fermentation

Review
Recent Developments and Current Status of Commercial
Production of Fuel Ethanol
Tuan-Dung Hoang 1, * and Nhuan Nghiem 2

1 Department of Environmental Security and Technology, Hanoi School of Business and Management, Vietnam
National University, 144-Xuan Thuy Street, Hanoi 10000, Vietnam
2 Biosystems Engineering Program, Department of Environmental Engineering and Earth Sciences, Clemson
University, Clemson, SC 29631, USA; [email protected]
* Correspondence: [email protected] or [email protected]; Tel.: +84-936414637

Abstract: Ethanol produced from various biobased sources (bioethanol) has been gaining high
attention lately due to its potential to cut down net emissions of carbon dioxide while reducing
burgeoning world dependence on fossil fuels. Global ethanol production has increased more than six-
fold from 18 billion liters at the turn of the century to 110 billion liters in 2019, only to fall to 98.6 billion
liters in 2020 due to the pandemic. Sugar cane and corn have been used as the major feedstocks
for ethanol production. Lignocellulosic biomass has recently been considered as another potential
feedstock due to its non-food competing status and its availability in very large quantities. This
paper reviews recent developments and current status of commercial production of ethanol across
the world with a focus on the technological aspects. The review includes the ethanol production
processes used for each type of feedstock, both currently practiced at commercial scale and still under
developments, and current production trends in various regions and countries in the world.





 Keywords: fuel ethanol; renewable energy; biobased feedstocks; lignocellulosic biomass;
Citation: Hoang, T.-D.; Nghiem, N. fermentation process; processing options; commercialization; production status; climate change;
Recent Developments and Current environmental security
Status of Commercial Production of
Fuel Ethanol. Fermentation 2021, 7,
314. https://doi.org/10.3390/
fermentation7040314 1. Introduction
Ethanol (C2 H6 O) is a simple liquid alcohol that is formed from the fermentation of sug-
Academic Editor: Diomi Mamma
ars in their natural occurrences or being derived from starch-rich grains or lignocellulosic
feedstocks. Ethanol is also called ethyl alcohol, grain alcohol, or simply alcohol, and is used
Received: 8 November 2021
as a disinfectant, an organic solvent, a chemical feedstock, and a transportation liquid fuel.
Accepted: 10 December 2021
Published: 15 December 2021
Ethanol is produced in various countries in the world and its global quantity has changed
from 110 billion liters in 2019 to 98.6 billion liters in 2020 due to the pandemic [1,2]. When
ethanol is blended with 95% gasoline it can reduce about 90% CO2 and 60–80% SO2 [3].
Publisher’s Note: MDPI stays neutral
with regard to jurisdictional claims in
This helps the world to solve some of the problems of air pollution, reduces the levels of
published maps and institutional affil-
greenhouse gases that are causing climate change and maintains environmental security.
iations. Currently, ethanol is produced commercially from a variety of feedstocks via fermentation
where the yeast Saccharomyces cerevisiae is utilized to ferment the sugars derived from the
starch in corn and other grains or those that already are present in sugar cane and sugar
beets [4].
Crude oil and natural gas are traditionally the main raw materials for producing
Copyright: © 2021 by the authors.
fuels and industrial chemicals. However, human dependency on fossil fuels has become
Licensee MDPI, Basel, Switzerland.
This article is an open access article
a critical issue worldwide and it is essential to explore new sustainable and alternative
distributed under the terms and
sources to solve the world’s environmental problems [5]. Ethanol is an important source of
conditions of the Creative Commons biofuels and the ethanol development and implementation is within the scope of circular
Attribution (CC BY) license (https:// bioeconomy since biofuels can substitute for fossil carbon by bio-based carbon in biomass
creativecommons.org/licenses/by/ from agriculture, forestry and municipal wastes. In terms of sustainability of bioethanol
4.0/). production compared to fossil-based fuels, ethanol development is a complementary

Fermentation 2021, 7, 314. https://doi.org/10.3390/fermentation7040314 https://www.mdpi.com/journal/fermentation

Fermentation 2021, 7, 314 2 of 18

approach and based on improved resources with higher eco-efficiency and reduced GHGs
footprint [6]. Furthermore, the increase of liquid fuels supplying the energy demand with
regards to economic and environmental concerns make ethanol a good energy alternative
for many countries.
There are commonly three groups of materials that can be used for manufacturing
ethanol, which are: (a) feedstocks which contain substantial amounts of readily fermentable
sugars, (b) starches and fructans, and (c) cellulosic materials. However, in-depth discus-
sions on the different feedstocks vis-à-vis production technologies and current development
of ethanol worldwide are still lacking. The main objective of this review is to investigate
the recent development of ethanol production in countries with significant levels of ethanol
outputs. The review also examines the current technologies for ethanol production and
those being developed toward commercial implementation.

2. Production Technologies
Three types of feedstock can be used for ethanol production. These include sugar-
based feedstocks (sugar cane, sugar beet), starched-based feedstocks (corn, barley, wheat,
other grains), and lignocellulosic feedstocks (agricultural residues, forest residues, ded-
icated energy crops, municipal solid wastes). The ethanol produced from sugar-based
and starch-based feedstocks is referred to as first-generation ethanol whereas the ethanol
produced from biomass feedstocks is referred to as second-generation ethanol.

2.1. Sugar-Based Feedstocks

The juice extracted from sugar cane has been used for ethanol production. Part of
the juice is used for sugar manufacture in an adjacent sugar manufacturing plant and the
remaining is used for ethanol production such as the case of Brazil, which is the second
largest ethanol producer in the world. Juice extraction can be practiced through either
crushing (roll mills) or diffusion (diffuser). The decision to produce sugar and/or ethanol
is made by individual plants from harvest to harvest. This is a very important issue since
once the producing units adjust their plants to produce a set ratio of sugar/ethanol in a
given year, it is very difficult to change during the harvest season [7]. Bagasse, which is the
solid residue obtained after juice extraction, has been combusted to generate energy for
internal uses, but also has the potential as a lignocellulosic feedstock for additional ethanol
production. Molasses, which is a by-product of sugar cane processing, can also be used for
ethanol production.
Typically, sugarcane contains 12–17% total sugars on a wet weight basis with 68–72%
moisture. About 90% of the sugars are sucrose with glucose and fructose making up the
balance. All these three sugars are readily fermentable by the yeast S. cerevisiae to produce
ethanol. During the fermentation process, the yeast produces the enzyme invertase and
uses it to convert sucrose to glucose and fructose. Ethanol production by S. cerevisiae is
carried out via the glycolytic pathway (also known as the Embden-Meyerhof-Parnas or
EMP pathway). In the simplest form, production of ethanol from glucose can be expressed
by the following equation:

C6 H12 O6 + 2 Pi + 2 ADP → 2 C2 H5 OH + 2 CO2 + 2 ATP + 2 H2 O

glucose → 2 ethanol + 2 carbon dioxide + energy

According to the above equation, the maximum theoretical yield of ethanol produced
from 1 g of glucose is 0.511 g ethanol. In practice, it is observed that the actual yield is
always lower since not all of the glucose consumed is converted to ethanol [8].
The fermentation process can be either batch/fedbatch or continuous. The Melle-
Boinot batch process, which was developed in the 19300 s, is most commonly used. Its
main characteristic is total yeast recycle, normally by centrifugation. The very high yeast
densities (10–14% w/v), which are obtained by yeast recycle, allow reduced growth, high
ethanol yield and very short fermentation time (6–10 h). The continuous process with cell
Fermentation 2021, 7, 314 3 of 18

recycle was developed in the 19800 s to replace the batch process [9]. The majority of the
commercial plants in Brazil, however, still employ the batch/fed-batch process. The ethanol
produced is recovered by distillation followed by molecular sieve or other dehydration
technology to produce anhydrous ethanol (>99.5%). The stream coming out at the bottom
of the distillation columns is called vinasse. Vinasse normally is spread on the sugar cane
fields for use as irrigation water and fertilizer. However, this practice may cause extensive
pollution of the environment due to the high organic contents, dark color, dissolved solids
and other compounds that are toxic under certain conditions. Methods for treatment of
vinasse prior to their disposal have been developed [10].
Sugar beet is grown in regions with temperate climates and is one of the main feed-
stocks used for ethanol production in Europe. Sugar beet has a conical, white and fleshy
root, which serves as a reservoir for sugar. The root contains about 75% water and be-
tween 15% and 21% total sugars (wet weight basis). Due to the similarities in chemical
compositions of sugar solutions extracted from sugar cane and sugar beet, those obtained
from sugar beet also can be fermented to ethanol by industrial yeast at high efficiencies.
The fermentation processes are very similar in both cases. Similar to sugar cane, ethanol
production from sugar beet can be integrated with sugar production.

2.2. Starch-Based Feedstocks

Corn is used almost exclusively for ethanol production in the United States where
ethanol is produced from corn by either the wet-milling or dry-grind process. The United
States is the largest producer of ethanol in the world where the majority of the commercial
ethanol plants use the dry-grind process. Ethanol can also be produced from other grains
by the dry-grind process.
In the dry-grind process, after grinding, water and a thermostable α-amylase are
added to the ground corn. In the next step, which is called pre-liquefaction, the slurry then
is brought up to 60–70 ◦ C (warm cook) or 80–90 ◦ C (hot cook). The slurry, which is referred
to as the corn mash, is held at these temperatures for about 30–45 min. The swelling and
hydration of the starch granules cause dramatic increase of the slurry viscosity and loss
of crystallinity of the granule structures. In the next step, which is called liquefaction,
the mash is maintained at 85–95 ◦ C for a period of time or forced through a continuous
jet cooker at 140–150 ◦ C. At the end of the liquefaction, starch is hydrolyzed to short-
chain dextrins (two to four glucose units). The temperature of the mash is lowered to
32 ◦ C and the pH adjusted to approximately 4.5. The mash then is placed in a fermentor.
Glucoamylase and the yeast culture from the yeast propagation tank are also added. Urea
may be added as a nitrogen source. The process combining enzymatic hydrolysis and
fermentation is called simultaneous saccharification and fermentation (SSF). The SSF is a
batch process which typically is run for about 50–60 h. The final ethanol concentration is
about 15 % (v/v). Commonly used commercial glucoamylase formulations also contain
proteases. These enzymes break down organic nitrogen sources in the corn slurry and
release additional nutrients, which help to improve fermentation efficiency and ethanol
yield. The ethanol produced is recovered by distillation followed by molecular sieve as
described previously. The bottom stream from the distillation columns is called stillage.
This stream is dried to a moisture content of about 10% to produce distillers dried grains
with solubles (DDGS), which are sold or used as animal feed. DDGS is the most important
co-product in a dry-grind ethanol plant. The other co-products are the carbon dioxide
produced in the fermentation process and corn oil. The dry-grind process is shown in
Figure 1.
In the wet-milling process, corn grains are soaked in water containing 0.1 to 0.2% SO2
at 52 ◦ C (125 ◦ F) for 24 to 40 h. After this so-called steeping step, the softened grains are
then ground gently to break up the kernels. The less dense germs, which contain about
40 to 50% oil, are recovered in a hydrocyclone system and processed into value-added
co-products. After the germs are removed, the slurry was subjected to further grinding to
loosen the starch and gluten from the remaining fiber. The slurry is screened to remove
Fermentation 2021, 7, 314 4 of 18

the fiber, which then is washed and pressed to about 60% moisture. The water from the
steeping step, which is called light steep water (LSW), is concentrated in evaporators to
produce heavy steep water (HSW). The HSW is dried together with the recovered fiber and
the resultant product and is sold to the livestock industry as corn gluten feed, which typically
contains about 21% proteins. After fiber removal, the gluten, which is lighter, is separated
from the starch in a centrifuge. The recovered gluten is concentrated from, cooled to 35 ◦ C
and filtered on a rotary vacuum filter to produce a gluten cake of 60% moisture. The cake
is dried to 10% moisture to produce corn gluten meal. This feed product contains at 4least
Fermentation 2021, 7, x FOR PEER REVIEW of 19
60% protein and 1% fat, and up to 3% fiber. It is used in poultry feed because of its high
protein and xanthophyll and low fiber contents.

Glucoamylase
Corn α-amylase Yeast
CO2

Simultaneous
Grinding Liquefaction Saccharification &
Fermentation (SSF)

Distillation

Anhydrous
Dehydration
ethanol

Corn
Drying
distillers oil

DDGS + Other
distillers grains
products

Figure1.1.The
Figure Thedry-grind
dry-grindprocess
processfor
forethanol
ethanolproduction
productionfrom
fromcorn.
corn.

The starch is washed and processed through a series of up to 14 hydrocyclones to

In the wet-milling process, corn grains are soaked in water containing 0.1 to 0.2% SO 2

remove impurities. The final product, which is 99.5% pure starch, then is used for ethanol
at 52 °C (125 °F) for 24 to 40 h. After this so-called steeping step, the softened grains are
fermentation in dedicated plants or processed further to produce modified starch, corn
then ground gently to break up the kernels. The less dense germs, which contain about 40
syrup (CS) and high fructose corn syrup (HFCS) in integrated plants. The wet-milling
to 50% oil, are recovered in a hydrocyclone system and processed into value-added co-
process is shown in Figure 2.
products. After the germs are removed, the slurry was subjected to further grinding to
The hydrolysis of starch to produce glucose can be expressed by the following equa-
loosen the starch and gluten from the remaining fiber. The slurry is screened to remove
tion:
the fiber, which then is washed and pressed to about 60% moisture. The water from the
(C6 H10 O5 )n + n H2 O → n C6 H12 O6
steeping step, which is called light steep water (LSW), is concentrated in evaporators to
where
producen is heavy
the number
steep of glucose
water (HSW).residues in theisstarch
The HSW driedmolecule.
together with the recovered fiber
andInthea starch molecule,
resultant productnandis a is
very
soldlarge number.
to the The
livestock theoretical
industry yield
as corn or conversion
gluten feed, which
factor
typically contains about 21% proteins. After fiber removal, the gluten, which isethanol
for glucose is 1.11 g glucose/g starch. In addition, the theoretical yield for lighter,isis
0.57 g ethanol/g starch.
separated from the starch in a centrifuge. The recovered gluten is concentrated from,
cooled to 35 °C and filtered on a rotary vacuum filter to produce a gluten cake of 60%
moisture. The cake is dried to 10% moisture to produce corn gluten meal. This feed product
contains at least 60% protein and 1% fat, and up to 3% fiber. It is used in poultry feed
because of its high protein and xanthophyll and low fiber contents.
The starch is washed and processed through a series of up to 14 hydrocyclones to
remove impurities. The final product, which is 99.5% pure starch, then is used for ethanol
fermentation in dedicated plants or processed further to produce modified starch, corn
Fermentation 7, 3147, x FOR PEER REVIEW
2021, 2021,
Fermentation 5 of 518of 19

Corn

α-amylase
Germ/Fiber Gluten Glucoamylase
Steeping
Removal Separation

Corn syrup
Liquefaction and
Corn oil + Corn Corn gluten High fructose
Saccharification
germ meal meal corn syrup
Yeast
CO2
Drying
Fermentation

Corn gluten
feed
Distillation

Anhydrous
Dehydration
ethanol

Figure 2. The wet-milling process for ethanol production from corn.

Figure 2. The wet-milling process for ethanol production from corn.

The
In the hydrolysis
United States,ofit starch to produce
is commonly glucose
accepted thatcan
onebe expressed
bushel of corn by(56thelbs
following
or 25.4 kg)equa-
tion:
of about 15% moisture processed by the dry-grind process will produce 2.9 gallons (11.0 L)
denatured ethanol, 15.2 lbs (6.9 kg) (Cdistillers grains animal feed (10% moisture), 0.8 lbs
6H10O5)n + n H2O→n C6H12O6
(0.36 kg) distillers corn oil, and 1.1 lbs (0.5 kg) captured carbon dioxide [11].
where
Manynefforts
is the have
number beenof made
glucose to residues
improvein the starch
ethanol molecule.
production with the focus on the
dry-grindInprocess.
a starchThese
molecule, n is a very
improvements arelarge number.
discussed The below.
briefly theoretical
Syngentayielddeveloped
or conversion
the Enogen ® corn
factor for glucose is 1.11
hybrids g glucose/g
containing starch. Inwhich
α-amylase, addition,
helpstheto theoretical yield for of
reduce the viscosity ethanol
the is
corn0.57
mashg ethanol/g starch.
and reduce/eliminate the need of externally added α-amylase [12].
• In the United
Lallemand, States, it iswith
in collaboration commonly accepted that
other companies, one bushel
developed of corn (56
the TransFerm ® lbs or 25.4
Yield+
kg) of about 15% moisture processed by the dry-grind process
S. cerevisiae, which produces the glucoamylase needed for starch hydrolysis and will produce 2.9 gallons
(11.0 L) denatured
improves ethanol ethanol,
yield up 15.2to 4% lbsas(6.9 kg) distillers
a result grains animal
of 30% reduction feed (10%
in glycerol moisture),
synthesis.
0.8 lbs (0.36 kg) distillers corn oil, and 1.1 lbs (0.5 kg) captured
The reduced glycerol levels are not expected to have an adverse effect on the yeast carbon dioxide [11].
Manyofefforts
tolerance osmotic have been
stress madeby
caused to ethanol
improve[13].ethanol production
The newer strain,with the focus
TransFerm on the
CV5,
dry-grind process. These improvements are discussed briefly
is a genetically modified yeast strain that produces high levels of glucoamylase andbelow. Syngenta developed
the Enogen
trehalase ® corn hybrids containing α-amylase, which helps to reduce the viscosity of
expression, which can meet between 80% and 100% of the enzymes required
the corn mash and
in fermentation reduce/eliminate the need of externally added α-amylase [12].
[14].
• •Novozymes
Lallemand, developed the glucoamylase-producing
in collaboration with other companies, Innova
developed S. cerevisiae,
Drive the TransFerm which
® Yield+

could tolerate 98 ◦ F (36.7 ◦ C), 37% solid loading, and 6 g/L organic acids [15]. Other
S. cerevisiae, which produces the glucoamylase needed for starch hydrolysis and im-
strains of theethanol
proves Innovayield
productup toseries
4% asalso have been
a result developed
of 30% reduction within improved tolerance The
glycerol synthesis.
of heat
reduced 40 ◦ C) and
(up toglycerol resistance
levels are not to high levels
expected of solids
to have and high
an adverse concentrations
effect on the yeastof toler-
glucose,
ance ethanol
of osmoticandstress
organic acidsby
caused [16].
ethanol [13]. The newer strain, TransFerm CV5, is a
genetically
Several companies,modifiedwhich yeast
includestrainPoet
that(formerly
produces Broin),
high levels
Cerealof glucoamylase
Process Technolo- and tre-
halase
gies (CPT), expression,
Buhler, Renessen, which
and canFWS meet between 80%
Technologies, haveand 100% of the
developed dryenzymes required
fractionation
processes.inAll
fermentation [14].
dry corn fractionation processes are fundamentally similar. Prior to the con-
•
ventional Novozymes developed
dry-grind process, corn isthe glucoamylase-producing
ground and subsequently fractionatedInnova into Drive S. cerevisiae,
fermentable
and non-fermentable fractions.
which could tolerate 98 Only
°F (36.7 the°C),
fermentable
37% solid fraction
loading, is used
and 6 g/Lin organic
the dry-grind
acids [15].
process for ethanol production. The non-fermentable fractions are
Other strains of the Innova product series also have been developed with improved separated by various
methodstolerance
and recovered
of heatas(up brantoand
40 °C) germandco-products,
resistance towhichhigh are sold
levels of for use and
solids in animal
high con-
feeds. The DDGS obtained
centrations of glucose, afterethanol
ethanol andrecovery
organichas higher
acids [16]. protein, but lower oil and
Fermentation 2021, 7, 314 6 of 18

fiber contents compared to the product obtained in a traditional dry-grind process without
corn fractionation [8].
• D3MAX developed a process for conversion of corn fiber and the unconverted starch
in the stillage to additional ethanol. The D3MAX process is a “bolt-on” process, which
means it can be added to an existing dry-grind ethanol plant. The key step of this
process is the pretreatment of the wet cake obtained by centrifugation of the stillage
with dilute acid. The pretreated wet cake then is subjected to enzymatic hydrolysis
and fermentation, which uses a genetically modified yeast capable of utilizing both
glucose and xylose for ethanol production [17].
• Quad County Corn processors (QCCP) developed the Cellerate process, which is
also a bolt-on process. This process is very similar to the D3MAX process, except
that the whole stillage rather than the wet cake is pretreated with dilute acid prior to
enzymatic hydrolysis and fermentation. It was reported that when Enogen® corn is
used (in collaboration with Syngenta), compared to the traditional dry-grind process
using regular corn, the Cellerate process resulted in 6% increase in ethanol yield, 15%
increase in throughput, and 20% reduction in energy consumption, and produced
1.6 lbs (0.73 kg) corn oil per bushel plus a DDGS with higher protein and lower
fiber [18].
• Edeniq developed the Intellulose® process using their proprietary enzyme mixtures
to produce ethanol from the previously unconverted starch and fibers. It was reported
that the technology resulted in a 2–4.5% increase in ethanol production. Edeniq also
developed an analytical technique to directly measure ethanol production from the
lignocellulosic fractions in the corn kernel [19,20].

2.3. Lignocellulosic Feedstocks

Lignocellulosic feedstocks are also referred to as lignocellulosic biomass or simply
biomass. In this review, the term biomass is used. Biomass consists of three main compo-
nents, which are cellulose, hemicellulose, and lignin. The compositions of representative
biomass feedstocks and the theoretical ethanol yield by the biochemical conversion are
summarized in Table 1.

Table 1. Compositions (dry basis) of representative biomass feedstocks and theoretical ethanol yield by the biochemical conversion [8].

Lignocellulosic
Cellulose (%) Hemicellulose (%) Lignin (%) Theoretical Ethanol Yield (L/MT)
Feedstocks
Forest products
Hardwood 46.2 29.2 22.0 546
Softwood 41.2 26.8 29.8 493
Woody energy crops
Willow 42.5 22.0 26.0 467
Eucalyptus 54.1 18.4 21.5 524
Poplar 52.1 27.5 15.9 576
Pine 46.0 25.5 20.0 518
Agricultural residues
Corn stover 35.2 25.1 23.7 437
Rice straw 43.4 27.9 17.2 517
Barley straw 41.0 26.6 21.3 490
Wheat straw 37.0 26.5 14.0 460
Sugarcane bagasse 41.6 25.1 20.3 483

There are two options for the conversion of biomass to ethanol. In the first option,
cellulose and hemicellulose are hydrolyzed to fermentable sugars, which are subsequently
fermented to produce ethanol. In the second option, the biomass is taken through a process
called gasification. In this process, the biomass is heated with no oxygen or with oxygen
significantly below the normally required for complete combustion. The product is a gas
Fermentation 2021, 7, 314 7 of 18

which
Fermentation 2021, 7, x FOR PEER REVIEW is mostly CO and H2 . The gaseous product is called synthesis gas or syngas, which 7 of 19
can be converted to ethanol via either fermentation or chemical catalysis processes. The
first option normally is referred to as the sugar platform and the second as the syngas
platform.
Table 1. Compositions (dry basis) of representative biomass feedstocks and theoretical ethanol yield
In the sugar platform, only the carbohydrate components of biomass are used for
by the biochemical conversion [8].
ethanol production, whereas lignin is considered as a waste and normally is burned
to generate energy. In the syngas
Lignocellulosic Celluloseplatform, all three Lignin
Hemicellulose components are used for
Theoretical ethanol
Ethanol
production. The key steps of the
Feedstocks (%)sugar platform
(%) and the syngas platform
(%) are shown in
Yield (L/MT)
Figures 3 andproducts
Forest 4, respectively.
Hydrolysis
Hardwood of cellulose and46.2hemicellulose to generate
29.2 fermentable sugars
22.0 546 for ethanol
fermentation can
Softwood be performed with
41.2 either chemicals
26.8 or enzymes.
29.8 The chemicals
493 used for
the hydrolysis of cellulose and hemicellulose include concentrated (>70%) H2 SO4 , which
Woody energy crops
is used in the Arkenol process [21] and the Biosulfurol process [22], supercritical fluid,
Willow 42.5 22.0 26.0 467
which is used in the Renmatix process [23], and γ-valerolactone (GVL), which is used in
Eucalyptus
the GlucanBio process [24]. 54.1 18.4 21.5 524
When concentrated H2 SO4 was used for hydrolysis, the sugar product576
Poplar 52.1 27.5 15.9 was a mixture
of glucose andPinexylose whereas in 46.0
the other two25.5
cases, two20.0
separate streams518
that contained
Agricultural
mostly glucose andresidues
xylose, respectively, were formed. The fractionation of the two car-
bohydrate Corn stover would allow
fractions 35.2the use of 25.1
the C6 sugar stream for ethanol
23.7 437production
by the native S.
Rice strawceresiae strains and
43.4 the C5 sugar
27.9 stream for
17.2 production of higher-value
517
co-products
Barleysuch
strawas xylitol and 41.0
astaxanthin [25].
26.6 21.3 490
Due to the
Wheat strawrigid structure of
37.0biomass, which
26.5 impedes
14.0enzymatic attacks,
460 a process
called pretreatment
Sugarcane bagasse is required before
41.6 the cellulose
25.1 and hemicellulose
20.3 components
483 of
biomass can be effectively hydrolyzed by enzymes. In the pretreatment process the rigid
structure of biomass is opened up and amorphous regions are created. The results of the
In the sugar platform, only the carbohydrate components of biomass are used for
pretreatment process are improvements of the rates of the subsequent enzymatic hydrolysis
ethanol production, whereas lignin is considered as a waste and normally is burned to
and increases of the yields of the fermentable sugars. Various chemicals and reagents have
generate energy. In the syngas platform, all three components are used for ethanol pro-
been used for biomass pretreatment process research and development. Some of these
duction. The key steps of the sugar platform and the syngas platform are shown in Figure
chemicals and reagents together with their corresponding substrates are summarized in
3 and Figure 4, respectively.
Table 2.

Lignocellulosic
biomass

Chemical Hydrolysis
Grinding (concentrated H2SO4,
supercritical CO2)

Pretreatment

Enzymatic Fermentable
Hydrolysis Sugars

Fermentation Ethanol

Figure 3. The sugar platform for ethanol production from biomass.

Figure 3. The sugar platform for ethanol production from biomass.
Fermentation 2021, 7, 314 8 of 18
Fermentation 2021, 7, x FOR PEER REVIEW 8 of 19

Lignocellulosic
biomass

Grinding

Gasification

Synthesis Fermentation Ethanol

Gas (Syngas)

Chemical
Catalysis

Figure 4. The syngas platform for ethanol production from biomass.

Figure 4. The syngas platform for ethanol production from biomass.

Hydrolysis
Among the of manycellulose and hemicellulose
pretreatment processes thereto generate
has notfermentable sugars for
been a clear winner. The ethanol
use of
fermentation can be performed with either chemicals or enzymes.
superheated steam, however, seemed to be a favorite. It has been used in the commercial The chemicals used for
the hydrolysis of cellulose and hemicellulose include concentrated
Inbicon process for pretreatment of wheat straw [46]) and more recently in the Clariant (>70%) H 2SO 4 , which
is used in ethanol
cellulosic the Arkenolplantprocess [21]Romania
in Podari, and the [47].
Biosulfurol process [22],
The pretreatment supercritical
processes havefluid,
been
which
reviewedis used in the Renmatix
thoroughly in severalprocess
reviews[23], and γ-valerolactone
[8,48–50]. The fermentable (GVL), which
sugars is used in
obtained
the GlucanBio
enzymatic processof[24].
hydrolysis cellulose and hemicellulose consists mostly of glucose and xylose,
plus When concentrated
arabinose, galactose,Hand2SO4mannose
was usedin forsignificantly
hydrolysis, smaller
the sugar productSince
amounts. was athe mixture
yeast
of
S. glucose
cerevisiaeand xylose
cannot whereas in
metabolize the other two
five-carbon cases,
sugars twoasseparate
such streams thatengineered
xylose, genetically contained
mostly
strains glucose
capableand xylose, respectively,
of utilizing both glucose were
andformed.
xylose forTheethanol
fractionation of the two
production havecarbo-
been
hydrate
developed fractions would allow
for commercial the use of
applications the C6 sugar stream for ethanol production by
[51].
the native
VariousS. ceresiae
types ofstrains and the
commercial C5 sugar
gasifiers arestream
suitable forforproduction of higher-value
biomass gasification. The co-
im-
purities and
products suchinhibitory
as xylitol compounds
and astaxanthinin the syngas produced have to be removed before
[25].
the syngas
Due tocanthebe used
rigid for ethanol
structure production
of biomass, whichin either
impedesthe microbial
enzymaticorattacks,
catalytic
a process.
process
Biomass
called gasification is
pretreatment has been reviewed
required in detail
before the by Ciliberti
cellulose et al. [52]. The
and hemicellulose microorganisms
components of bi-
that have been considered as most promising candidates for commercial
omass can be effectively hydrolyzed by enzymes. In the pretreatment process the rigid syngas ethanol
fermentation
structure includeisClostridium
of biomass opened upljungdahlii,
and amorphous Clostridium
regions ragsdalei, Clostridium
are created. carboxydivo-
The results of the
rans, Clostridium
pretreatment processcoskatii, Clostridium autoethanogenum,
are improvements of the rates of the Alkalibaculum
andsubsequent bacchi. Some
enzymatic hydrol-of
theseand
ysis species only of
increases produce ethanol
the yields and
of the acetic acid,sugars.
fermentable whereas others chemicals
Various can produce andadditional
reagents
products,
have beenwhich
used include butyric
for biomass acid, butanol,
pretreatment and 2, research
process 3-butanediol andas mentioned previously.
development. Some of
One of the key issues of ethanol production from biomass
these chemicals and reagents together with their corresponding substrates are summa-via fermentation, either in
the sugar platform
rized in Table 2. or the syngas platform, is the low achievable final ethanol concentration.
Since it is most likely that the ethanol produced will be recovered by distillation, which is a
very energy-intensive process, the minimum acceptable ethanol concentration in the input
to the distillation unit is 50 g/L [8].
Fermentation 2021, 7, 314 9 of 18

Table 2. Summary of biomass pretreatment chemicals and agents [26–45].

Chemicals/Reagents Biomass Type References

Dilute acids
H2 SO4 Doulas fir chip Nguyen et al. 1998 [26]
HCl Sorghum straw Herrera et al. 2003 [27]
H3 PO4 Sugarcane bagasse Geddes et al. 2010 [28]
HNO3 Corn stover Zhang et al. 2011 [29]
Alkalines
NaOH Rice straw Castro et al. 2017 [30]
KOH Hybrid poplar Gupta and Lee 2010 [31]
NH4 OH Corn stover Le et al. 2021 [32]
Anhydrous NH3 Hybrid poplar Balan et al. 2009 [33]
Ca(OH)2 (Lime) Corn stover Kaar and Holtzapple 2000 [34]
Basic Salts
Green liquor Sugarcane bagasse Zhou et al. 2016 [35]
Na2 CO3/ Na2 S Sweet sorghum bagasse Pham et al. 2018 [36]
Na2 CO3 NaOH Sweet sorghum bagasse Nghiem and Toht 2020 [37]
Na3 PO4 Corn stover Qing et al. 2016 [38]
Water and Steam Spruce wood chips
High pressure team Pielhop et al. 2016 [39]
Liquid hot water Poplar Li et al. 2017 [40]
Organic solvents
Methanol Corn stover Qing et al. 2017 [41]
Ethanol (with/without acid catalyst) Various Zhang et al. 2016 [42]
Ethylen glycol Palm tree residues Ariols et al. 2009 [43]
Glycerol Rice straw Trinh et al. 2016 [44]
γ-velerolactone Hard wood Shuai et al. 2016 [45]

3. Global Ethanol Production

It is estimated that currently 60% ethanol is produced from corn, 25% from sugar
cane, 3% from wheat, 2% from molasses, and the rest from other grains, cassava and sugar
beets. The annual world fuel ethanol production in the last three years together with the
production by the individual countries that contributed to at least 1% of the total production
are summarized in Table 3. As indicated in Table 3, the top five ethanol producers before
the COVID-19 pandemic were the United States, Brazil, the European Union (EU), China,
and Canada. In 2020, India produced slightly more ethanol than Canada but the ranking of
the top four ethanol producers did not change.

Table 3. Annual world fuel ethanol production (Mil. L) (Source: Renewable Fuels Association
analysis of public and private data sources [53].

Region 2018 2019 2020 % of 2020 World Production

United States 16,091 15,778 13,926 53%
Brazil 7990 8590 7930 30%
European Union (EU) 1450 1370 1250 5%
China 770 1000 880 3%
Canada 460 520 428 2%
India 430 510 515 2%
Thailand 390 430 400 2%
Argentina 290 280 230 1%
Rest of the world 529 522 500 2%
Total 28,400 29,000 26,059
Note: The EU in this review refers to the union of 28 members, i.e., before the exit of the United Kingdom.

The number of production plants, plant type, feedstocks, and co-products of the
ethanol industry in the top five ethanol-producing countries/regions are summarized in
Table 4. In this table, all the data are for 2020. The case of the United States needs an
explanation. Several corn dry-grind ethanol plants in the United States used a bolt-on
Fermentation 2021, 7, 314 10 of 18

process such as the D3MAX or QCCP process discussed previously to convert the corn
fibers, i.e., a biomass feedstock, and the non-converted corn starch to additional ethanol.
The quantities of ethanol produced from each feedstock in these processes could not be
determined separately. Therefore, all the ethanol produced in these plants are listed under
corn ethanol production. In addition, some plants occasionally used small amounts of
sorghum grains to mix with corn. The quantities of sorghum grains used were not known.
Therefore, all the starch-based feedstock is listed under corn.

Table 4. The number of ethanol plants, plant type, feedstocks, and co-products in the top five
ethanol-producing countries/regions [54–58].

Country/Region USA Brazil EU China India

Number of 1st-generation plants 208 360 57 18 220
Nameplate capacity (Bil L) 65.8 42.8 8.15 6.58 3.50
Capacity used (%) 80 67 58 49 85
Number of 2nd-generation plants 3 3 3 1 0
Nameplate capacity (Bil L) 0.21 0.13 0.09 0.07
Capacity used (%) n/a 25 28 0
Feedstocks (1000 MT)
Corn 123,465 5995 6350 7100 0
Other grains 0 0 4300 900 0
Cassava 0 0 0 1000 0
Sugarcane 0 326,630 0 0 0
Sugar beets 0 0 7450 0 0
Molasses 0 0 0 0 6407
Biomass 0 0.178 200 200 0
Co-products (1000 MT)
Bagasse 0 120,077 0 0 118,374
Distillers grains 29,437 1876 3332 2348 0
Corn gluten feed 3087 0 0 0 0
Corn gluten meal 605 0 0 0 0
Corn Oil 27.9 108.0 184 0 0
Note: n/a: Not available.

Ethanol production from the individual nations with significant annual ethanol out-
puts are reviewed in the following sections.

3.1. North America

3.1.1. The United States
In the United States, ethanol produced from corn reached 6.5 billion gallons (24.6 billion L)
in 2007 and in 2012 this total doubled to approximately 13.2 billion gallons (50.0 billion L).
In 2018 the United States exported a total of 6.5 billion L of ethanol, expanding the share
of world exports of ethanol to 61%. Most ethanol produced in the United States is from
starch-based crops by dry or wet-mill processing technologies. In 2020, there were 208 ethanol
production plants in the United States with a total installed capacity of 17.44 billion gallons
per year (66.0 billion L per year). The total production in 2020 was 13.8 billion gallons
(52.2 billion L), which represented a 12.7% decrease from the production total of 15.8 billion
gallons (59.8 billion L) in the previous year [11,59]. The decrease in ethanol production
was mainly caused by the reduction in transportation fuel demand, which was the direct
consequence of the COVID-19 pandemic. Whereas a number of plants were shut down, some
plants modified the production process to produce an ethanol product which is suitable for
use in hand sanitizers to cope with the economic downturn. It is expected that production
of ethanol will go back up in 2021. The ethanol co-products generated in 2020 included
29.4 million metric tons (MT) distillers grains, 3.1 million MT corn gluten feed, 0.6 million
MT corn gluten meal, 1.5 million MT corn oil and 2.1 million MT captured CO2 . Corn
was the major feedstock, which accounted for 93.2% of the total ethanol production in 2020.
Fermentation 2021, 7, 314 11 of 18

Other feedstocks included sorghum, corn fiber, waste sugars, waste starch and lignocellulosic
biomass. The interest in biomass as a potential feedstock for ethanol production continued
to be strong. Despite previous failures by large corporations such as Poet and DuPont, New
Energy Blue announced in July 2021 the plan to construct a full-scale ethanol biorefinery,
which would consume 250,000 MT per year agricultural residues generated locally for ethanol
production. The proposed plant would use the Inbicon process, which would also produce
a lignin co-product for use as a solid biofuel and a natural binder and possibly xylitol as a
specialty chemical with potential food applications [60,61].

3.1.2. Canada
Canada is a net importer of ethanol since the volume consumed in Canada generally
exceeded the ethanol this country produced. In 1980, Canadian produced merely 8000 L
but in 2010 the annual ethanol production of Canada reached 1.9 billion L. In 2019, ethanol
consumption in Canada reached 3.33 billion L compared to 2 billion L in production.
However, ethanol production in Canada has grown in recent years largely due to changes
in feedstocks and increased capacity at existing ethanol facilities in Canada.
In 2020, Canada was ranked as the sixth largest ethanol producer in the world. The
ethanol production in Canada represented 1.6% of the total global production. The two
major feedstocks used for ethanol production were corn and wheat, which contributed
1534.3 million L and 360.7 million L, respectively [59]. Winter barley has been considered
as a potential feedstock for ethanol production. However, this grain has not been used in
any commercial ethanol production plant.

3.2. South America

3.2.1. Brazil
Brazil continued to be the second largest ethanol producer in the world. In 2019, Brazil
ethanol production including anhydrous and hydrous ethanol amounted to more than
35.3 million M3 , an increase of nearly seven percent in comparison to a year of 2018. This
is the highest volume of fuel ethanol that Brazil has produced in the decade. In Brazil,
sugarcane bagasse is commonly used as boiler fuel to produce energy to supply sugar mills.
This is practiced to minimize the energy costs and also as an alternative to the utilization of
the biomass left over.
In 2020, the ethanol production in Brazil represented 26.7% of the total global ethanol
production [53]. There were 360 first-generation (sugar cane, corn) ethanol plants in Brazil
in 2020 with a total nameplate capacity of 42,800 million L. However, only 67% of the
capacity was used. There were three plants that used lignocellulosic feedstock with a
total nameplate capacity of 127 million L. In 2020, these second-generation ethanol plants
only operated at 25% capacity. The feedstocks included sugar cane (326.6 million MT),
corn (6.0 million MT) and bagasse (178,000 MT). The total ethanol production in 2020
was 31.35 billion L, which included 32 million L produced from lignocellulosic feedstock
(bagasse). The 2020 ethanol production was about 16% lower compared to the previous
year (37.38 billion L). The decrease in ethanol production in 2020 was mainly caused by the
diversion of sugar cane juice toward more sugar production in sugar-ethanol plants. The
co-products included 120.1 million MT bagasse, 1.88 million MT DDGS and 108,000 MT
corn oil. The traditional feedstock for first-generation ethanol production in Brazil is
sugar cane. However, corn has become an important feedstock and its use for ethanol
production has steadily gained ground. In 2020, 2.5 billion L ethanol was produced from
corn, which was 1.17 million L higher than the previous year. The Corn Ethanol National
Union (UNEM) predicted that corn ethanol production would reach 8 billion L by 2028.
There currently are 11 corn ethanol plants, which include nine full-plant types (corn only)
and 2 flex-plant types (corn and sugarcane). Two full-plants and one flex-plant are currently
under construction [55].
Fermentation 2021, 7, 314 12 of 18

3.2.2. Argentina
In 2020, Argentina was ranked 8th in the world with a contribution of 1.0% toward the
total global ethanol production [53]. There were 22 ethanol plants with a total nameplate
capacity of 1580 million L per year. The ethanol plants, however, were operated only
at 55.1% capacity. Corn and molasses were used as feedstocks at 1.09 million MT and
1.70 million MT, respectively. The total ethanol production was 870 million liters, which
was significantly lower than the total production in the previous year (1073 million L). The
DDGS co-product was also produced at 345,000 MT [62].

3.2.3. Colombia
Colombia was ranked 13th in the world in 2020 with a contribution of 0.44% toward
the total global ethanol production [60]. The country had six ethanol plants, which used
sugar cane as the only feedstock. The total nameplate capacity of the plants was 540 million
liters. These plants were run at only 73.1% capacity in 2020. The total ethanol production in
2020 was 395 million liters with 1.34 million MT bagasse co-product. Five of the six ethanol
plants were directly linked to the sugar production plants. The bagasse obtained after juice
extraction was used to generate energy for internal use in these plants. Most ethanol plants
in Colombia were energy self-sufficient and even generated surplus energy to sell to the
national grid [63].

3.3. Europe
The European Union
The total ethanol production in the EU in 2020 was 4.8% of the total global produc-
tion [53]. The major feedstocks used for ethanol production were sugar beets (7.45 million
MT), corn (6.35 million MT), and wheat (2.64 million MT). Other first-generation feedstocks
were triticale (1.04 million MT), rye (520,000 MT) and barley (450,000 million MT). Wheat
was predominantly used in Belgium, Germany, France and the UK. Corn was the preferred
feedstock in Hungary, where corn was abundantly available, and in the Netherlands, Spain,
and the UK, where the majority of corn came from the Ukraine to provide the feedstock for
ethanol plants, which are located near seaports. The inland ethanol plants in Spain used a
combination of corn and barley as the feedstock. Sugar beets and their derivatives were
used for ethanol production in France, Germany, the UK, the Czech Republic, Belgium
and Austria. The usage of lignocellulosic feedstocks in 2020 was doubled to 200,000 MT
compared to the previous year. There were 57 first-generation ethanol plants with a total
nameplate capacity of 8.15 billion L, which were operated at 58% capacity. There were also
3 lignocellulosic ethanol plants with a total nameplate capacity of 90 million L. However,
these second-generation ethanol plants were operated at only 28% capacity. Five other
lignocellulosic ethanol plants were under construction in Finland (sawdust, 10 million L
per year), Italy (biomass, 28 million L per year), Austria (wood sugars, 30 million L per
year), Romania (wheat straw, 65 million L per year) and Bulgaria (corn stover, 50 million L
per year). All of these plants are expected to be in operation soon. The ethanol production
in the EU in 2020 was 5.47 billion L. Due to the COVID-19 pandemic, ethanol consumption
in the EU has decreased by 10.1%. However, this number was still slightly lower than the
13.0% decrease in gasoline consumption. According to the 2021 European Union Biofuels
Annual Report, the co-products included 3.33 million MT DDGS and 188,000 MT corn
oil [56].

3.4. Asia and Rest of the World

3.4.1. Vietnam
Vietnam is an agriculture-based country and has abundant natural resources for
renewable power development. However, the pace of renewable energy development
including ethanol production is not yet rapid due to barriers such as the small size of
the country’s economy, the lack of financial capacity, advanced technologies and human
Fermentation 2021, 7, 314 13 of 18

resources. Institutional barriers such as market-controlled mechanisms and unstable

supporting policies also limit the development of renewable power sectors [64].
In terms of ethanol development, due to the economic development, Vietnam has
consumed lots of gasoline in recent years. The United States Department of Agriculture
(USDA) estimated gasoline consumption of Vietnam in recent years has grown approxi-
mately 4–5 percent per year. In the first quarter of 2020, Vietnam spent about $2.5 billion on
importing crude oil and petroleum products, including ethanol, in addition to its produc-
tion [65]. Since 2007 ethanol supporting regulations have gradually increased, however, the
commercialization and sales of five percent ethanol blended gasoline (E5) was pushed back
to 2018 due to a lack of understanding of the environmental benefits of ethanol blended
gasoline by consumers, together with some persistent rumors and myths that usage of
ethanol might harm vehicle engines.

3.4.2. Korea
Korea is the 7th ranked CO2 emitter in the world and similar to other countries is
concerned about its high CO2 emissions and its dependence on imported crude oil. All of
the oil Korea consumes is imported from foreign countries. South Korea is a main importer
of United States ethanol, with a total volume of 263.84 million L (24.7 million bushels in
corn equivalent) in 2017–2018. In the mid-1990s, production of ethanol for fuel use in Korea
using imported cassava as a feedstock was initiated. The Korea Ministry of Knowledge
Economy (KMKE) announced an action plan to increase the use of biofuels in the transport
sector from 0.2 billion L in 2008 to 5 billion L by 2030 [66].

3.4.3. China
From 2004 to 2016, China produced ethanol with an average annual increase in
production rate of 16.8 percent. In 2017, China had an ethanol production of 2.8 million MT.
In 2018, China produced 6.6 million MT, making it the fourth-largest ethanol producing
country/region in the world, after the United States, Brazil, and the EU. China set a policy
that gasoline supplies across the country were required to be blended with ethanol by 2020.
According to a news source, China aimed to have 15 million MT by 2020, which is almost
triple its current ethanol production capacity, in order to keep up with growing demands
for cleaner fuels [57]. The target would exceed the estimated domestic production capacity
of China and the country would need to import ethanol from foreign countries such as the
United States and Brazil. Corn is China’s main feedstock, which is currently accounting for
64 percent of total output for China’s ethanol production [67].

3.4.4. India
India ranked 6th among the leading ethanol producers in the world. In 2020, India
still remained one of the biggest importers of the United States ethanol, with a market
share of 99 percent. With Modi’s “self-resilient” strategies, India sets its ambitious goal of
E-20 by 2025 while retaining its immediate goal of E-10 by 2022. India has a total installed
ethanol capacity of 5 billion L, of which molasses-based distilleries constitute 4.2 billion L,
or 85 percent of the overall production capacity, while grain-based distilleries constitute
750 million L (equivalent to 15 percent).
Formerly, ethanol in India could only be produced from molasses or sugar juice.
However, India’s existing ethanol is produced from a variety of feedstocks such as cereals
(rice, wheat, barley, maize, and sorghum). In 2020, an estimated 2.98 billion L of ethanol was
produced from molasses. In 2021, India’s ethanol production was forecast at 3.17 billion L,
7% above 2020 due to surplus sugarcane production, and 2021 average ethanol blending
rate in gasoline of India was estimated at 7.5 percent, due to accelerated government efforts
to divert more feedstock toward ethanol [68].
Fermentation 2021, 7, 314 14 of 18

3.4.5. Thailand
Thailand is the 7th largest ethanol producer in the world. Thailand consumed
1500 million L of ethanol in 2020. Thailand’s 20-year Alternative Energy Development
Plan (AEDP) for 2018–2037 targeted 2.0 billion L for ethanol in 2021 and 2.7 billion L for
ethanol consumption in 2037. Molasses is the primary feedstock for ethanol production
in Thailand. Molasses supplies in Thailand have been tight due to reduced sugarcane
production for two consecutive years. Ethanol demand is primarily expected to be fulfilled
by cassava-based ethanol in Thailand [69].

3.4.6. Australia
Australia’s ethanol consumption is forecast to remain stable in 2020 at only 1.4%
of gasoline consumption. Ethanol in Australia is primarily produced as a biofuel for
passenger and commercial vehicles and also for alcoholic beverage, industrial chemicals,
and solvents used in pharmaceutical and cosmetic applications. In spite of large feedstock
availability, Australia’s ethanol production volumes remain small because there is no
nationwide fuel ethanol program. In Australia only two states, which are New South
Wales and Queensland, have mandates with a fuel ethanol program, and have the highest
consumption of ethanol blended fuel.
In New South Wales, Manildra is the largest ethanol producer with a capacity of
over 300 million L. In this plant wheat starch is processed through an integrated process
which separates the gluten and processes the remaining starch into a range of food and
industrial-grade starches, glucose syrups, and ethanol products. In Queensland, a plant at
Dalby producing ethanol from starch-based feedstock is operated by United Petroleum.
This biorefinery with a capacity of 80 million L fuel-grade ethanol is located in a sorghum
growing region in the Darling Downs and processes up to 0.2 million MT of sorghum grain
a year from local growers. At full capacity, the biorefinery can also produce 830,000 MT of
wet distillers grain, which is used for animal feed supplements [70].

3.4.7. Zimbabwe and Other African countries

Zimbabwe has adopted ethanol petrol blending regulations. Two ethanol plants in
Zimbabwe, namely Triangle Sugar and Hippo Valley, were installed. In 2010, the Triangle
plant resumed ethanol production after refurbishing its ethanol plant with a capacity
of 27 million L per year. The government of Zimbabwe also initiated the Chisumbanje
sugar/bioethanol project with an aim of using 10,000 hectares of sugarcane for this project.
In March 2012, owners of this project halted production because the plant was running out
of storage space. In Zimbabwe, some oil companies were only selling E10 at a few of their
filling stations [71].
In other African countries, lead additives are still heavily used in gasoline and where
sugarcane production cost is high, ethanol can be a cheap source of octane in gasoline. It is
estimated that in Africa, to replace all the lead used in African gasoline, this would require
Africa to produce about 20% of the amount of ethanol currently produced in Brazil, and
would require the shift of some sugar production to ethanol production. African countries
that could replace lead with ethanol using primarily their by-product molasses production
include Zimbabwe, Kenya, Egypt, Zaire, Zambia, Sudan, Swaziland, and Mauritius at a
more modest scale [72].

4. Discussion
Over the last several decades, ethanol has emerged to become an important trans-
portation liquid fuel. The corn ethanol in the United States and the sugarcane ethanol in
Brazil have gained their position as a mature industry. The strong interest in ethanol from
sustainable sources has led to the development of process technology for production of
ethanol from biomass. The research in this area has led to many technological achieve-
ments and changes of the biobased product industry. One example is the development
of biomass pretreatment processes, which have been improved to shift the focus on the
Fermentation 2021, 7, 314 15 of 18

cellulose fraction in biomass to include hemicellulose utilization. In the early developments

of pretreatment processes, lignin was considered as a waste, which was to be burned to
provide the thermal energy required in a biomass ethanol plant. The research on processes
for conversion of lignin to high-value products has changed this view and lignin now is
considered as a co-product of biomass ethanol production. Another example is the devel-
opments of much more efficient of commercial cellulase and xylanase products. Despite
several failures of biomass ethanol production plants, others continued to be constructed
with improved technical knowledge and better production strategies. It is anticipated that
the second-generation ethanol industry will eventually emerge and establish itself similar
to the first-generation ethanol based on starch-based and sugar-based feedstocks.

5. Conclusions and Global Trend Forecast

As discussed previously, corn ethanol in the United States and sugarcane ethanol in
Brazil have been in commercial practice for many years and hence there is little room for
research to significantly improve the technology. However, in the area of biomass ethanol,
there are still opportunities for research to improve the technology and to bring it closer to
commercial success. One area that needs further research is the development of high-value
co-products that can be produced from the C5 sugars and the processes for their production.
Examples of these products are xylitol and astaxanthin, among others. Another area that
still offers many research opportunities is lignin utilization. Similar to the case of the C5
sugars, lignin can be used as a feedstock for production of high-value coproducts and
development of processes for their production still requires significant research efforts.
COVID-19 has hit the world and the production industries worldwide have been
seriously affected. As a result, ethanol output worldwide dropped in 2020. Other areas are
being hit as well but their lower reliance on the fuel ethanol markets make the negative
impact of the pandemic have been relatively less. When the pandemic is over and the pro-
duction industries return to normalcy, it is expected that the rebound will come, however,
the production might not be as strong as before the lock-down period worldwide. During
the pandemic, even after the restrictions are eased, demand on ethanol for hand-sanitizer
manufacturing is high. In developing countries, stronger emissions regulations and policies
help to encourage the production potential for biofuels to offset CO2 emissions. With an
eye toward the future, worldwide ethanol production is expected to increase again.

Author Contributions: Conceptualization, T.-D.H. and N.N.; methodology, T.-D.H. and N.N.; valida-
tion, T.-D.H. and N.N.; formal analysis, T.-D.H. and N.N.; investigation, T.-D.H. and N.N.; resources,
T.-D.H. and N.N.; data curation, T.-D.H. and N.N.; writing—original draft preparation, T.-D.H.
and N.N.; writing—review and editing, T.-D.H. and N.N.; visualization, T.-D.H. and N.N.; project
administration, T.-D.H. and N.N. All authors have read and agreed to the published version of the
manuscript.
Funding: This research received no external funding.
Institutional Review Board Statement: Not applicable.
Informed Consent Statement: Not applicable.
Data Availability Statement: Not applicable.
Conflicts of Interest: The authors declare no conflict of interest.

References
1. Global Ethanol Production by Country or Region. Available online: https://afdc.energy.gov/data/10331/ (accessed on 19
September 2021).
2. The Future of Biofuels: A Global Perspective. Available online: https://www.ers.usda.gov/amber-waves/2007/november/the-
future-of-biofuels-a-global-perspective/ (accessed on 19 September 2021).
3. Halde, P.; Azad, K.; Shah, S.; Sarker, E. Prospects and technological advancement of cellulosic bioethanol ecofuel production. Adv.
Eco-Fuels Sustain. Environ. 2019, 211–236. [CrossRef]
Fermentation 2021, 7, 314 16 of 18

4. U.S. Energy Information Administration. Biofuels Explained, Ethanol. Available online: https://www.eia.gov/energyexplained/
biofuels/ethanol.php (accessed on 1 December 2021).
5. Ruangrit, K.; Chaipoot, S.; Phongphisutthinant, R.; Kamopas, W.; Jeerapan, I.; Pekkoh, J.; Srinuanpan, S. Environmental-friendly
pretreatment and process optimization of macroalgal biomass for effective ethanol production as an alternative fuel using
Saccharomyces cerevisiae. Biocatal. Agric. Biotechnol. 2021, 31, 101919. [CrossRef]
6. Environmetnal and Energy Study Institute’s Report on Bioenery (Biofuels and Biomass). Available online: https://www.eesi.
org/topics/bioenergy-biofuels-biomass/description (accessed on 1 December 2021).
7. Brazil Biofuels Annual Report 2016, Global Agricultural Information Network; GAIN Report Number BR16009; Foreign Agricultural
Service, U.S. Department of Agriculture: Washington, DC, USA, 2016.
8. Drapcho, C.; Nghiem, N.; Walker, T. Ethanol Production. In Biofuels Engineering Process Technology, 2nd ed.; McGraw-Hill: New
York, NY, USA, 2020; pp. 117–271.
9. Andrietta, S.R.; Maugeri, F. Optimum design of a continuous fermentation unit of an industrial plant for alcohol production. In
Advances in Bioprocess Engineering; Galindo, E., Ramirez, O.T., Eds.; Kluwer Academic Publishers: Dordrecht, The Netherlands,
1994; pp. 47–52.
10. España-Gamboa, E.; Mijangos-Cortes, J.; Barahona-Perez, L.; Dominguez-Maldonado, J.; Hernández-Zarate, G.; Alzate-Gaviria, L.
Vinasses: Characterization and treatments. Waste Manag. Res. 2011, 29, 1235–1250. [CrossRef]
11. Renewable Fuels Association. 2021 Ethanol Industry Outlook. Available online: https://ethanolrfa.org/wp-content/uploads/20
21/02/RFA_Outlook_2021_fin_low.pdf/ (accessed on 22 September 2021).
12. Enogen Corn for Ethanol. Available online: https://www.syngenta-us.com/corn/enogen/ (accessed on 22 September 2021).
13. TransFerm® Yield+. Available online: https://www.lallemandbds.com/products/transferm-yield/ (accessed on 22 September
2021).
14. Lallemand Launches New Convergence Yeast Enzyme Platform: Could Save Ethanol Producers $1M/Year in Replacing All
External Fermentation Enzymes. Available online: https://www.biofuelsdigest.com/bdigest/2020/09/14/lallemand-launches-
new-convergence-yeast-enzyme-platform-could-save-ethanol-producers-1m-year-in-replacing-all-external-fermentation-
enzymes/ (accessed on 22 September 2021).
15. Novozymes Adds Yeast to the Bioenergy Business. Available online: https://www.novozymes.com/en/news/news-archive/20
18/02/novozymes-adds-yeast-to-bioenergy-business/ (accessed on 22 September 2021).
16. Yeast for Fermentation in Bioenergy. Available online: https://biosolutions.novozymes.com/en/bioenergy/fermentation/
(accessed on 22 September 2021).
17. D3MAX Technology. Available online: https://www.d3maxllc.com/technology/ (accessed on 22 September 2021).
18. American Coalition for Ethanol. Available online: https://ethanol.org/DelayneJohnsonQuietIngenuity.pdf/ (accessed on 22
September 2021).
19. California Certifies 2 Plants for Corn Kernel Fiber Ethanol. Available online: http://ethanolproducer.com/articles/16203/
california-certifies-2-plants-for-corn-kernel-fiber-ethanol/ (accessed on 22 September 2021).
20. Intellulose: An Innovative Approach to Your Plant’s Profitability. Available online: https://distillersgrains.org/wp-content/
uploads/2019/05/7-Kacmar-Intellulose.pdf/ (accessed on 22 September 2021).
21. Farone, W.A.; Cuzens, J.E. Method of Producing Sugars Using Strong Acid Hydrolysis of Cellulosic and Hemicellulosic Materials.
U.S. Patent 5,562,777, 8 October 1996.
22. Groenestijn, J.W.; Hazewinkel, J.H.O.; Bakker, R.R. Pre-treatment of lingo-cellulose with biological acid recycling (the Biosulfurol
process). Int. Sugar J. 2008, 110, 689–692.
23. Kilambi, S.; Kadam, K.L. Production of Fermentable Sugars and Lignin from Biomass Using Supercritical Fluids. U.S. Patent
2013/0239954, 21 August 2018.
24. Dumesic, J.A.; Alonso, D.M.; Gürbüz, E.I.; Wettstein, S.G. Production of Levulinic Acid, Furfural, and Gamma-Valerolactone from
C5 and C6 Carbohydrates in mono- and Biphasic Systems Using Gamma-Valerolactone as a Solvent. U.S. Patent 8,399,688, 29
November 2012.
25. Zhang, X.; Nghiem, N.P. Pretreatment and Fractionation of Wheat Straw for Production of Fuel Ethanol and Value-added
Co-products in a Biorefinery. AIMS Bioeng. 2014, 1, 40–52. [CrossRef]
26. Nguyen, Q.A.; Tucker, M.P.; Boynton, B.L.; Keller, F.A.; Schell, D. Dilute acid pretreatment of softwoods. Appl. Biochem. Biotechnol.
1998, 70–72, 77–87. [CrossRef]
27. Herrera, A.; Téllez-Luis, S.J.; Ramírez, J.A.; Vázquez, M. Production of xylose from sorghum straw using hydrochloric acid. J.
Cereal Sci. 2003, 37, 267–274. [CrossRef]
28. Geddes, C.C.; Roslander, J.J.P.C.; Zacchi, G.; Mullinnix, M.T.; Shanmugam, K.T.; Ingram, L.O. Optimizing the saccharification of
sugar cane bagasse using dilute phosphoric acid followed by fungal cellulases. Bioresour. Technol. 2010, 101, 1851–1857. [CrossRef]
[PubMed]
29. Zhang, R.; Lu, X.; Sun, Y.; Wang, X.; Zhang, S. Modeling and optimization of dilute nitric acid hydrolysis on corn stover. J. Chem.
Technol. Biotechnol. 2011, 86, 306–314. [CrossRef]
30. Castro, R.C.d.; Fonseca, B.G.; Santos, H.T.L.; Ferreira, I.S.; Mussatto, S.I.; Roberto, I.C. Alkaline deacetylation as a strategy to
improve sugars recovery and ethanol production from rice straw hemicellulose and cellulose. Ind. Crops Prod. 2017, 106, 65–73.
[CrossRef]
Fermentation 2021, 7, 314 17 of 18

31. Sharma, R.; Palled, V.; Sharma-Shivappa, R.R.; Osborne, J. Potential of potassium hydroxide pretreatment of switchgrass for
fermentable sugar production. Appl. Biochem. Biotechnol. 2013, 169, 761–772. [CrossRef]
32. Le, T.D.T.; Truong, V.P.N.; Ngo, M.T.T.; Kim, T.H.; Oh, K.K. Pretreatment of Corn Stover Using an Extremely Low-Liquid
Ammonia (ELLA) Method for the Effective Utilization of Sugars in Simultaneous Saccharification and Fermentation (SSF) of
Ethanol. Fermentation 2021, 7, 191. [CrossRef]
33. Balan, V.; Sousa, L.d.; Chundawat, S.P.S.; Marshall, D.; Sharma, L.N.; Chambliss, C.K.; Dale, B.E. Enzymatic digestibility and
pretreatment degradation products of AFEX-treated hardwoods (Populus nigra). Biotechnol. Prog. 2009, 25, 365–375. [CrossRef]
34. Kaar, W.E.; Holtzapple, M.T. Using lime pretreatment to facilitate the enzymatic hydrolysis of corn stover. Biomass Bioenergy 2000,
18, 189–199. [CrossRef]
35. Zhou, Z.; Xue, W.; Lei, F.; Cheng, Y.; Jiang, J.; Sun, D. Kraft GL-ethanol pretreatment on sugarcane bagasse for effective enzymatic
hydrolysis. Ind. Crops Prod. 2016, 90, 100–109. [CrossRef]
36. Pham, H.T.T.; Nghiem, N.P.; Kim, T.H. Near theoretical saccharification of sweet sorghum bagasse using simulated green liquor
pretreatment and enzymatic hydrolysis. Energy 2018, 157, 894–903. [CrossRef]
37. Nghiem, N.P.; Toht, M. Pretreatment of Sweet Sorghum Bagasse for Ethanol Production Using Na2 CO3 Obtained by NaOH
Absorption of CO2 Generated in Sweet Sorghum Juice Ethanol Fermentation. Fermentation 2019, 5, 91. [CrossRef]
38. Qing, Q.; Zhou, L.; Guo, Q.; Huang, M.; He, Y.; Wang, L.; Zhang, Y. A combined sodium phosphate and sodium sulfide
pretreatment for enhanced enzymatic digestibility and delignification of corn stover. Bioresour. Technol. 2016, 218, 209–216.
[CrossRef]
39. Pielhop, T.; Amgarten, J.; von Rohr, P.R.; Studer, M.H. Steam explosion pretreatment of softwood: The effect of the explosive
decompression on enzymatic digestibility. Biotechnol. Biofuels 2016, 9, 152. [CrossRef]
40. Li, M.; Cao, S.; Meng, X.; Studer, M.; Wyman, C.E.; Ragauskas, A.J.; Pu, Y. The effect of liquid hot water pretreatment on the
chemical–structural alteration and the reduced recalcitrance in poplar. Biotechnol Biofuels 2017, 10, 237. [CrossRef] [PubMed]
41. Qing, Q.; Zhou, L.; Guo, Q.; Gao, X.; Zhang, Y.; He, Y.; Zhang, Y. Mild alkaline presoaking and organosolv pretreatment of corn
stover and their impacts on corn stover composition, structure, and digestibility. Bioresour. Technol. 2017, 233, 284–290. [CrossRef]
[PubMed]
42. Zhang, K.; Pei, Z.; Wang, D. Organic solvent pretreatment of lignocellulosic biomass for biofuels and biochemical: A review.
Bioresour. Technol. 2016, 199, 21–33. [CrossRef] [PubMed]
43. Alriols, M.G.; Tejado, A.; Blanco, M.; Mondragon, I.; Labidi, J. Agricultural palm oil tree residues as raw material for cellulose,
lignin and hemicelluloses production by ethylene glycol pulping process. Chem. Eng. J. 2009, 148, 106–114. [CrossRef]
44. Trinh, L.T.P.; Lee, J.; Lee, H. Acidified glycerol pretreatment for enhanced ethanol production from rice straw. Biomass Bioenergy
2016, 94, 39–45. [CrossRef]
45. Shuai, L.; Questell-Santiago, Y.M.; Luterbacher, J.S. A mild biomass pretreatment using γ-valerolactone for concentrated sugar
production. Green Chem. 2016, 18, 937–943. [CrossRef]
46. Larsen, J.; Haven, M.Ø.; Thirup, L. Inbicon makes lignocellulosic ethanol a commercial reality. Biomass Bioenergy 2012, 46, 36–45.
[CrossRef]
47. Kennedy, H.T. Big Cellulosic: Inside the Clariant Cellulosic Ethanol at Podari. Available online: https://www.biofuelsdigest.
com/bdigest/2021/10/20/big-cellulosic-inside-the-clariant-cellulosic-ethanol-at-podari-romania/ (accessed on 22 November
2021).
48. Galbe, M.; Wallberg, O. Pretreatment for biorefineries: A review of common methods for efficient utilisation of lignocellulosic
materials. Biotechnol. Biofuels 2019, 12, 294. [CrossRef]
49. Anukam, A.; Berghel, J. Biomass Pretreatment and Characterization: A Review, Open-Access Pre-Reviewed Chapter. Available
online: https://www.intechopen.com/chapters/73215 (accessed on 21 November 2021). [CrossRef]
50. Shobana, S.; Kumar, G.; Bakonyi, P.; Saratale, G.D.; Al-Muhtaseb, A.H.; Nemestóthy, N.; Bélafi-Bakó, K.; Xia, A.; Chang, J. A
review on the biomass pretreatment and inhibitor removal methods as key-steps towards efficient macroalgae-based biohydrogen
production. Bioresour. Technol. 2017, 244, 1341–1348. [CrossRef]
51. Cellulosic Yeast for G2 Ethanol Production. Available online: https://www.lallemandbds.com/wp-content/uploads/2015/06/
Xyloferm-product-sheet-2016-05-19.pdf/ (accessed on 22 September 2021).
52. Ciliberti, C.; Biundo, A.; Albergo, R.; Agrimi, G.; Braccio, G.; de Bari, I.; Pisano, I. Syngas Derived from Lignocellulosic Biomass
Gasification as an Alternative Resource for Innovative Bioprocesses. Processes 2020, 8, 1567. [CrossRef]
53. Renewable Fuels Association Analysis of Public and Private Data. Available online: https://ethanolrfa.org/markets-and-
statistics/annual-ethanol-production (accessed on 1 December 2021).
54. OECD-FAO Agricultural Outlook 2021–2030. Available online: https://www.fao.org/publications/oecd-fao-agricultural-
outlook/2021-2030/en/ (accessed on 12 October 2021).
55. Brazil Biofuels Annual Report 2020, Global Agricultural Information Network; GAIN Report Number BR2020-0032; Foreign Agricultural
Service, U.S. Department of Agriculture: Washington, DC, USA, 2020.
56. European Union Biofuels Annual Report 2021, Global Agricultural Information Network; GAIN Report Number E42021-0053; Foreign
Agricultural Service, U.S. Department of Agriculture: Washington, DC, USA, 2021.
57. China Biofuels Annual Report 2020, Global Agricultural Information Network; GAIN Report Number CH2020-0105; Foreign Agricul-
tural Service, U.S. Department of Agriculture: Washington, DC, USA, 2020.
Fermentation 2021, 7, 314 18 of 18

58. U.S. Bioenergy Statistics. Available online: https://www.ers.usda.gov/data-products/u-s-bioenergy-statistics/ (accessed on 22

November 2021).
59. Renewable Fuels Association. 2020 Ethanol Industry Outlook. Available online: https://ethanolrfa.org/file/21/2020-Outlook-
Final-for-Website.pdf/ (accessed on 12 October 2021).
60. New Energy Blue to Construct Cellulosic Biorefinery in Iowa. Available online: http://biomassmagazine.com/articles/18180/
new-energy-blue-to-construct-cellulosic-biorefinery-in-iowa/ (accessed on 12 October 2021).
61. Inbicon Biomass Refinery. Available online: https://www.eia.gov/biofuels/workshop/pdf/paul_kamp.pdf/ (accessed on 12
October 2021).
62. Argentina Biofuels Annual Report 2020, Global Agricultural Information Network; GAIN Report Number AR2020-0027; Foreign
Agricultural Service, U.S. Department of Agriculture: Washington, DC, USA, 2020.
63. Colombia Biofuels Annual Report 2021, Global Agricultural Information Network; GAIN Report Number CO2021-0012; Foreign
Agricultural Service, U.S. Department of Agriculture: Washington, DC, USA, 2021.
64. Nong, D.; Wang, C.; QuasemAl-Aminde, A. A critical review of energy resources, policies and scientific studies towards a cleaner
and more sustainable economy in Vietnam. Renew. Sustain. Energy Rev. 2020, 134, 110117. [CrossRef]
65. USDA Foreign Agricultural Service. Vietnam Ethanol Background Report. 11 August 2020. Available online: https://www.fas.
usda.gov/data/vietnam-vietnam-ethanol-background-report (accessed on 18 May 2021).
66. Kima, J.-S. Production of bioethanol from lignocellulose: Status and perspectives in Korea. Bioresour. Technol. 2010, 101, 4801–4805.
[CrossRef] [PubMed]
67. Reuter’s Report, China Set to Triple Its Ethanol Production Capacity: Government Researcher. Available online: https:
//www.reuters.com/article/us-china-ethanol/china-set-to-triple-its-ethanol-production-capacity-government-researcher-
idUSKBN1OA0FH (accessed on 21 May 2021).
68. India Biofuels Annual Report 2020, Global Agricultural Information Network; GAIN Report Number IN2021-0072; Foreign Agricultural
Service, U.S. Department of Agriculture: Washington, DC, USA, 2020.
69. Thailand Biofuels Annual Report 2020, Global Agricultural Information Network; GAIN Report Number TH2021-0040; Foreign
Agricultural Service, U.S. Department of Agriculture: Washington, DC, USA, 2020.
70. Australia Biofuels Annual Report 2020, Global Agricultural Information Network; GAIN Report Number AS2020-0020; Foreign
Agricultural Service, U.S. Department of Agriculture: Washington, DC, USA, 2020.
71. Bioethanol in Africa: The Case for Technology Transfer and South-South Cooperation. 2016. Available online: https://www.irena.
org/-/media/Files/IRENA/Agency/Publication/2016/IRENA_Bioethanol_in_Africa_2016.pdf (accessed on 1 December 2021).
72. Thomas, V.; Kwong, A. Ethanol as a lead replacement: Phasing out leaded gasoline in Africa. Energy Policy 2001, 29, 1133–1143.
[CrossRef]