Name: Date:

Course Code and Section: Course Degree and Year:

CONCEPTUAL LITERATURE SECTION EXERCISE 3

Directions: Identify the techniques used in organizing ideas of the following Conceptual Literature section
by surrounding each sub-section with brackets and extending each of the brackets with arrows leading to the
writing of their specific techniques on the margins.

2.1 Corn Cobs

Corns, botanically known as Zea mays, are cereal plant grains that serve as major crops commonly cultivated
among tropical countries especially in the Philippines (Arias, 2017). Corn cob is the part of the corn where the kernels
grow and serves as the core which holds the grains of a corn (“corncob,” 2016). It is an elongated and cylindrical part
of an ear of corn, which also has woody properties that makes it hard and thick (“corncob,” 2016).
In terms of chemical composition, corn cobs contain essential constituents for biochemical processing. Corn
cobs, as lignocellulosic agricultural wastes, contain 45% cellulose, 35% hemicellulose and 15% lignin (Fortunati et al.,
2016). Figure 2.1 illustrates the common wood structure with these three constituents.

Figure 2.1 Usual Woody Material Structure of (A) Cellulose, Lignin and
Hemicellulose, and (B) Cellulose alone (Fortunati et al., 2016)

Fortunati et al. (2016) state that cellulose is a polymer that is considered as one of the most abundant and
naturally-occurring among its kind and it has been known as a plausible raw material biochemical processing.
Fortunati et al. add that hemicellulose, on the other hand, is a polysaccharide found in plant cell walls that includes
xylans, mannans and glucomannans. Meanwhile, lignin is an aromatic polymer that forms a layer covering around the
two other parts that provides sturdiness to the plant; however, lignin is generally considered to be a waste material in
most processing (Fortunati et al., 2016). Lignin is identified as a major limiting factor among all biomass components
(Barathikannan & Agastian, 2016).
Lignocellulosic materials such as corn cobs are known for being renewable and natural and having promising
potentials in the modern industrial and biochemical societies, as large amounts of similar biomass may be used in
production of valuable products (Fortunati et al., 2016).

2.2 Acid Hydrolysates

Saccharification is a process of breaking down the hemicellulose, such as xylan, into monosaccharide
components, such as xylose (Arias, 2017). Acid hydrolysis is one method of saccharification that uses an acid in the
conversion of biomass producing high amounts of xylose in the hydrolysate (Arias, 2017). Dilute sulfuric acid is
usually used in acid hydrolysis.
In the production of high-value products, lignocellulosic materials must first be hydrolyzed (Jeevan et al.,
2011). Lignocellulosic materials with high xylan content undergo hydrolysis in order to produce the hydrolysates.
Specifically, acid hydrolysates contain hexoses such as glucose, acetic acid, hydroxymethylfurfural (HMF), furfurals,
and other phenolic compounds, aside from the desired pentose sugars such as xylose and arabinose (Vallejos &
Area, 2017). The acid hydrolysates, due to its mixed sugars content, serve as substrate for fermentation of sugar-
utilizing organisms to produce value-added products such as ethanol and xylitol (Latif & Rajoka, 2001).

2.3 Xylitol and Its Production

According to Barathikannan and Agastian (2016), xylitol is a sugar alcohol of the five-carbon type, which
means one mole contains five carbon atoms and five hydroxyl groups. It has many medical and health benefits. Xylitol

1
Name: Date:
Course Code and Section: Course Degree and Year:

CONCEPTUAL LITERATURE SECTION EXERCISE 3

Directions: Identify the techniques used in organizing ideas of the following Conceptual Literature section
by surrounding each sub-section with brackets and extending each of the brackets with arrows leading to the
writing of their specific techniques on the margins.

serves as a sweetener for diabetic patients due to its non-cariogenic properties and non-fermentability and its
sweetness level is comparable with sucrose (Barathikannan & Agastian, 2016). Moreover, consumption of xylitol is
associated with several beneficial health effects such as significant reduction in tooth decay, increased bone density,
weight loss and stabilization of blood sugar level (Barathikannan & Agastian, 2016).
Xylitol can be produced by natural extraction, chemical processing, or biochemical production. Xylitol can be
obtained from fruits and vegetables naturally containing xylitol, such as raspberry, strawberry and yellow plum
(Barathikannan & Agastian, 2016). However, the extraction process is not cost-effective due to low xylitol contents of
these natural sources (Barathikannan & Agastian, 2016). On the other hand, Figure 2.2 shows the general procedures
in chemical and biochemical production processes as suggested by Vallejos and Area (2017).

Figure 2.2 General Xylitol Production Process (Vallejos & Area, 2017)

As seen in the figure above, xylitol production through chemical processing involves catalytic
dehydrogenation of pure xylose (Barathikannan & Agastian, 2016; Vallejos & Area, 2017). It is costly and requires a
lot of energy, and imposes environmental risk due to the toxic catalyst and the presence of high-pressure hydrogen
gas (Barathikannan & Agastian, 2016; Vallejos & Area, 2017). Afterwards, it undergoes downstream processing to
obtain xylitol of high purity.
As shown also in Figure 2.2 is the biotechnological pathway or the microbial production of xylitol from
agricultural wastes. After detoxifying the hydrolysates containing the target sugar (xylose), a microorganism is added
to proceed with fermentation. The use of yeast strains such as the Candida species has recently attracted much
attention (Barathikannan & Agastian, 2016; Horiuchi & Tada, 2005). According to Barathikannan and Agastian (2016)
and Vallejos and Area (2017), xylitol production from microorganisms has been considered sustainable because the
process can be conducted under mild controlled conditions and can produce xylitol a lot cheaper than other methods
with low environmental impact and high sustainability.

2.4 Detoxification
Detoxification is a method where inhibitors of fermentation by microorganisms are removed. Inhibitors are
compounds that can limit the uptake of xylose, reduce the growth of microorganisms, or totally stop the fermentation
from occurring.
The main inhibitors in the microbial production of xylitol with acid hydrolysates as substrate are: (1) mineral
deposits from biomass itself or from the corrosion of the equipment; (2) hemicellulose degradation products such as
furfuralthe phenolic compounds and aldehyde aromatic compounds; and (4) derivatives of extractives (Vallejos &

2
Name: Date:
Course Code and Section: Course Degree and Year:

CONCEPTUAL LITERATURE SECTION EXERCISE 3

Directions: Identify the techniques used in organizing ideas of the following Conceptual Literature section
by surrounding each sub-section with brackets and extending each of the brackets with arrows leading to the
writing of their specific techniques on the margins.

Area, 2017). Detoxification and pretreatment methods, which can involve physical, chemical, biological, and combined
methods, provide better accessibility to the necessary compounds after the hydrolysis of hemicellulose by overcoming
the complex structure of lignocellulosic biomass to allow the fermentation of usable sugars (Vallejos & Area, 2017).
Detoxification can involve physical, chemical or biological methods, or a combination of any of these.
“Physical methods mainly include evaporation, stream stripping, solvent extraction, and membrane separation”
(Vallejos & Area, 2017, p. 416). Physical methods include vacuum evaporation and stream stripping. Both methods
remove volatile compounds (acetic and formic acid) by more than 50% and a 100% removal of furfural, as well
increasing the amount of xylose necessary for biotechnological purposes (Vallejos & Area, 2017).
Vallejos and Area (2017) describe the chemical methods as methods that generally involve selective removal
of inhibitors by extraction by organic solvents. These organic solvents could either be ethyl acetate and similar
compounds or alkaline chemicals. According to Vallejos and Area, ethyl acetate totally eliminates the presence of
furfural, phenolic compounds, while trichloroethylene and benzene are the best solvents for acetic acid removal.
Moreover, various alkaline chemicals (NaOH, Ca(OH)2, NH OH, Na SO3, and others) are used in neutralization
methods which change the pH of the hydrolysates from acidic to neutral (pH 5.5–6.0). Adsorption by activated
charcoal is also classified under chemical methods and is an effective and low-cost method that specifically removes
HMF, furfural, and phenolic compounds, without xylose loss, while adsorption by ion exchange is used to remove
ions, both organic and inorganic, because of its desalination capability (Vallejos & Area, 2017).
Vallejos and Area (2017) add biological methods to the list and describe it as methods that are “based on the
use of microorganisms for the selective removal of inhibitors (S. cerevisiae mutant, Trichoderma reesei, others). They
were generalized to eliminate a given type of inhibitor, but the adaptation of the microorganism can be limited” (p.
418).
Detoxification by combined methods may include any of the previously discussed methods done after
another. An example is vacuum evaporation together with activated charcoal which further decreases the
concentration of acetic acid, HMF, furfural, and phenolic compounds (Vallejos & Area, 2017).

2.5 Flask Fermentation

Fermentation is a process of allowing a microorganism to grow in a medium (substrate) having its usual
metabolism that results in either cell growth, formation of biological products, or both. There are many ways to run a
fermentation process, one of which is the use of shake flask fermentation (Hassan, 2015).
Hassan (2015) states that shake flask fermentation is an example of batch fermentation where
microorganisms are grown and cultured in a flask, usually in an Erlenmeyer flask. Compared to a large-scale
fermenter, shake flask is similar to a stirred-tank bioreactor, having only small volume requirement of inoculum and
growth medium as major differences (Hassan, 2015).
Operating conditions are also important to be monitored (Klinger, Baumann, Puskeiler, & Jockwer, n.d.). It is
important to monitor the temperature, shaking frequency and incubation time, as well as aeration in order to attain the
desired growth rate (Hassan, 2015; Klinger et al., n.d.). Klinger et al. added that disturbances may affect the
metabolism and overall performance of the cultivated cell. Moreover, the aim of shaking is to keep the medium and
the culture homogeneous (i.e., well-mixed) and also to ensure that there is a sufficient aeration for aerobic conditions
(Hassan, 2015).
Fermentation studies deal mostly with cell kinetics. Cell kinetics is how the cells grow with respect to time.
The medium or substrate is initially inoculated with the microorganism. It is expected that the growth keeps on
increasing until inhibited due to decreasing substrate or toxic compounds. In this study, the substrate would be the
corn cob hydrolysates and the microorganism comes from the Candida sp.