to cellulose and other structural poly-
saccharides, lignin is more resistant to biological attack [64].
5. Liquid biofuels from lignocellulose and their production
technologies
The feedstock of second-generation liquid biofuels is mainly sourced
from the lignocellulosic residues of agricultural and forestry sectors and
the dedicated energy crops [65]. Many processes are available to con-
vert lignocellulosic biomass into a product that can be used for elec-
tricity, heat or transport. Each of these bioenergy pathways consists of
several steps. These include biomass production, collection or har-
vesting, processing to improve the physical characteristics of the fuel,
pre-treatment to alter chemical properties, and finally conversion of the
biomass to useful energy. Conversion methods employed depend on the
type, location and source of biomass, and the technology used to
M. Raud, et al. Renewable and Sustainable Energy Reviews 111 (2019) 44–56

Table 1
Chemical constituents (%) of different lignocellulosic biomass types.
Cellulose Hemicellulose Lignin Other (ash, protein, extractives, etc.) Reference

Wheat straw 37–41 7 27–32 13–15 17–21 ± 2 [51,52]

Barley straw 33.8–37.5 21.9–24.7 13.8–15.5 [50,53]
Corn Stover 38,3 25,8 17,4 [50]
Sugar cane bagasse 40–43,1 27–37.5 10–20 [50,53]
Rice straw 31,1 22,3 13,3 [50]
Rice husks 38,4 16,1 16,7 26,1 [54]
Grasses 25–40 25–50 10–30 – [51]
Switch grass 39,5 25,0 17,8 – [50]
Miscanthus 38,2 27,1 26,4 [13]
Hardwood 45 ± 2 30 ± 5 20 ± 4 5,6 ± 3,2 [51]
Softwood 42 ± 2 27 ± 2 28 ± 3 3.5 ± 2.1 [51]
Forest residues 46,3 28,3 26,5 [55]
Poplar 43,8 14,8 29,1 [50]
Spruce 43,8 6,3 28,3 [50]
Garden biomass 39,75 26,55 24,46 [56]
Chemical pulp 60–80 20–30 2–10 [57]
Newspaper 40–55 25–40 2–30 [51,57]

provide the relevant final energy use [66,67].
For the production of liquid biofuels two fundamentally different
approaches are used - thermochemical or biochemical conversion
[14,47]. In case of thermochemical conversion, the biomass is heated
either in the absence of oxygen (pyrolysis) or in the presence of oxygen
(gasification) at high temperature, up to 800°. In pyrolysis, the ther-
mochemical conversion in the absence of oxygen converts the solid
feedstock to char, condensable gases forming so-called pyrolysis oil and
solid char residues. In gasification, the thermochemical process con-
verts the biomass into synthesis gas (syngas) such as hydrogen, CO, H2
and CO2 [47]. From syngas, a wide range of long carbon chain biofuels,
such as synthetic diesel, aviation fuel, or ethanol, can be reformed,
based on the Fischer–Tropsch conversion [68].
In the biochemical route however, the polysaccharides in the bio-
mass are converted into sugars using chemical or enzymatic hydrolysis.
Different sugars derived from lignocellulosic biomass can be further
used for production of different alcohols, lipids and biogas using spe-
cific microbial fermentation processes in production [65]. One of the
most common ways to produce biofuels from lignocellulose is to con-
vert the biomass into sugars and ferment the sugars into fuel. However,
due to the tight packaging of cellulose in lignin and hemicellulose
cover, the cellulose is difficult to access for cellulases. In order to in-
crease the process efficiency, it is necessary to break down lignin and
hemicellulose seal to make cellulose more accessible for further pro-
cessing [50]. The efficiency of cellulose hydrolysis is less than 20%
when pretreatment is not carried out. However, after pretreatment, the
efficiency can exceed 90% of the theoretical limit [69]. Therefore, to
make the conversion more efficient, biomass must be subjected to
pretreatment to open its structure and allow easier release of fermen-
table monosaccharides [70].
5.1. Bioethanol production technology
5.1.1. Pretreatment
Pretreatment is the crucial step for gaining high sugar yields from
polysaccharides. Over the years, many pretreatment methods have been
proposed and numerous review papers have been published, which give
an overview of the different methods and their mechanisms and
working principles [11,13,68,70–75]. Typical goals of the cost effective
pretreatment operation are to (1) produce highly digestible solids that
enable higher sugar yields during enzyme hydrolysis; (2) avoid the
degradation of sugar derived from cellulose and hemicellulose; (3)
avoiding the formation of inhibitors for subsequent hydrolysis and
fermentation steps; and (4) recovery of lignin [73]. The selection of
pretreatment method is made considering the delignification and
hemicellulose removal efficiency, low sugar losses, time savings,
economical issues, and environmental impact [76]. However, a single
pretreatment method is not always effective and therefore combination
of different pretreatment methods has been widely used. This way the
advantages of different pretreatment methods are combined and
thereby, improved overall efficiency of the process can be achieved.
Thus, the choice of a suitable method depends on the characteristics of
the biomass and the subsequent processing steps. According to working
principle and materials, the pretreatment methods can be divided into,
biological, physical, and chemical methods and additional methods that
combine characteristics of these [11,13,70–75]. The comparison of
different pretreatment methods is outlined in Table 2.
5.1.1.1. Biological methods. Biological methods are usually based on
different microorganisms that degrade the components of
lignocellulosic biomass. Brown, white and soft rot fungi have been
reported to produce enzymes that can depolymerize lignin and
hemicellulose in biomass [13,73,75] and thereby, enhancing
enzymatic hydrolysis of biomass [13]. Brown-rot fungi primarily
attack cellulose, whereas white and soft-rot attack lignin via the
production of extracellular enzymes called lignases [30,82]. Several
white-rot fungi, such as Ceriporiopsis subvermispora, Pleurotus ostreaus,
Ceriporia lacerata, Cyathus stercolerus, Pycnoporus cinnarbarinus and
Phanerochaete chrysosporium, have been examined and high
delignification efficiency with improved hydrolysis efficiency has
been reported [76,77]. The biological methods are environmentally
friendly and cheap to operate, since no chemicals are used. However,
the long process duration, due to low hydrolysis rate is the main
drawback of using fungi to pretreat woody materials [73]. A
combination of fungal pretreatment with other chemical/physical
pretreatment methods has shown an improvement in sugar and
biofuel yields [71].
5.1.1.2. Physical methods. Physical pretreatments include various
mechanical operations and different types of irradiation and
ultrasonic pretreatment [73]. Mechanical pretreatment enables to
achieve higher hydrolysis efficiency of biomass polymers and
improved digestibility. A combination of chipping, grinding and
milling are used to increase the surface area, reduce the chip size of
the biomass, and decrease the degree of polymerization and
crystallinity of cellulose [50,71]. The biomass chip size is usually
10–30 mm after chipping and 0.2–2 mm after milling or grinding
[50,51]. The pretreatments aimed for size reduction are very often
used in combination with other pretreatment methods as a first step in
the pretreatment process as a whole.
Extrusion pretreatment is a thermo-physical pretreatment in which
materials undergo mixing, heating and shearing, leading to physical

48
 M. Raud, et al.

Table 2
Comparison of different pretreatment methods.
Method Effect Advantages Disadvantages Reference

Biological
Microbial Degradation of lignocellulose components Low energy consumption, environmentally friendly, Low hydrolysis rate and efficiency [13,73,75]
no inhibitors
Fungal Degradation of lignin Low energy and chemical use, environmentally Low hydrolysis rate and efficiency [13,73,75–77]
friendly, no inhibitors
Chemical
Acid (sulfuric acid, hydrochloric acid, Hemicellulose dissolution, decreases cellulose High yield of sugars, high hemicellulose removal corrosion, formation of inhibitors, concentrated acids [50,68,71,72]
nitric acid) polymerization hydrolyze polymers to sugars, can be expensive due to
chemicals
Alkaline (sodium hydroxide, lime) Dissolution of lignin and a part of the hemicellulose, Operated at lower temperatures, does not require Residence time can be very long, formation of salts, can be [13,71,72,78]
decreases polymerization degree, increase of surface area complex reactors expensive due to chemical
Organosolv Fractioning and dissolution of biomass components High extraction efficiency, solvent recovery High cost, needs solvent recovery, flammability, volatility of [33,53,71]
especially lignin relatively easy solvents, low boiling point of solvents

49
Ionic liquids Selective dissolution of biomass components Highly effective, mild process conditions and High price of IL, IL can be toxic to enzymes, chemical [30,48,50,68]
chemicals, stable and non-volatile chemicals regeneration necessary
Physical
Comminuting (milling, grinding, Decrease particle size, degree of polymerization and No toxic compound formation High energy input [50,51,71]
extrusion, freezing) crystallinity
Irradiation (microwave) Lignin and hemicellulose removal Short process time and low energy demand, high not ready to be used on an industrial scale [50,56]
uniformity
Freezing Destruction of cell structure Cheap in Nordic areas Low efficiency [79]
Physio-chemical
Hot water Solubilisation of hemicellulose minimum inhibitor formation, uses lower High energy demand, involves large volumes of water [30,75]
temperatures, low cost of solvents
Steam explosion Destruction of biomass during decompression and Highly effective, chemical free Batch facilities do not permit processing large volumes [33,80]
dissolution of hemicellulose Incomplete degradation of lignin, the production of toxic by-
products
AFEX Reduces the lignin fraction Lignin removal, no/less inhibitor production and Inefficiency for biomass with high lignin content and the need [50,71,81]
surface area increase for high pressure, ammonia recycling is expensive
CO2 explosion Destruction of biomass during decompression and Almost no inhibitory components, uses lower High cost of equipment [30,71]
dissolution of hemicellulose temperatures, low cost of CO2
Renewable and Sustainable Energy Reviews 111 (2019) 44–56
M. Raud, et al.
and chemical alteration [50]. The treatment disrupts the lignocellulose
structure and causes the defibrillation, fibrillation and shortening of the
fibers and in the end, increases accessibility of carbohydrates to enzy-
matic attack [75]. Such physical pretreatment methods help avoid the
formation of inhibiting compounds [71], However, they are time-con-
suming, energy intensive, and expensive [51].
Other physical pretreatment methods, such as microwave exposure
and freezing, are usually used in combination with other more efficient
pretreatment methods [71]. Microwave irradiation alters the structure
of cellulose, degrades and partially removes lignin and hemicellulose
and increases the enzymatic susceptibility [50,56]. Novel method for
biomass is the freezing pretreatment, which changes the volume of
water during freezing. As the water impregnated biomass freezes, the
volume of water increases and this breaks the structure of the cell. This
method is suitable as an additional pretreatment method in areas where
natural cold period exists [79].
5.1.1.3. Chemical methods. In chemical pretreatment methods,
different chemicals such as acids, alkaline agents, organic solvents,
ionic liquids etc. are used to break down biomass components. The most
important characteristic of these methods is that they selectively and
efficiently isolate the different components of lignocellulose [72].
However, formation of toxic compounds, carbohydrate loss and high
cost of chemicals used in the process are common disadvantages [71].
Acid pretreatment with sulfuric acid is the most commonly used
chemical method for biomass pretreatment [83]. Acid hydrolyses the
hemicellulose into monosaccharides and this enables to gain better
access to cellulose for enzymatic hydrolysis. Acid-catalyzed processes
can be divided into two general approaches – concentrated acid used at
low temperature or dilute acid used at high temperature [50,68].
Concentrated acids (usually 72% sulfuric acid), used at low tempera-
ture, enable to gain high conversion rate of glucan in biomass to glucose
(up to 90%) [83]. However, the toxicity and corrosiveness of acids,
expensive acid recovery, and formation of fermentation inhibitors are
major drawbacks of this method [50]. Dilute acids (2–5% sulfuric
acids), on the other hand, are used at high temperature (120–200 °C)
and therefore, this approach demands more energy [83]. Dilute acid
process is industrially more attractive as lower amounts of fermentation
inhibitors are generated [50].
Alkaline pretreatments allow removal of significant amount of
lignin and thereby, highly digestible cellulose is gained [13,78]. Alka-
line pretreatment usually uses bases such as sodium, potassium, cal-
cium [50,83] to break the bonds between lignin and carbohydrates and
disrupt the lignin structure [53] and thereby, increase the enzymatic
accessibility of cellulose and hemicellulose [83]. The main benefit of
alkaline pretreatment is efficient lignin and hemicellulose removal as
well as its ability to increase the surface area for the reaction site for
further hydrolyses [71].
A new approach for biomass pretreatment uses different chemicals
as solvents. Organosolv pretreatment methods use different mixtures of
aqueous or organic solvents (such as ethanol, methanol, or acetone),
usually in combination with an acid or base catalyst [13,68]. The
treatment is used to break the internal lignin and hemicellulose bonds
[53], which leads to increased surface area and more accessible cellu-
lose fibers [33]. Depending on the operating parameters like tempera-
ture (180–195 °C), time (30–90 min), catalyst and liquid to biomass
ratio (4:1–10:1), the hemicellulose and lignin can be dissolved into
solution – black liquor and highly purified cellulose pulp can be gained
[53,72]. The method efficiently removes lignin and hemicellulose.
However, problems with the high cost of solvents and solvent recovery,
flammability and volatility, are its drawbacks [71].
Recently, much attention has paid to the use of ionic IL as solvents
for pretreatment of cellulosic biomass [30,75]. ILs are certain type of
organic salts composed of cations and anions, typically, large organic
cations and small inorganic anions [50]. ILs are a type of molten salts,
most of which have excellent solvency, low melting point, high thermal
50
Renewable and Sustainable Energy Reviews 111 (2019) 44–56
stability, non-volatility and designability [30,48,68,84]. The physical
and chemical properties of ILs can be adjusted to different reactions by
choosing different cations, anions and substituents [30,48,75,84]. ILs
are able to significantly change biomass structure by dissolving lig-
nocellulose components and breaking down bonds between cellulose,
hemicellulose and lignin in lignocellulosic materials [71]. ILs have the
superiority in dissolving carbohydrates and lignin simultaneously
[30,50] and this could open new possibilities to fractionate, derivatize
and process lignocellulosic material [48,68]. Imidazolium salt based ILs
have been used by different research groups and these have high po-
tential for lignocellulosic biomass dissolution [85]. This effect was
further enhanced by Mei et al. who used ILs in combination with polar
organic solvents, to significantly enhance the solubility of lig-
nocellulose. When the process was combined with microwave treat-
ment, the resulting cellulose was hydrolyzed at least 22 times faster
than that of untreated rice straw by cellulase from Trichoderma reesei
[86]. In spite of the high potential in enhancing digestibility of cellu-
losic biomass, the application of ILs is still in research stage. For com-
mercial use, the problems associated with the high cost of chemicals
and regeneration of ILs must be resolved.
5.1.1.4. Physio-chemical methods. In physio-chemical pretreatment,
biomass structure is destroyed, and lignin and hemicellulose are
separated by changing the operating conditions – usually pressure
and temperature [71]. The most common method is SE. This is a widely
used thermo-chemo –mechanical process, where the structural
components of biomass are broken down by steam heating (thermo),
shearing due to the sudden decompression and expansion of moisture
(mechano), and hydrolysis of glycosidic bonds (chemical) [30,68]. In
this method, chipped biomass is treated with high-pressure saturated
steam at a desired temperature. The high temperature during
pretreatment initiates autohydrolysis, which causes hemicellulose
hydrolysis, lignin transformation, and decreases cellulose crystallinity
[33,80]. During the decompression, the material is ejected from the
reactor and is recovered in the explosion tank [70]. When the pressure
is suddenly reduced to atmospheric pressure, material undergoes an
explosive breakdown due to moisture evaporation and gas volume
expansion [51].
The SE process can be modified by replacing steam with different
gases like CO2, SO2 and ammonia to enhance the degradation of lignin
and improve the enzymatic hydrolysis [13,75]. The AFEX method is
based on an SE method, where ammonia is added to biomass with
steam. Biomass is heated to moderate temperatures (70–100 °C) and
moderate pressure (150–400 psi) is applied [81]. AFEX results in partial
solubilisation of hemicelluloses and increased accessibility of the cell
wall polysaccharides for enzyme [81]. In addition, rapid expansion of
the ammonia gas causes destruction of lignin-carbohydrate complex
and consequent physical disruption of biomass fibers, leading to in-
creased digestibility of biomass [50]. After pretreatment, ammonia in
the gas phase is recovered and recycled [73,81].
Another modification of SE pretreatment method is CO2 explosion.
In CO2 explosion, supercritical CO2 under pressure and at high tem-
perature is used to enhance the digestibility of lignocellulosic biomass
[30]. CO2 molecules can penetrate into the biomass and when dis-
solved, an acid-catalyzed process is initiated by carbonic acid [50],
which increases dissolution of hemicellulose [30]. CO2 and AFEX pre-
treatment methods require lower temperatures than SE pretreatment
and therefore, they are more cost-effective [71].
In addition to SE, a new approach, NED pretreatment of biomass
was recently proposed [62,87]. The NED pretreatment is based on the
heating of biomass at elevated pressure, which is applied by adding N2
gas. Heating initiates autohydrolysis and at the same time the nitrogen
molecules, which are smaller than the water molecules or gas molecules
used in other pretreatment methods (like CO2 and SO2), can more ef-
fectively penetrate into the cells of biomass. The following explosive
release of pressure causes the dissolved nitrogen gas to expand. This
M. Raud, et al.
opens the cellulosic structure of the biomass and increases the acces-
sible surface area for enzymatic hydrolysis. NED pretreatment method
is economically and environmentally attractive and enables to gain
glucose yields between 278 and 338 g/kg depending on the pressure
[62,87]. However, the method's working principle and the processes
occurring during the pretreatment still need further research.
LHW is a hydrothermal treatment, which does not require rapid
decompression and does not employ any catalyst or chemicals [75]. The
method is similar to SE but uses water in the liquid state at high tem-
peratures. The water and biomass are brought in contact for up to
15 min at temperatures of 170–230 °C. LHW pretreatment results in
hemicellulose hydrolysis and removal of lignin. The cellulose in the
biomass is thereby more accessible while the formation of fermentation
inhibitors at higher temperatures can be avoided [30]. The pretreated
biomass slurry can be divided into solid cellulose-enriched fraction and
a liquid fraction rich in hemicellulose derived sugars [75].
5.1.2. Hydrolysis of cellulose
Processing of pretreated biomass to ethanol consists of three major
units of operations: hydrolysis of cellulose to sugars, fermentation of
sugars to ethanol and product separation/purification. Cellulose hy-
drolysis, also known as saccharification, is the process in which the
cellulose is converted into glucose. This can be done either en-
zymatically by cellulases or chemically by acids [58,69]. In addition to
cellulose, hemicellulose can also be hydrolyzed into its components
(mannan, xylan, glucan, galactan, and arabinan) [57]. The acid hy-
drolysis can be conducted with diluted acids at high temperature
(200–240 °C) or with concentrated acids at lower temperatures. The
latter is considered to be a more effective method as more free sugars
and less inhibitors are produced. However concentrated acid process
requires large amounts of chemicals, which makes it less attractive
[44]. In most cases, the hydrolysis of cellulose and hemicellulose is
carried out using microbial cellulolytic enzymes. Enzymatic hydrolysis
is very mild process, which enables to gain potentially high glucose
yields, and the maintenance costs are low compared to acid or alkaline
hydrolysis [57]. Furthermore, no degradation products of glucose are
formed during the enzymatic hydrolysis [69].
Enzymatic hydrolysis of lignocellulosic biomass is a process, which
includes different biochemical reactions to convert cellulose and
hemicellulose into monosaccharides. Different sets of hydrolytic en-
zymes, such as cellulases, hemicellulases, and accessory enzymes are
required for complete deconstruction of lignocellulosic biomass [88].
For efficient conversion of cellulose into sugars, at least three categories
of enzymes are necessary – endogluconases, exogluconases and β-glu-
cosidases [89]. In the process of hydrolysis, the endoglucanase ran-
domly attacks low crystalline areas in the cellulose fiber to produce
cello-oligosaccharides with free chain ends. At the same time, exoglu-
canases cleave off cellobiose units from the ends of the cello-oligo-
saccharides and generate cellobiose, which is further hydrolyzed by β-
glucosidases to glucose [90,91]. Individual cellulases have very limited
hydrolytic activity. However, as a cellulase mixture, they work sy-
nergistically and thereby, the hydrolytic activity is greater than the sum
of the hydrolytic activities of the individual enzyme [89]. Use of cel-
lulase supplemented with other enzymes can raise the rate of enzymatic
hydrolysis. It is well known that conjugated action of cellulases and
hemicellulases results in a higher ultimate sugar production [52]. Zou
et al. have shown that using specially constructed and statistically op-
timized mixture improved the hydrolytic efficiency and produced twice
higher glucose yield in comparison with crude cellulase preparation
[89].
Cellulolytic enzymes are widespread in nature and are produced by
various plants, insects, and microorganisms [91]. Aerobic fungus Tri-
choderma reesei, which produces an extracellular, stable, and efficient
cellulase enzyme system is the most frequently reported source of cel-
lulases [92] and the most commonly used in lab and pilot-scale bioe-
thanol production process [93,94]. Cellulases are commercially
51
Renewable and Sustainable Energy Reviews 111 (2019) 44–56
available and different studies on cellulolytic enzymes have demon-
strated their biotechnological potential in industrial applications [95].
The commercial enzyme mixtures are mainly supplied bycompanies
Novozyme (Celluclast, Novozyme 188 etc.) and Genencor (Spezyme
and Accelarase 1500 etc.) [96,97].
Factors affecting enzymatic hydrolysis process involve substrate
concentration, cellulase activity and loading, various reaction condi-
tions (temperature, pH and hydrolysis time), and presence of inhibitors
formed during pretreatment [57]. The cellulose properties, which affect
the rate of hydrolysis include: (1) molecular structure of cellulose, (2)
crystallinity of cellulose, (3) surface area of cellulose fiber, (4) degree of
swelling of cellulose fiber, (5) degree of polymerization, and (6) asso-
ciated lignin or other materials [51]. Currently, the most used ways to
enhance the enzyme efficiency and gain higher sugar yields from hy-
drolysis are the optimization of process conditions, application of dif-
ferent enzyme mixtures and additives to improve cellulase performance
[98].
A promising alternative to increase the enzymatic hydrolysis yields
is the addition of different additives, like non-ionic surfactants as these
additives adsorb on the biomass preventing the binding of the hydro-
lytic enzymes and subsequently increasing their effectiveness. Rocha-
Martín et al., 2017 showed that the addition of non-ionic surfactants
and PEG4000 to hydrolysis mixture enabled to increase the release of
glucose and also helped to reduce the liquefaction time [99]. Similar
results were also gained using other surfactants like poly-
vinylpyrrolidone (PVP) and Tween 20 in enzymatic hydrolysis process
[100,101].
5.1.3. Fermentation strategies for bioethanol production
The hydrolyzed biomass containing different sugars can be fer-
mented to ethanol by several microorganisms. However, while the
hexoses (mainly glucose), can be fermented to ethanol by many natu-
rally occurring organisms, the pentoses (xylose and arabinose) can be
fermented to ethanol only by few native strains at relatively low yields
[58]. For commercial bioethanol production the fermenting micro-
organisms should have broad substrates utilization range, high ethanol
yield and productivity, high tolerance to temperature, low pH, in-
hibitors formed in previous process steps and high concentrations of
sugar and ethanol [52,57,94]. The most commonly used industrial
fermentation microorganisms are yeast Saccharomyces cerevisiae and
bacterium Zymomonas mobilis. Unfortunately, these microorganisms
only ferment the hexoses, not the pentoses [69,94,102]. However, sugar
derived from the lignocellulosic biomass, particularly from hardwood
and agricultural waste, contains in addition to glucose 5–20% of the
pentose sugars, like xylose and arabinose [93,102]. The most promising
organisms that have the ability to ferment both pentoses and hexoses
are Pichia stipitis, Candida shehatae, Pachysolan tannophilus and Kluy-
veromyces marixianus. However their efficiencies are lower than that of
S. cerevisiae and Z. mobilis, and they are sensitive to process conditions
(temperature, pH, inhibitors etc.) [57,94].
Production of ethanol from biomass can be done using different
approaches and in Table 3 the comparison of different hydrolysis and
fermentation strategies is outlined. The classic configuration employs
separate enzymatic hydrolysis, which is followed by fermentation in
SHF processes, where the hydrolysis of cellulose and the fermentation
of glucose are carried out in separate units. The major advantage of this
method is that both processes would get optimum conditions - for
cellulose hydrolysis (45–50 °C) and ethanol production (30–37 °C).
However, the process may be affected by the accumulation of sugars
inhibiting the enzyme activity [100,103].
In the SSF, the hydrolysis and fermentation are performed in a
single unit, which enables substantial reductions in equipment, in-
vestment, and operation costs [57,69]. In the SSF process, the sugars
formed from biomass are rapidly fermented into ethanol and thereby,
accumulation of inhibiting sugars can be avoided and thus, higher
ethanol yields with lower amounts of enzyme could be gained [104].
M. Raud, et al. Renewable and Sustainable Energy Reviews 111 (2019) 44–56

The research by Carrillo-Nieves et al. showed that in bioethanol pro-

[57,69,104]
References

[100,103]

[103,106]
duction from mango stem bark residues the SSF strategy was superior to
SHF and achieved higher ethanol yields and concentrations [100]. In

[94]
addition, Dahnum et al. concluded similarly that SSF method was better
than SHF due to faster ethanol production and higher concentrations of

Thermophilic microorganisms have low bioethanol tolerance

produced ethanol. The SHF method produced 4.74% of ethanol in 72 h
Difficulty in optimization of process parameters for both,

however with SSF process, 6.05% of ethanol in was produced in 24 h

Accumulation of sugars inhibiting the enzyme activity

Different assimilation rates of pentoses and hexoses [105]. The main drawback of the SSF process is gaining simultaneously
the optimum temperature conditions for maximum enzyme activity
enzymes and microorganisms at the same time

(50 °C) and yeast viability (28–37 °C). As the enzyme modification
through protein engineering is difficult, the thermotolerant yeast
strains that could efficiently produce ethanol at high temperature are
required [103]. A promising candidate for industrial application is
thermotolerant Kluyveromyces marxianus, which can grow at tempera-
tures as high as 45–52 °C and efficiently produce ethanol at tempera-
tures between 38 °C and 45 °C [106]. Thermophilic cellulolytic anae-
robic bacteria have a distinct advantage over conventional yeasts in
bioethanol production. They are able to use directly variety of in-
Disadvantages

expensive biomass feedstocks and withstand temperature extremes.

However, the low bioethanol tolerance (< 2%, v/v) is a major obstacle
remains

for their industrial exploitation for bioethanol production [94,103].

When SSF includes the co-fermentation of hexose and pentose su-
gars, it is called simultaneous saccharification and co-fermentation
Enzymatic hydrolysis and fermentation function at

Enzymatic and fermentation processes are entirely

Lower equipment requirement than with the SHF

(SSCF). This can be achieved by application of mixed cultures.

Whole substrate is utilized for release of sugars
No accumulation of inhibitory sugar in media

Enhanced efficiency of bioethanol production

However, in case of mixed cultures the microorganisms utilizing

Fewer inhibitory effects during hydrolysis

hexoses grow faster than microorganisms utilizing pentoses and there-

fore, the conversion of hexoses to ethanol is predominant.
Substantially lower production costs
their respective optimum conditions

Alternatively, a single microorganism able to assimilate both hexoses

Less vulnerable to contamination

No cost for enzyme production

and pentoses and enable to gain high conversion and ethanol yield, can
be utilized [94]. In the study by by Chen et al., SSCF bioreactor with
Lower contamination risk

immobilized cultures was used to produce bioethanol from sawdust

Shorter operation time
Lower reactor volume

using direct conversion process. The study showed the synergistic effect
on enzyme activity, when co-cultivation and immobilization of dif-
Easy to operate

ferent cultures of A. niger and T. reesei and Z. mobilis onto carriers and
Advantages

compatible
Low cost

into beads were combined. The immobilization improves the SSCF

process and further lowers the cost in comparison with SHF processes.
However, additional research is required to better understand the
commercialization aspect of this process [107].
Enzymes production, enzymatic hydrolysis and fermentation are carried out in
Saccharification and simultaneous fermentation of released sugars in a single

Saccharification and simultaneous fermentation of released sugars in a single

A step further is CBP, where cellulase production, cellulose hydro-

lysis, and fermentation are carried out in a single step. In the CBP,
bioethanol is produced by a single microbial community and the entire
reactor using yeasts that can assimilate both hexoses and pentoses

process is carried out in one reactor. In addition to single strain pro-

cesses [108], stable mixed cultures that can degrade lignocelluloses
have been used. The thermophilic mixed culture of 5 main strains re-
ported by Lin et al. effectively degraded a diverse range of lig-
Fermentation and hydrolysis are operated separately

nocellulosic materials and produced various biochemical products in-

a single step by a single microbial community

cluding acetic acid, ethanol, and butanol [109]. Svetlitchnyi reported

Comparison of different hydrolysis and fermentation strategies.

the use of extremely thermophilic co-cultures of Caldicellulo siruptor and

Thermo anaerobacter that were capable of converting C6- and C5-sugars
from cellulose and various pretreated lignocellulosic materials into
ethanol, lactate and acetate, while ethanol was the main fermentation
product [110].
The main advantages of this approach are the reduced capital costs,
since only one reactor is required, the costs for enzyme production can
be avoided, whole substrate is utilized for release of sugars and no part
of it goes for cellulase production. Moreover, the enzymatic and fer-
Description

mentation processes are entirely compatible [103,106]. Thereby,

reactor

compared to independent hydrolysis and fermentation steps, sub-

stantially lower production costs and enhanced efficiency of bioethanol
production can be gained [111]. The method has gained attention as a
Fermentation strategy

potential option for low-cost bioethanol production. However, its via-

bility depends on whether a suitable microorganism can be found in
nature or engineered in the laboratory. The microorganisms used in
CBP must be capable of simultaneous cellulose saccharification and
Table 3

SSCF

ethanol production from sugars [112]. Different bacteria and fungi

CBP
SHF

SSF

cultures have been proposed as the primary candidates for CBP research

52
M. Raud, et al.
and some progress has been made in this regard [113,114].
5.2. Liquid fuels from thermochemical pyrolysis process
Pyrolysis processes are distinguished between slow and fast pyr-
olysis. The slow pyrolysis is used mainly for the production of chemicals
and biochar, while fast pyrolysis is more suitable for fuel oil production
[115]. In fast pyrolysis, biomass is heated rapidly in the absence of
oxygen to a temperature around 500 °C [116]. The thermal decom-
position of the biomass produces vapours that are condensed, forming
an organic liquid called FPBO. The production process is close to full
maturity and commercial scale exists in Finland (Fortum) as well as in
the Netherlands (Empyro - BTG-Bioliquids BV) [117]. The FPBO can be
used in heating and power plants and in diesel engine and gas turbine
applications to replace fossil fuels. The usage in district heating plants
to replace the usage of heavy fuel oil has been demonstrated to be a
viable option, both in respect to usability of the fuels as well as the flue
gas emissions [118,119] (Lehto et al., Sippula et al., 2019).
Fast pyrolysis liquid, bio-oil, is formed after cooling and condensing
the vapours from this decomposition process. The liquid has a heating
value about half that of conventional fossil fuel oil [116]. Fast pyrolysis
bio-oil (FPBO) is typically a dark brown, free-flowing liquid composed
of a complex mixture of oxygenated organic compounds [116]. Typical
water contents of FPBO is 20–30% and typical elemental contents is
44–47% of carbon, 6–7% of hydrogen, 46–48% of oxygen and 0-0,2% of
nitrogen [115]. The most prominent organic compounds in the FPBO
include acetic acid, methanol, aldehydes and ketones, cyclopentenones,
furans, alkyl-phenols, alkylmethoxyphenols, anhydrosugars, oligomeric
sugars and water-insoluble lignin-derived compounds [117]. FPBO is
miscible with polar solvents such as methanol, acetone etc. because of
its high oxygen [116]. The high oxygen content also leads to high
viscosities and boiling points, and relatively poor chemical stability,
which is one of the major challenges in storing of the substance [115].
Characteristics of the FPBO can vary considerably according to the
feedstock material, the process parameters and the conditions at the
liquid collection (especially temperature of liquid collection system and
the method of collection). The oil pH is typically around 2.5, ash con-
tent is around 0.1% and pour point typically below −20 °C [117].
Higher heating value (moisture free) is typically around 22.5 MJ/kg
and the heating value as produced is around 17 MJ/kg, depending on
oil moisture content [115]. Some of these characteristics have strong
effects on the usability of the oils. Although FPBO has been successfully
demonstrated in commercial utilization in boiler applications, there are
still challenges and development needs to improve its utilization in
energy productions, including:
- Suspended char in the oil can cause e.g. erosion and burner equip-
ment blockage, boiler deposits and high emissions [115]. e.g. oil
filtration can be used to solve these problems.
- Alkali metals present in the fuel can form deposits in the boiler and
e.g. oil filtration, oil upgrading, as well as feedstock material control
can be used as a solutions.
- Low oil pH can cause corrosion in the fuel delivery system, which
can be prevented by careful material selections [118].
- Polymers can have an effect on sealing rings and gaskets, which also
have to be taken into account when choosing the materials.
- High temperature sensitivity can cause liquid decomposition on hot
surfaces and can be solved using appropriate cooling facilities. High
viscosity can cause high pressure drops in pipelines and an addition
of water/ethanol/methanol or other solvent can be a solution for
that problem.
- The oil has typically a high fuel water content which affects visc-
osity, heating value, density, stability, pH and homogeneity [115].
Too high oil water content may cause difficulties in ignition, flame
stability and controllability of the combustion process during op-
eration. Thus, combustion of fast pyrolysis bio-oil requires burner
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Renewable and Sustainable Energy Reviews 111 (2019) 44–56
equipments specifically designed for the fuel [118].
- Inhomogeneous fuel can cause problems in the storing, transporting
and combustion of the fuel. Pyrolysis bio-oils tend to phase-separate
and form layers during storing. The oil tanks, transport lines and
burners need to be specifically designed for pyrolysis bio-oil to
comply with the properties and inhomogeneity of the fuel.
The physicochemical properties of bio-oil could be improved by
physical upgrading processes or chemical and thermocatalytic opera-
tions. Mostafazadeh et al. (2018) have reviewed the alternatives in
upgrading of different bio-oils, including both chemical and physical
upgrading methods [120]. According to Bridgwater (2012), the most
relevant physcial upgrading methods for FPBO are filtration, solvent
addition and emulsification [121]. Potential catalytic upgrading
methods for FPBO are e.g. hydrotreatment, catalytic cracking and es-
terification. However, FPBO upgrading methods are not yet in com-
mercial use. So far bubbling fluidized bed and circulating fluidized bed
technologies have been used for commercial production of FPBO [122].
Although the production of FPBO has numerous challenges to meet,
biomass can still be considered as a potential resource for producing
qualified FPBO using fast pyrolysis followed by proper upgrading
methods [120]. It can be concluded that the highest potential in FPBO
utilization is currently in replacing fossil fuel oil in industrial scale
heating and power boilers. In addition, FPBO has potential to be uti-
lized in stationary heat and power producing large scale diesel engines
and gas turbines [117]. Longer term goal is to use FPBO as co-feedstock
in oil refineries for the production of transportation fuels and/or dif-
ferent petrochemical feedstocks. (e.g. Solantausta et al., 2012). Re-
cently commercialized facilities for FPBO have been opened in Joensuu
(Finland), where Fortum Ltd opened FPBO production plant [123] and
also Empyro has opened an FPBO production plant in Netherlands
[124].
6. Improving the process efficiency of bioethanol production:
potentials and challenges
Tremendous advances have already occurred in the lignocellulose
industry for the production of biofuels from lignocellulosic biomass.
According to the online database of facilities for the production of ad-
vanced liquid and gaseous biofuels for transport, a number of projects
with advanced biofuel production facilities at different levels and scales
are under development in several continents worldwide. Lignocellulosic
biofuel demonstration and full scale commercial plants are under con-
struction. Many of them are already operational in Europe, North, - and
South America and Asia [125]. In spite of the apparent success, several
big bioethanol plants have been closed and projects have been put on
hold [126,127].
Although the first bioethanol production facilities are operational,
the processing technologies are in developmental stage and therefore,
progress needs to be made to improve the overall process efficiency
[14]. One of the main problems is the relatively high capital and pro-
cessing costs [128] since the advanced technologies and facilities are
required to aid the conversion process [31]. The price will come down
when production is moved to larger scale production facilities and more
experience is gained. Currently, the research and development in this
field has been funded from a number of sources, both public and pri-
vate, which include venture capital funds and internal funds from large
corporations [129]. This support has helped to move it further however,
more support is needed to bring this technology into maturity.
Even though the production of liquid biofuels from lignocellulosic
biomass show promise with good production yields in laboratory con-
ditions, there are still several technical barriers hindering the large-
scale and cost-effective production of cellulosic biofuels [14,31,128].
Due to the large number of individual processes in the overall
conversion of lignocellulosic biomass, there is a considerable potential
for process integration [14]. Possibilities for the cost reduction include
M. Raud, et al.
more effective pretreatment [11,14,82], reducing the cost of cellulase
by improving enzyme efficiency [11,14,68,82], hydrolysis of both
hemicelluloses and cellulose to sugars [11], use of both six-carbon and
five-carbon sugars in fermentation [11,14], and process integration for
reducing capital and energy costs [11]. The overall revenues from the
process could be increased with production of co-products during the
bioethanol production [14,58] and utilization of the residual material -
residual lignin, unreacted cellulose and hemicellulose, enzymes, yeast,
and other components, for production of biogas in anaerobic digestion
process [130]. Such a combined production aims at gaining higher total
energy yield than with just ethanol production from hexoses and pen-
toses [130].
It is estimated that the pretreatment step constitutes 40% of pro-
cessing costs in bioethanol production process. Therefore, it is vital to
select the optimal pretreatment technology, to gain the cost-effective
bioethanol production process from lignocellulosic biomass [73]. In
case of various chemical pretreatment methods, the key issue is the
chemical recovery [68] and economical usage of chemicals due to high
cost of chemicals. Chemical free processes, like SE pretreatment, have
more potential due to reduction of cost by elimination of chemicals.
Currently, there is no ideal pretreatment method that can be adopted
for all biomass types. However, combination of different methods must
be applied to find the suitable process for different types of biomass
[52].
Enzymatic hydrolysis is one of the most challenging steps for the
second generation biofuel commercialization and accounts for 25–30%
of the operational costs [99]. Therefore, the research and development
on cost-effective enzyme cocktails are also one of the major challenges
in biomass hydrolysis. In order to improve the economic efficiency of
lignocellulosic biofuel production, all biomass components and also
residual biomass should be utilized. Therefore, highly effective bioe-
thanol process is required for the complete hydrolysis of both cellulose
and hemicellulose to sugars with minimum sugar losses and efficient
fermentation of all sugars to ethanol [102]. The efficient degradation of
hemicellulose to pentoses and hexoses requires the synergistic action of
many enzymes including hemicellulases [93]. In addition, to fully uti-
lize biomass, the hemicellulose hydrolysates must also be fermented
and utilized [111]. However, when using Saccharomyces cerevisiae for
ethanol fermentation, only hexoses will be fermented to ethanol, and
the pentoses together with proteins and other carbon sources will re-
main unused in residual biomass [102]. Various xylose fermenting
fungal and recombinant bacteria have been reported. Most of them
produce only low ethanol yields and not all are capable of adapting to
fermentation process conditions. Therefore, it is vital to research and
develop genetically modified fermentative and cellulolytic micro-
organisms and co-culture systems to increase ethanol yield and pro-
ductivity [47].
After bioethanol production, a vast amount of biomass, mainly re-
sidual lignin, unreacted cellulose and hemicellulose, enzymes, yeast,
and other components, is left as process waste. To fully utilize the
biomass, the residual material should also be used in a separate process,
like anaerobic digestion (AD) [130] or for heat- and energy production.
One of the benefits of co-production is that enzymes and yeast added
during ethanol production can be converted to methane [130].
A more advanced approach is through integrated biorefinery. This
approach includes integration of biofuel production with other pro-
duction facilities, which use biomass or process residues to make dif-
ferent co-products like biofuel, bio-chemicals, fertilizer, heat and en-
ergy etc. [131]. The economic benefit of this approach is gained when,
in addition to biofuels, high-value but low-volume co-products are also
produced. This way a broader variety of biomass feedstocks could be
used in a more efficient way than current biofuel production units, and
additionally, competition among different uses of biomass reduces
[132]. Initially, the most likely co-products will be fuel, heat and
electricity; however, in future biorefineries, biofuel technology will
form the basis for the sustainable production of different commodity
54
Renewable and Sustainable Energy Reviews 111 (2019) 44–56
chemicals and materials [102].
Besides having several benefits, the production and utilization of
biofuels has several challenges. An improved biomass waste collection
network and their storage is the main challenge for establishment of
commercial biofuel plant. A strong policy is needed for organic waste
collection and blending of biofuels at higher rate. The technological
improvement could help improve the system efficiency and provide
value-added co-products, which will reduce the overall production cost
[8].
7. Conclusions
In recent decades, more attention is being devoted to climate issues
and sustainability in energy sector. In order to decrease the dependence
on fossil fuels by increasing the use of biofuels many different legisla-
tive actions and measures have been adopted at both local and inter-
national levels. In this context, applications employing lignocellulosic
biomass as a cheap and widely available substrate for biofuel produc-
tion have received wide attention. Many modern technologies are
available that enable to produce liquid biofuels from lignocellulosic
biomass. Bioethanol production using microbial fermentation, and bio-
oil (FPBO) production using a pyrolysis process of biomass are among
the technologies that are widely practised.
Even though the liquid biofuel production from lignocellulosic
biomass shows high production yields, there are several challenges that
exist before its commercialization. The production technologies are
close to full maturity for the thermochemical biofuels and in early
commercial prototype phase for bioethanol production. Nevertheless,
further research and development is needed for the thermochemical
biofuels on the material selection for different parts in production and
instrumentation to make them more resistant to harsh process condi-
tions. Also, extensive research has been carried out to make different
bioethanol process steps more efficient – development of various pre-
treatment methods, enzyme development for enhanced hydrolysis, re-
search for more tolerant yeasts capable of fermenting different sugars.
Value addition and perspective, cost saving measures can be achieved
with the application of different combined processing technologies such
as SSF, SSCF and CBP and by using residues from bioethanol process to
produce additional products like bio-chemicals, fertilizer, heat and
energy by applying integrated biorefinery approach.
Acknowledgements
We gratefully acknowledge the financial support of the European
Regional Development Fund via the Mobilitas Pluss (project MOBERA2)
of the Estonian Research Council and the Academy of Finland through
the INDO–NORDEN project (Holistic processes and practices for clean
energy in strengthening bioeconomic strategies, 311970).
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