PRETRATAMIENTOS (REDUCCIÓN DE LA LIGNOCELULOSA) PARA LA PRODUCCIÓN DE

BIOETANOL

1. Evaluation of ethanol production from sugar and lignocellulosic part of energy cane
(temperatura)

Preparation of energy cane bagasse hemicellulosic hydrolysate using dilute-acid

hydrolysis
The milled bagasse was dried at 60 °C for 24 h. The dried bagasse in a solid-liquid ratio
of 1:10 was soaked in 1% H2SO4 for 30 min at ambient temperature and then
autoclaved at 121 °C for 30 min. The hydrolysate was filtered and neutralized with CaO
to pH 5.5. The precipitate that formed was removed using centrifugation and filtration
(Senatham et al., 2016). Detoxification of energy cane bagasse hemicellulosic
hydrolysate was performed with slightly modified method described by Branco et al.
(2011). The neutralized hemicellulosic hydrolysate was treated with 2% (w/v) activated
charcoal with stirring at 150 rpm for 1 h at 30 °C. The solids were then removed using
vacuum filtration.
https://doi.org/10.1016/j.indcrop.2017.07.023

2. Industrial technologies for bioethanol production from lignocellulosic biomass

(explosión stean)

Pretreatment plays a significant role in bioethanol production from lignocellulosic

biomass [14,23,24]. Its objective is to disrupt the recalcitrance and heterogeneous
structure, increase the accessibility of hydrolytic enzymes, and reduce the inhibitors to
subsequent process [4,13]. To achieve the objective, it is essential to overcome the
intrinsic characteristic of lignocellulosic biomass, including recalcitrance, heterogeneity,
multi-composition, and diversity [59]. The intrinsic structural factors have been thought
as the direct resistance to the conversion of lignocellulosic biomass into ethanol [60].
Analysis of morphological structure showed that lignocellulosic biomass was essentially
a porous medium. Zhao and Chen reported that intrinsic structure of corn stover was a
porous medium with cell wall and middle lamella as skeleton, forming pore with
different diameters for the storage and transport of flow. Pores of corn stover can be
classified into five classes according to their morpha and diameter: molecule capillary
(3–11 nm), cell wall capillary (11–100 nm), cell lamellar capillary (100–1300 nm), cell
lumen capillary (1.3–50 μm) and particle capillary (50– 370 μm) [19]. The efficient
enzymatic hydrolysis means that the cellulase and hydrolysate can transfer effectively
within the porous structure. Thus the porous structure, especially pore size distribution
is largely responsible for the efficiency of enzymatic hydrolysis [61]. In order to increase
the economic viability, the pretreatment should aim to increase the effective porous
structure for enzymatic hydrolysis.
https://doi.org/10.1016/j.rser.2015.12.069

3. Techno-Economic Analysis of Different Pretreatment Processes for Lignocellulosic-

based Bioethanol Production
 2.1.1 Process setup for diluted acid pretreatment – DA
Figure 1 presents the setup of the diluted acid pretreatment ethanol process. In the
diluted acid pretreatment method, water was added to the raw material to dilute the
solids fraction to 30% (wt.) and the stream was heated in a presteamer to a temperature
of 100°C. Sulfuric acid at a ratio of 18 mg/g dry biomass was then mixed with the biomass
and the stream was sent to the pretreatment reactor. The reactor operates at 158°C
and 5.5 atm and the process has 5 minutes of residence time. The pretreated biomass
was then flashed at 130°C to remove part of the hydrolysis and fermentation inhibitors.
To the bottom stream of the flash tank 4.1 mg of sulfuric acid/g dry biomass were added
and the acidified stream was led to the oligomer conversion reactor where part of the
xylose oligomer was converted into xylose. The oligomer conversion reactor operates at
130°C and 5.7 atm and has a residence time of 30 minutes. The effluent of the reactor
was sent to a second flash tank, operating at atmospheric pressure, and then directed
to the conditioning tank. In the conditioning tank ammonia was added to correct the pH
of the mixture and more water was added to dilute the solids to 20% (wt.) in a
preparation for the hydrolysis step.
2.1.2 Process setup for liquid hot water pretreatment – LHW
Figure 2 presents the setup for the bioethanol production process based on the liquid
hot water pretreatment method. The biomass was mixed with water to keep the solids
loading content at 20% (wt.) and heated to 200°C by indirect steam heating. After 10
minutes of reaction holding time, the slurry was cooled down and sent to a flash tank to
remove part of the inhibitors. The bottom stream of the flash tank had its solid and liquid
phase separated by a pressure belt filter and a post-wash step was used to remove
impurities from the solid fraction. The solid and liquid phases were then mixed together
again and water was added to keep the solids level at 20% (wt.) prior to the hydrolysis.
6 Pretreatment and hydrolysis reactions and conversions were based on the work of Tao
et al. (2011).
2.1.3 Process setup for ammonia fiber explosion pretreatment – AFEX
Figure 3 presents the setup for the bioethanol production process based on the
ammonia fiber explosion pretreatment method. Initially water was added to the process
at a ratio of 0.81 to dry weight biomass and the slurry was sent to the pretreatment
reactor where ammonia was mixed at a ratio of 1.52 to dry weight biomass. The reactor
is operated at 150°C with 30 minutes of residence time. The outlet stream of the reactor
was quickly depressurized at the knock out drum and the liquid flashed, breaking the
structure of the lignocellulosic biomass. The flash process occurs at 105°C and the
overhead of the tank, containing concentrated ammonia was compressed, condensed
and sent to recycle in the process. After the knock out drum there is a second flash
operated at 85°C. The overhead of the flash tank was sent to an absorber column to
purify the ammonia before recycling to the process. The slurry from the flash tank had
its solid and liquid phase separated by a pressure belt filter and the solid phase went
through a washing step to further remove contaminants. The two phases were then
mixed and water was added to reach 20% (wt.) of solids fraction before the hydrolysis
and fermentation steps. The process setup, reactions and conversions were based on
Tao et al. (2011).
https://doi.org/10.1016/j.biortech.2016.07.007
4. Lignocellulosic bioethanol: A review and design conceptualization study of production
from cassava peels
 Pre-treatment (explosión de vapor)

Next to the biomass storage option, if not properly managed, this process step has the
propensity to increase the setup costs for a bioethanol processing plant. For the cassava
peel process, a catalyzed steam explosion is preferred over the LHW pretreatment
which requires more water and energy requirement and is therefore unfavorable for
commercial scale development. In making this selection, energy availability and water
requirements in the cultivating areas of cassava, with the potential of
commercialization, are the predominant factors. In the proposed process design, the
ground cassava peel can unloaded and subsequent delivery into a pressure chamber for
a catalyzed steam explosion (CSE) treatment. The CSE procedure is proposed for the
cassava peel pretreatment as it has been shown in literature that the addition of the
catalyst during this step can lead to a significant increase in sugar yields [104]. In this
procedure, steam at high temperature is introduced into the reactor for about 5 min
(usually at a temperature of about 433 K at a pressure of 1.5 MPa). The subsequent rapid
reduction in pressure causes an expansion of the lignocellulosic matrix, thereby leading
to a separation of individual fibers [13]. A dilute acid sulfuric acid catalyst can employed
as it is inexpensive and can also reduce the xylose content by as much as 90% [6,105],
all of which have positive effects on ethanol yield. The end of the exposure marks the
end of the cassava peels pretreatment stage and this is a very important stage ensure
an optimal yield of ethanol yield [106].
https://doi.org/10.1016/j.rser.2016.06.064

5. Effect of steam-pretreatment combined with hydrogen peroxide on lignocellulosic

agricultural wastes for bioethanol production: analysis of derived sugars and other by-
products
Steam pretreatment (explosión vapor)
Two different concentrations of H2O2-gas, corresponding to 1% and 0.2% by weight
based on the water content of SCB, were sprayed into raw material using a rotary
system, which allows a better surface area contact between the substrate and peroxide
solution [31]. Thus, the plastic bags containing impregnated materials were sealed and
kept for 2 h at room temperature. The steam explosion was performed in 10 L reactor
and the pretreated material was collected in a flash tank. The pretreatment unit is
shown in Fig. 1. Steam was provided using a 110 KW electrical boiler (Pann-Partner,
Stockholm, Sweden) able to provide steam up to a maximum pressure of 3 MPa,
equivalent to a saturated steam temperature of 235 °C [38]. The reactor included a ball
valve at the top for the feedstock input and an air-actuated ball valve positioned at the
bottom outlet, allowing for the swift expulsion of the pretreated material. Two different
air-actuated valves injected high-pressure steam into reactor, one of which (located
near to the bottom) was necessary for rapid heat up of the material, and the other
(located higher up) was required for temperature and pressure control. The
temperature, pressure and hold-up time in the reactor were monitored and regulated
by a computer, using the software Intouch (Wonderware, USA) [38]. The bagasse
equivalent to 600 grams of dry matter was loaded into reactor of steam explosion
system and treated for 15 min with a steam pressure of 1.89 MPa, corresponding to
temperatures of 210 °C, in according to literature data [31−33,39]. Before each
experiment, the reactor was first pre-heated in order to achieve the required
experimental temperature; time zero (for pretreatment) was taken when the pressure
 in the reactor reached 99 % of the target pressure. The pretreated-SCB was collected in
a flash tank and then stored at 5 °C in containers covered with plastic lids.

5.1. Determination of sugars and lignocellulose by-products concentrations

Sugars and lignocellulose by-products analysis was performed by HPLC (high performance
liquid chromatography), as described by Sluiter et al. [41]. The instrument used (Shimadzu LC-
10AD, Tokyo, Japan) was equipped with a Refractive Index detector (Shimadzu). All samples
were diluted and filtered through a 0.20 µm filter prior to HPLC analysis, and acidic samples
were neutralized by the addition of CaCO3. The concentrations of sugars, as cellobiose,
glucose, xylose, galactose, mannose and arabinose, in the liquid collected after pretreatment
and in the samples from enzymatic hydrolysis were separated using an Aminex HPX-87P
column (Bio-Rad laboratories, Hercules, CA, USA), operating at 85 °C with deionized water as
the mobile phase, at a flow rate of 0.6 mL/min. The concentrations of lignocellulose derived
by-products, as furfural, HMF, and acetic, levulinic, formic acid, were analyzed with an Aminex
HPX-87H (Bio-Rad laboratories, Hercules, CA, USA), operating at 65 °C, with 5 mM H2SO4 as
mobile phase at a flow rate of 0.6 mLl/min. The obtained data were subjected to analysis of
variance (ANOVA) for the model of sugars and lignocellulose by-products yields after hydrolysis
for pretreatment of SCB by steam and H2O2, in order to determine if any significant
differences (p < 0.05) occurred between factor. It can be seen that the model presents a high
correlation coefficient and can be considered statistically significant with 90% of confidence
according to the F test.
https://doi.org/10.1016/j.jechem.2017.11.007

6. Production of bioethanol from lignocellulosic materials via the biochemical pathway: A

review
6.1. Physico-chemical pretreatment
6.1.1. Steam explosion (autohydrolysis).
Steam explosion is initiated at a temperature of 433–533 K with a corresponding pressure
0.69–4.83 MPa for several seconds to a several minutes before the material is exposed to
atmospheric pressure for cooling [85].
Uncatalyzed steam explosion refers to a pretreatment technique in which lignocellulosic
biomass is rapidly heated by highpressure steam without addition of any chemicals. The
biomass/ steam mixture is held for a period of time to promote hemicellulose hydrolysis, and
terminated by an explosive decompression [86]. Negro et al. [87] studied steam explosion and
liquid hot water methods for pretreatment of poplar (Populus nigra) biomass. The best results
were obtained in steam explosion pretreatment at 483 K and 4 min, taking into account
cellulose recovery above 95%, enzymatic hydrolysis yield of about 60%, and 41% xylose
recovery in the liquid fraction. Addition of H2SO4 (or SO2) or CO2 [typically 0.3–3% (w/w)] in
steam explosion can decrease time and temperature, effectively improve hydrolysis, decrease
the production of inhibitory compounds, and lead to complete removal of hemicellulose [68].
H2SO4 is a strong catalyst that highly improves the hemicellulose removal but also easily yields
inhibitory substances [88]. Ballesteros et al. [89] applied acid-catalyzed steam explosion
pretreatment of wheat straw for bioethanol production by varying the temperature (433–473
K), the residence time (5, 10 or 20 min) and the acid concentration [H2SO4 0.9% (w/w)].
According to results of this study, the best pretreatment conditions to obtain high conversion
yield to bioethanol (approx 80% of theoretical) of cellulose-rich residue after steam explosion
are 463 K and 10 min or 473 K and 5 min, in acid-impregnated straw. Using a H2SO4-catalyzed
steam explosion process for pretreatment of Salix chips, at 473 K for either 4 or 8 min using
0.5% sulfuric acid, resulted in glucose recovery about 92% and 86% xylose recovery after
enzymatic hydrolysis [90]. SO2 appears more appealing than H2SO4 in steam explosion since
the former requires milder and much less expensive reactor material, generates less gypsum,
yields more xylose, and produces more digestible substrate with high fermentability [76]. The
treatment can be carried out by 1– 4% SO2 (w/w substrate) at elevated temperatures, e.g.
433–503 K, for a period of e.g. 10 min [56]. The main drawback of SO2 is its high toxicity, which
may pose safety and health risks. However, SO2 is used in various industrial processes using
established techniques [91].
Two-step pretreatment has been suggested in several studies as a means of increasing the
sugar recovery [92,93]. In the first step, steam is performed using low temperature to
solubilize hemicellu- losic fraction, and cellulose fraction is subjected to a second steam
explosion pretreatment step at a temperature higher than 483 K. It offers some additional
advantages (higher bioethanol yields, better use of raw material and lower enzyme dosages
during steam explosion) [81].

6.1.2. Ammonia fiber explosión

Ammonia fiber explosion (AFEX) is one of the alkaline physico-chemical

pretreatment processes. In this process, the material is subjected to liquid
ammonia at high temperature and pressure, and a subsequent fast
decompression, similar to the steam explosion, which causes a fast
saccharification of the lignocellulosic material [94]. In a typical AFEX process, the
dosage of liquid ammonia is 1–2 kg ammonia/kg dry biomass, the temperature is
363 K, and the residence time is 30 min [68,74]. The effective parameters in the
AFEX process are ammonia loading, temperature, water loading, blowdown
pressure, time, and number of treatments [56]. This system does not directly
liberate any sugars, but allows the polymers (hemicellulose and cellulose) to be
 attacked enzymatically and reduced to sugars [95]. AFEX pretreatment yields
optimal hydrolysis rates for pretreated lignocellulosics with close to theoretical
yields at low enzyme loadings (<5 FPU/g of biomass or 20 FPU/g cellulose)

AFEX pretreatment has been demonstrated to markedly improve the

saccharification rates of numerous herbaceous crops and grasses [78]. It has been
applied to various lignocellulosic materials, including rice straw, municipal solid
waste, newspaper, sugar beet pulp, sugarcane bagasse, corn stover, switchgrass,
miscanthus, apsen chips, etc. [76]. The optimal conditions for pretreatment of
switchgrass with AFEX were reported by Alizadeh et al. [96]. The optimal
pretreatment conditions were found to be near 373 K reactor temperature, and
ammonia loading of 1 kg of ammonia per kilogram of dry matter with 80%
moisture content (dry weight basis) at 5 min residence time. These conditions
yielded a sixfold improvement in hydrolysis efficiency. Teymouri et al. [97]
evaluated the optimum process conditions for the pretreatment of corn stover.
The optimal pretreatment conditions were found to be a temperature of 373 K, an
ammonia loading of 1 kg of ammonia per kilogram of dry matter, a moisture
content of 60% (dry weight basis), and a residence time of 5 min. Approximately
98% of the theoretical glucose yield was obtained during enzymatic hydrolysis of
the optimal treated corn stover using 60 filter paper units (FPU) of cellulase
enzyme/g of glucan (equal to 22 FPU/g of dry corn stover). The bioethanol yield
from this sample was increased up to 2.2 times over that of untreated sample. The
composition of the materials after AFEX pretreatment was essentially the same as
the original materials [74]. Holtzapple et al. [98] obtained over 90% of hydrolysis of
cellulose and hemicellulose after AFEX pretreatment of Bermuda grass
(approximately 5% lignin) and bagases (15% lignin). AFEX works only moderately
and is not attractive for the biomass with high lignin content. Lignin content in
grasses (15–20%) is relatively low when compared with hardwoods and softwood
(20–35%) [98]. This could be one of the major reasons that grasses can be more
easily digested compared to AFEX treated hardwoods following AFEX pretreatment
[99]. Ammonia must be recycled after the pretreatment to reduce the cost and
protect the environment [56]. A possible approach is to recover the ammonia after
the pretreatment by evaporation

6.1.3. Liquid hot-water pretreatment.

Cooking of lignocellulosic materials in liquid hot water (LHW) is one of the

hydrothermal pretreatment methods applied for pretreatment of lignocellulosic
materials since several decades ago in e.g. pulp industries [56]. LHW subjects
biomass to hot water in liquid state at high pressure during a fixed period and it
presents elevated recovery rates for pentoses and generates low amount of
inhibitors [81]. This pretreatment process usually has involved temperatures of
473- 503 K for up to 15 min. Around 40–60% of the total mass is dissolved in this
process, with 4–22% of the cellulose, 35–60% of the lignin and all of the
hemicellulose being removed [101]. If the pH is maintained between 4 and 7, the
degradation of monosaccharide sugars can be minimized [102].

6.2. Chemical pretreatment

6.2.1. Ozonolysis.
Ozonolysis involves using ozone gas to break down the lignin and hemicellulose
and increase the biodegradability of the cellulose. The pretreatment is usually
carried out at room temperature and is effective at lignin removal without the
formation of toxic by-products [103]. Ozonation has been widely used to reduce
the lignin content of both agricultural and forestry wastes [104]. Ozonolysis has
been shown to break down 49% of lignin in corn stalks and 55–59% of lignin in
autohydrolyzed (hemicellulose free) corn stalks [105]. In a study [106], wheat and
rye straws were pretreated with ozone to increase the enzymatic hydrolysis extent
of potentially fermentable sugars. Enzymatic hydrolysis yields of up to 88.6% and
57% were obtained compared to 29% and 16% in non-ozonated wheat and rye
straw respectively. A drawback of ozonolysis is that a large amount of ozone is
required, which can make the process expensive [68].

6.2.2. Alkaline pretreatment.

Alkali pretreatment refers to the application of alkaline solutions to remove lignin

and various uronic acid substitutions on hemicellulose that lower the accessibility
of enzyme to the hemicellulose and cellulose [107,108]. These processes utilize
lower temperatures and pressures compared to other pretreatment technologies.
Alkali pretreatment may be carried out at ambient conditions, but pretreatment
time is measured in terms of hours or days rather than minutes or seconds [86].
Regardless the advantages, these methods present difficulties from the point of
view of the process economy for obtaining fuels [94]. Sodium, potassium, calcium
and ammonium hydroxide are appropriate chemicals for pretreatment. Of these
four, NaOH has been studied the most [68]. Dilute NaOH treatment of
lignocellulosic biomass causes swelling, leading to an increase in the internal
surface area, a decrease in crystallinity, separation of structural linkages between
lignin and carbohydrates, and disruption of the lignin structure [109]. Millet et al.
[110] reported that the digestibility of NaOH-treated hardwood to increase from
14% to 55% with the decrease of lignin content from 24%–55% to 20%. However,
no effect of dilute NaOH pretreatment was found for softwoods with lignin
content greater than 26%. Silverstein et al. [107] reported >65% lignin reduction in
cotton stalk treated with 2% NaOH for 90 min at 394 K/15 psi. Cellulases produced
by Bacillus subtilis for the saccharification of wheat straw, rice straw and bagasse
were used by Akhtar et al. [111]. Pretreatment of these substrates with 2% NaOH
was found to be more effective for increasing the saccharifi- cation. The
saccharification rates of 33.0%, 25.5% and 35.5% were obtained with 2% NaOH
pretreated wheat straw, rice straw and bagasse, respectively. In a study [112], a
combination of NaOH treatment and homogenization was used as a pretreatment
to enhance the enzymatic hydrolysis of corn stover. The highest glucose yield (6.25
g/l) was obtained when the corn stover was pretreated by a combination of 1.0 N
NaOH treatment and homogenization (Fig. 5).

Lime (Ca(OH)2) as compared to NaOH and KOH has lower cost and less significant
safety requirements. It can be recovered from hydrolyzate by reaction with CO2,
so that formed carbonate can then be reconverted to lime [113]. To make lime as
efficient as other alkalis in enhancing the digestibility of lignocellulose, appropriate
pretreatment conditions need to be employed [114]. Lime, water, and an oxidizing
agent (air or O2) are mixed with the biomass at temperatures ranging from 313 to
423 K for a period ranging from hours to weeks [115]. Two types of lime treatment
have been explored: (1) short-term and (2) long-term. Short-term lime
pretreatment involves boiling the biomass with a lime loading of 0.1 g Ca(OH)2/g
dry biomass at temperatures of 358–408 for 1– 3 h [116]. Long-term pretreatment
involves using the same lime loading at lower temperatures (313–328 K) for 4–6
weeks in the presence of air [116]. Lime has been used to pretreat switchgrass
(373 K for 2 h) [117], wheat straw (394 K for 1 h) [118], corn stover (373 K for 13 h)
[119], and poplar wood (423 K for 6 h with 14-atm oxygen) [120]. Saha and Cotta
[118] obtained maximum total sugar yield (451 ± 3mg g1 straw; glucose, 252 ± 6
mg; xylose, 173 ± 3 mg; arabinose, 27 ± 2 mg; 65% conversion) by lime
pretreatment (100mg g1 straw, 394 K, 1 h). The authors also investigated the
effects of pH (3.5–6.5) and temperature (298–343 K) on the enzymatic hydrolysis
of lime pretreated wheat straw (8.6%, w/v) using a combination of three enzymes
(cellulose, b-glucosidase, and hemicellulase), each enzyme at a dose level of 0.05
ml g1 substrate. Fig. 6 shows the effect of pH and temperature on the enzymatic
hydrolysis of lime pretreated wheat straw. Alkaline peroxide is one of the effective
pretreatment methods that can improve the enzymatic hydrolysis by
delignification of lignocellulosicmaterials. In this method, the lignocelluloses are
soaked in pH-adjusted water (e.g. to pH 11–12 using NaOH) containing H2O2 at
room temperatures for a period of time (e.g. 6–24 h) [56].

6.2.3. Acid pretreatment.

Acid pretreatments normally aim for high yields of sugars from lignocellulosic
materials. Acid pretreatment involves the use of sulfuric, nitric, or hydrochloric
acids to remove hemicellulose components and expose cellulose for enzymatic
digestion [107]. The acid pretreatment can operate either under a high
temperature and low acid concentration (dilute acid pretreatment) or under a low
temperature and high acid concentration (concentrated acid pretreatment) [56].
Dilute acid hydrolysis has been successfully developed for pretreatment of
lignocellulosic materials. The dilute acid pretreatment works fairly well on
agricultural feedstocks, such as corn stover and rice/wheat straw [62]. While dilute
acid pretreatments are known to improve enzymatic hydrolysis, their cost is
relatively high compared to physico-chemical pretreatments [44]. This
pretreatment method gives high reaction rates and significantly improves cellulose
hydrolysis [47]. There are primarily two types of dilute acid pretreatment
processes: low solids loading (5–10% [w/w]), high temperature (T > 433 K),
continuous-flow processes and high solids loading (10–40% [w/w]), lower
temperature (T < 433 K), batch processes [121]. In general, higher pretreatment
temperatures and shorter reactor residence times result in higher soluble xylose
recovery yields and enzymatic cellulose digestibility. Higher-temperature dilute
acid pretreatment has been shown to increase cellulose digestibility of pretreated
residues [122]. Depending on the substrate and the conditions used, between 80%
and 95% of the hemicellulosic sugars can be recovered by dilute acid pretreatment
from the lignocellulosic material [47,123,124]. Silverstein et al. [107] reported 95%
xylan reduction in cotton stalk treated with 2% H2SO4 for 90 min at 394 K/15 psi

In recent years, treatment of lignocellulosic biomass with dilute sulfuric acid has
been primarily used as a means of hemicellulose hydrolysis and pretreatment for
 enzymatic hydrolysis of cellulose [125]. Sulfuric acid at concentrations usually
below 4 wt.%, has been of the most interest in such studies as it is inexpensive and
effective [68]. Dilute sulfuric acid pretreatment (0.2–2.0% sulfuric acid, 394–493 K)
of lignocellulose serves three important functions in the conversion process [121]:
(1) hydrolysis of the hemicellulose components to produce a syrup of monomeric
sugars, (2) exposure of cellulose for enzymatic digestion by removal of
hemicellulose and part of the lignin, and (3) solubilization of heavy metals which
may be contaminating the feedstock. In spite of these benefits, dilute sulfuric acid
has some important disadvantages [76]: (1) corrosion that mandates expensive
materials of construction, (2) acidic prehydrolyzates must be neutralized before
the sugars proceed to fermentation, (3) gypsum has problematic reverse solubility
characteristics when neutralized with inexpensive calcium hydroxide, (4) formation
of degradation products and release of natural biomass fermentation inhibitors
are other characteristics of acid pretreatment, (5) disposal of neutralization salts is
needed, and (6) biomass particle size reduction is necessary.

6.3. Biological pretreatment

Biological pretreatment involves microorganisms such as brown-, white- and soft-

rot fungi that are used to degrade lignin and solubilize hemicellulose. White-rot
fungi are the most effective biological pretreatment of lignocellulosic materials
[56,68,74]. Lee et al. [126] evaluated biological pretreatment of Japanese red pine
(Pinus densiflora) using three white-rot fungi (Ceriporia lacerata, Stereum
hirsutum, and Polyporus brumalis). pretreatment with S. hirsutum resulted in
selective degradation of the lignin rather than the holocellulose component. The
advantages of biological pretreatment include low energy requirement and mild
environmental conditions. However, the rate of hydrolysis in most biological
pretreatment process is very low [74].

https://doi.org/10.1016/j.enconman.2010.08.013