CHAPTER 2

Review of Related Literature and Studies

2.1 RELATED LITERATURE

2.1.1 Cassava

Cassava(Manihot esculenta), sometimes also called manioc, is the third largest

source of carbohydrates for human consumption in the world, with an estimated
annual world production of 208 million tonnes. In the Philippines, it is commonly
called as kamoteng kahoy or balinghoy. Cassava is extensively cultivated as an annual
crop in tropical and subtropical regions for its edible starchy tuberous root, a major
source of carbohydrates. Cassava's starchy roots produce more food energy per unit of
land than any other staple crop. It is a major source of low cost carbohydrates and a
staple food for 500 million people in the humid tropics. On infertile land where the
cultivation of other crops is difficult, unless considerable inputs are applied, cassava
still has a reasonable yield. (Kuiper et. al, 2007)

Cassava thrives better in poor soils than in any other major food plant. As a
result, fertilization is rarely necessary. However, yields can be increased by planting
cuttings on well drained soil with adequate organic matter. Cassava is a heat-loving
plant that requires a minimum temperature of 80oF to grow. Since many cultivars are
drought resistant, cassava can survive even during the dry season when the soil
moisture is low, but humidity is high.

The plant grows tall, some reaching 15 feet, with leaves varying in shape and
size. The edible parts are the tuberous root and leaves. The tuber (root) is somewhat
dark brown in color and grows up to 2 feet long. (Sugino and Mayrowani, 2009)

The crop is highly efficient in producing starch, is available all year round, is
tolerant to extreme stress conditions, and fits nicely within traditional farming systems.
Fresh roots contain about 30% starch. Cassava starch is one of the best fermentable
substances for the production of ethanol. At the moment, sugarcane is the most widely

8
used crop for bio-ethanol in the tropics, but sugarcane requires a lot of water (Ado et.
al, 2009). Therefore, cassava is the best option as a feedstock for bio-ethanol
production since it can be grown easily and with low maintenance. Aside from that, a
much larger area in the tropics is available and suitable for growing cassava.

In each locality in the Philippines where the crop is grown, numerous cassava
cultivars exist, with different leaf sizes, plant heights, colors, tuber shapes, timings of
maturity, overall yields, dry matter contents, starch contents and cyanogenic glycoside
contents of the roots. Roots with irregular shapes are more difficult to harvest and to
peel, resulting in greater losses of usable root material. (Apea-Bah et. al, 2009)
Traditionally, cassava roots are processed by various methods into numerous products,
which are utilized in various ways according to local preferences.

2.1.2 Starch

Starch or amylum is a carbohydrate consisting of a large number of glucose

units joined together by glycosidic bonds. It is a natural organic polymer manufactured
by green plants through photosynthesis to serve as metabolic reserve. It is the most
important carbohydrate in the human diet and is contained in such staple foods as
potatoes, wheat, maize (corn), rice, and cassava. In grains, starch is contained in a
husk of cellulose. Precisely speaking, starch is basically a complex carbohydrate called
polysaccharide and is composed of several thousand glucose elements. Starch also
contains amylose and amylopectin and is the most important resource for food for
humans (Dean, 2000). Depending on the plant, starch generally contains 20 to 25%
amylose and 75 to 80% amylopectin (Mcmurry, 2003). Glycogen, the glucose store of
animals, is a more branched version of amylopectin.

Amylose is a linear polymer made up of D-glucose units. This polysaccharide

is one of the two components of starch, making up approximately 20-30% of the
structure. Because of its tightly packed structure, amylose is more resistant to
digestion than other starch molecules and is therefore an important form of resistant
starch which has been found to be an effective prebiotic. (Dean, 2000)

9
 Figure 2.1: The molecular structure of amylose

Amylopectin is a soluble
polysaccharide and highly branched
polymer of glucose found in plants
(Mcmurry, 2003). Starch is made of about
70% amylopectin by weight, though it
varies depending on the source (higher in
medium-grain rice till 100% in waxy rice,
waxy potato starch and waxy corn and
lower in long-grain rice, amylomaize and
russet potato, for example). Amylopectin
is highly branched, being formed of 2,000 Figure 2.2: The molecular structure of amylopectin
to 200,000 glucose units. (Dean, 2000).

Starch, which is mainly composed of amylopectin and amylase, can be

converted to alcohol by means of fermentation. Since certain organisms like yeast
cannot directly ferment the starch into alcohol, the starch must be broken down first
into simple sugars in the process call hydrolysis. The acid or enzyme hydrolysis of
starch produces two simple sugars of glucose which is consumed by yeast to produce a
by-product which is ethanol and carbon dioxide. This conversion of starch to alcohol
can be utilized as a renewable source of bioethanol since a huge variety of crops
contains a high percentage of starch. The development of this kind of technology helps
sustain the increasing demand for energy in this world.

10
2.1.3 Alcohols

Alcohol is commonly thought of as either rubbing alcohol, the active

ingredient in an alcoholic drink, or the additive that makes gasoline into gasohol.
Chemists generalize its meaning to include almost any carbon-hydrogen compound
with at least one hydroxyl group (symbolized as -OH) in its molecular structure
(Brewster et. al., 1997). Categorized by the number and placement of the -OH groups,
and the size and shape of the attached carbon molecule, alcohols are fundamental to
organic chemical synthesis. There are different kinds of alcohols depending on how
the OH group is positioned on the chain of carbon atoms. There are some chemical
differences between the various types such as its solubility and boiling points.

Alcohols are produced industrially from petroleum, coal, or other natural

products. Both ethanol and methanol can be obtained from petroleum or natural gas,
ethanol may be the most interesting because it can be obtained from a renewable
resource, particularly from organic material such as grain or cellulosic materials (Azmi
et.al, 2010). The "cracking" of crude petroleum yields many lower-molecule-weight
chemical compounds, including some starting materials for alcohols such as ethylene
and propylene. Ethylene reacts with hot steam over a catalyst to yield ethanol directly.
A process known as hydration produces isopropyl alcohol when water is chemically
added to propylene (Shreve, 1984). Alcohol can also obtain synthetically, via ethene or
acetylene, from calcium carbide, coal, oil gas, and other sources. In addition, fuel
alcohols can be produced from a variety of crops, such as sugarcane, sugar beets,
maize, barley, potatoes, cassava, sunflower, eucalyptus through the process of
fermentation (Cortes-Camidero, 2010).

Fermentation, one of the oldest chemical process known to man, is used to

make a variety of products, including foods, flavorings, beverages, pharmaceuticals,
and chemicals. At present, however, many of the simpler products such as ethanol are
synthesized from petroleum feedstock at lower costs. Alcohol fermentation is done by
yeast and some kinds of bacteria. These microorganisms convert sugars in ethyl
alcohol and carbon dioxide. Alcoholic fermentation begins after glucose enters the

11
cell. The glucose is broken down into pyruvic acid. This pyruvic acid is then converted
to CO2, ethanol, and energy for the cell. Fermentation process from any material that
contains sugars can derive ethanol. The many and varied raw materials used in the
manufacture of ethanol via fermentation are conveniently classified under three types
of agricultural raw materials: sugars, starches, and cellulose materials. Sugars (from
sugar cane, sugar beets, molasses and fruits) can be converted to ethanol directly.
Starches (from grains, potatoes, root crops) must first be hydrolyzed to fermentable
sugars by the action of enzymes from malt or molds. Cellulose (from wood,
agricultural residues, waste sulfite liquor from pulp and paper mills) must likewise be
converted to sugars, generally by the action of mineral acids. Once simple sugars are
formed, enzymes from yeast can readily ferment them to ethanol (Lim et.al, 2002).

Alcohols have applications in industry and science as reagents or solvents. One

of the examples of alcohols is ethanol which is widely used as solvent. Ethanol,
because of its low toxicity and ability to dissolved non-polar substance, can be used as
a solvent in medical drugs, perfumes, and vegetable essences such as vanilla. In
addition to its presence in alcoholic beverages, ethanol also is used as a solvent for
food extracts such as vanilla, perfumes, cosmetics, and some types of paints and
lacquers. It is relatively safe, and can be used to dissolve many organic compounds
which are insoluble in water. Isopropyl alcohol is widely used in industry as a solvent
for paints and chemical processes. Most alcohols, including ethanol, burn in air and
are frequently used as fuel. Methanol can sometimes be used as a fuel, though it
evaporates too quickly for regular use in cars (Zhang and Feng, 2010).

Agricultural alcohol for fuel requires substantial amounts of cultivable land

with fertile soils and water. However, if the fuel alcohol is made of the stalks, wastes,
clippings, straw, corn cobs, and other crop field trash, then no additional land is
needed. Using these sources for this purpose, however, would require additional
replacement in animal feedstock, fertilizers and electric power plant fuels (Eggeman,
2010). Using alcohol as a renewable source of energy has a great impact not only in
our economy but also to our environment. It helps sustain energy for our combustion
engines and preserve the natural habitats in our environment.

12
2.1.4 Tests for Alcohols

Lucas Test

The Lucas test is a test in alcohols that is conducted to test and differentiate
between the types of primary, secondary or tertiary alcohol. It uses the differences in
reactivity of hydrogen halides and the three classes or types of alcohol. In the reaction
the hydroxyl group is replaced by chlorine. The time it takes in the appearance of
turbidity is used as a measure for determining the class of alcohol. (Grolier, 2010)

Hence, the time taken for turbidity to appear is a measure of the reactivity of
the class of alcohol with Lucas reagent, and this is used to differentiate between the
three classes of alcohols:
no visible reaction: primary, such as normal amyl alcohol
solution turns cloudy in 3-5 minutes: secondary, such as sec-amyl alcohol
solution turns cloudy immediately, and/or phases separate: tertiary, such as tert-
amyl alcohol (2-Methyl-2-butanol)
The test is usually conducted at room temperature.

Chromic Acid Test

Chromic acid is capable of oxidizing many kinds of organic compounds and

many variations on this reagent have been developed. Chromic acid in aqueous
sulfuric acid and acetone is known as the Jones reagent, which will oxidize primary
and secondary alcohols to carboxylic acids and ketones respectively, while rarely
affecting unsaturated bonds. Tertiary alcohol groups are unaffected. Because of the
oxidation is signaled by a color change from orange to a blue-green, chromic acid is
used as a qualitative analytical test for the presence of primary or secondary alcohols.
(Acid test, 2010)

13
Iodoform Test

Compounds with a methyl group next to a carbonyl group give a positive result
with the iodoform (tri-iodomethane) test. Ethanol and secondary alcohols with a
methyl group attached to the same carbon as the OH group will also give a positive
iodoform test. This is because the iodine oxidizes the alcohols to a carbonyl compound
with a methyl group next to the carbonyl group.

When iodine and sodium hydroxide are used as the reagents, a positive reaction
gives iodoform. Iodoform (CHI3) is a pale yellow substance. Due to its high molar
mass due to the three iodine atoms, it is solid at room. It is insoluble in water and has
an antiseptic smell. A visible precipitate of this compound will form from a sample
only when either a methyl ketone, ethanal, a methyl secondary alcohol or ethanol are
present. (Brewster et.al, 1977)

2.1.5 Fermentation

In a general sense, fermentation is the conversion of a carbohydrate such as

sugar into an acid or an alcohol. More specifically, fermentation can refer to the use of
yeast to change sugar into alcohol or the use of bacteria to create lactic acid in certain
foods. Fermentation occurs naturally in many different foods given the right
conditions, and humans have intentionally made use of it for many thousands of years.

Fermentation is a chemical process and produces heat. In concentrated or

particularly large mashes, the temperature can actually rise to levels dangerous to
yeast. Since the ideal temperature for yeast is around 85 oF, its best to maintain the
temperature by either utilizing cooling coils. (Yah et.al, 2010)

Fermentation process produces valuable products like ethanol which is used in

alcoholic beverages, as a solvent, and as a fuel. When certain species of yeast
(Saccharomyces cerevisiae) metabolize sugar, they produce ethanol and carbon
dioxide. The chemical equation below summarizes the conversion (Ocloo et. al, 2010):

C6H12O6 2 CH3CH2OH + 2 CO2

14
 Conversion of sugars to alcohol and CO2 will complete in three to five days,
depending on the temperature of the mixture and the type of yeast used. You can tell
when the mash is done by watching the cap of soils on top of the solution. During
fermentation, the rising of CO2 keeps the solids in constant motion, but when the
bubbling stops, the solids fall to the bottom. At this time, youre ready to separate the
solids from the liquids and distillation is employed to separate the alcohols from other
liquids and increase the purity of the ethanol.

2.1.6 Yeast

Yeast is a group of single celled fungi, a few species of which are commonly
used to leaven bread and ferment alcoholic beverages. More than one thousand species
of yeasts have been described. The most well known and commercially significant
yeast are the related species and strains of Saccharomyces Cervisiae. These organisms
have long been used to ferment the sugars of rice, wheat, barley, and corn to produce
alcoholic beverages and in baking to expand, or raise, dough. (Grolier, 2000)

In the absence of oxygen, fermentative yeasts produce their energy by

converting sugars into carbon dioxide and ethanol. This energy is necessary to sustain
life, from the oxidation of organic substances by molecular oxygen, in the process of
respiration. Under anaerobic conditions (low oxygen concentrations), many organisms,
including yeast, obtain the energy from the process of fermentation (Bailey, 1986). In
alcoholic fermentation, characteristic of many yeast species, the fermentation process
starts with one molecule of the six carbon sugar - glucose, and terminates with two
molecules of the two carbon alcohol - ethanol, and two molecules of CO2.

Yeast plants can propagate in a solution with or without air so agitated only
enough to saturate the wort with air and then let it stand still. If the mash is continually
agitated, the yeast will reproduce faster and make less waste: carbon dioxide and
alcohol. But if the solution becomes anaerobic (without air) the yeast slows down
reproduction and makes more alcohol and carbon dioxide. (Shuler et.al, 2002)

15
 Yeast also produces enzymes of its own to convert complex sugars. Since sugar
conversion and alcohol conversion takes place simultaneously, the amylase enzymes
and the yeast work in cooperation to convert dextrins to glucose and fructose and then
to alcohol and carbon dioxide.

2.1.7 Co-product Yields

Alcoholic fermentation begins after glucose enters the cell. The glucose is
broken down into pyruvic acid. This pyruvic acid is then converted to CO 2, ethanol,
and energy for the cell. The by-product, CO2, bubbles through the liquid and dissipates
into the air. The other by-product alcohol, remains in the liquid which is great for
brewers but not for the yeast because yeast dies and fermentation stops when the yeast
becomes poisoned by its own waste the alcohol.

2.1.7-1 Ethanol

Ethanol is a chemical compound having the formula C2H6O. Ethanol is

flammable with a boiling point of approximately 78 C. It is completely miscible
with water. It is a volatile, colorless liquid with a strong odor, and it tends to burn
with a smokeless blue flame which is not always visible in normal light. Ethanol
has a characteristic azeotrope with water. When mixtures of ethanol and water are
distilled at atmospheric pressure, the maximum concentration of ethanol in the
overhead distillate is approximately 96% by volume, with the remaining 4% by
volume of the overhead distillate being water. Ethanol has a specific gravity of
approximately 0.789 grams per cubic centimeter. (Grolier, 2001)

Ethanol is frequently produced industrially by fermentation processes. For

example, a source of organic material, such as a food source, can be inoculated with
yeast. The yeast will then begin to consume the organic material, and give off
ethanol as a metabolic waste product. Ethanol produced by micro-organisms tends
to be toxic to the micro-organisms, so when ethanol is produced above certain
levels it inhibits the micro-organisms and stops the fermentation process (Brad Cort,
2010). Micro-organisms are more efficient at converting some sorts of organic

16
compounds than others. The commonly used baker's yeast tends to be more efficient
at converting sugars in such things as sugar cane or corn than in converting
cellulosic materials into ethanol. Most ethanol production in the world now is based
on organic materials with high sugar content, such as corn or sugar cane. (Yah et. al,
2010)

After the fermentation process, the ethanol is typically recovered from the
fermentation mash. This is frequently done through distillation, and the ethanol is
recovered with water as an azeotrope. There are also ways to recover pure ethanol
from an azeotropic mixture of water and ethanol. For example, the collected
overhead ethanol and water can be run through a carbon absorption system to
absorb the water, leaving essentially pure ethanol.

The fermentation of an organic material with microorganisms is typically

done anaerobically, or not in the presence of oxygen. When little or no oxygen is
present, the micro-organisms find energy from the food source and not from
available free oxygen. Some yeast will preferentially produce water instead of
ethanol as a by-product of metabolism when oxygen is present. (Flickinger, 1999)

The yield of ethanol from agricultural crops can be estimated if the amount
of fermentable components sugar, starch, and cellulose is known prior to
fermentation. If the yield is predicted based on percentages at the time of harvest,
then the loss of fermentable solids during storage must be taken to account.
(Dawson and Boopathy, 2009)

Ethanol has a variety of uses in different industries. This chemical ranks

second to water as the most widely used solvent in chemical industry and as these
industries have expanded, so the demand for industrial alcohol has increased.
Alcohol acts as a solvent for an immense range of industrial products, including
paints, lacquers, dyes and oils. In addition, some are used as a raw material in
chemical synthesis and a little in the form of fuel (Ocloo and Ayernor, 2010). Fuels
available in the Philippines include ethanol to improve the efficiency of the engine
because of its anti-knock properties. (Eggeman, 2010)

17
 2.1.7-2 Carbon Dioxide

In the fermentation process, carbon dioxide is generated as a by-product,

and this carbon dioxide typically bubbles out of the fermentation mash. Carbon
dioxide is the harmless gas given off during fermentation and that responsible for
the bubbles in all fizzy drinks, including sparkling and slightly gassy wine. During
fermentation, pure carbon dioxide is given off for a considerable period of time.
The fermentation of six-carbon sugars by yeast results in the formation of carbon
dioxide as well as ethanol. For every 1 gallon of ethanol produced, 6.33 pounds of
carbon dioxide are formed. This ratio is fixed; it is derived from the chemical
equation:

C6H12O6 2C2H5OH + 2CO2 + heat

2.1.7-3 Other Co-products

The conversion and fermentation of agricultural crops yield products in

addition to ethanol and carbon dioxide. For example, even if pure glucose is
fermented, some yeast will be grown, and they would represent a co-product. These
co-products have considerable economic value, but, since they are excellent
cultures for microbial contaminants, they may represent a pollutant if dumped onto
land. Therefore, it becomes doubly important that these co-products be put to good
use.

2.1.8 Statistical Tests

2.1.8-1 Confidence Interval

The range that shows within which the true treatment effect is likely to lie is
called the confidence interval. These confidence intervals provide different pieces
of information from that of hypothesis tests which produces decision about any
observed difference either statistically significant or statistically not significant.
Confidence intervals provide a range about the observed effect size constructed in
such a way that how likely it is that the true value is captured is known. It is a

18
range of values for a variable of interest constructed to include the true value of the
variable at a specified probability which is called the confidence level. It is
conventional to create confidence intervals at the 95% level which means 95% of
the time properly constructed confidence intervals should contain the true value of
the variable of interest.

The upper and lower bounds of the confidence interval which is called
confidence limits gives information on how big or small the true effect might
plausibly be. If it is narrow, any effects far from this range have been ruled out by
the study and if it is wide it must have captured a diverse range of effect sizes and
any estimates of effect size will be quite imprecise.(Davies et al. 2009)

If t/2 is the t value with degrees of freedom n-1 leaving an area of /2 to the
right; x and s are the mean and standard deviation of a random sample from a
normal population with unknown variance 2 Confidence Interval for (mean) and
(standard deviation for whole population) unknown are computed as:

s s
xt / 2 < < x +t / 2 (Walpole et al. 2007)
n n

2.1.8-2 Analysis of Variance (ANOVA)

Analysis of variance (ANOVA) is a very common procedure used to deal

with testing population means for experiments involving one factor with more than
two levels. An ANOVA estimates group means, assess magnitude of variation
attributable to specific sources, partitions the variation according to source,
extension of 2-sample t-test to multiple groups, population model and sample
model. One-way ANOVA is of one type which is characterized by selecting random
samples of size n from each k (level) populations. The k different populations are
classified on the basis of a single criterion like treatment or groups. In the analysis
of variance, it is assumed that the k populations are independent and normally
distributed with means 1, 2... k and common variance of 2. ANOVA determines
statistically whether or not a significant difference in means across groups exists.
The hypotheses to be tested are the null hypothesis which states that all means are

19
equal and the alternative hypothesis is that at least one of the means is different
from the others. A significant result (null hypothesis is rejected) is obtained if the F-
value is F;k-1;N-1 and not to reject null hypothesis if otherwise.

The formulas that are used are:

Sample Variance SS ( of squares)

of t h e squared deviations between eac h observationt h e overall mean:

SST

of squared deviations between eac h observation

SSW
t h e meanof t h e group w h ic h it belongs

of t h e squared deviations betweeneac h group

SSB
meant h e overamean , weig h ted by t h e sample eac h groupn group

MSW (wit h groups mean squarevariationwit hgroup)

MSB(between groups mean squarevariation between group)

T h e ANOVA Table for k groupsN populations

Source of Sum of Squares Df Mean Square F
Variation MSB
Between Groups SSB k-1 MSB
MSW
Within Groups SSW N-k MSW

20
 Total N-1
2.1.8-3 Tukeys Test

To determine where among the individual means of the factors differ

significantly from the other, multiple comparison test such as Tukey test with equal
sample sizes of the different fermentation time and varying amount of yeast. Tukey
test tests the two tailed null hypothesis, H0: a = b where a and b represent all
possible combinations of the sample means in the factor. The first step in doing the
Tukey test is to calculate for the standard error which is given by the formula:

MS wit h
nobservation for equal sample sizes
s E=

The next step is to rank the sample means from lowest to highest for fermentation
time and another for the amount of yeast then obtain the difference of the paired
means within the factors. Pairing starts from the highest of the sample means
against the next smallest mean to the smallest. The difference is then compared to
Tukeys critical value multiplied by the standard error (CD). There is a significant
difference if the difference of the means is larger than the CD value, if not the two
means are declared to have no significant difference. If any non-significant
differences are detected between two means, then no comparisons are made for
other means within this interval.

21
2.2 RELATED STUDIES

Countries depended on imported fossils fuels, like the Philippines, feel the pinch
caused by the rising cost of crude oil and petroleum excise duties. Add to that, 70% of
mineral oil deposits are located in politically unstable areas.

In Asia, the UN Food and Agriculture Organization study has found that cassava
can be cultivated as an ethanol feedstock, without disrupting food production. The study
concluded that cassava can be utilized as source of renewable energy. Since the
Philippines is a tropical country, which is suitable for cassava production, we can
maximize this resource to meet the growing demand for energy.

Many studies have been conducted regarding the production of ethanol from
agricultural crops. Sugarcane provides a promising future as a reliable source of biofuel,
however certain crops may also be considered since they produce a considerable amount
of this renewable energy. Cassava, which contains a high level of starch according to the
study conducted by Leen Kuiper from Ecofys Netherlands on 2007, is one of the best
fermentable substances for the production of ethanol. He also added that cassava can be
grown in any different kind of soils and can be planted in a tropical weather like the
Philippines.

Ethanol is traditionally produced from feedstock high in sugar and/or starch

content by means of fermentation. In 2006, Maria Yuzon conducted a study on the
preparation of alcohol from Kamansi seeds using acid hydrolysis process. The alcoholic
distillate obtained underwent Iodoform, Lucas, and Chromic Anhydride test. It was
proven that starch extracted from Kamansi seeds is a good source of ethyl alcohol. Also,
Jimmy Santos described the preparation of alcohol from acacia fruits using a fermenting
mixture containing Trichoderma Viride. He employed fermentation using enzymatic
process and obtained an average alcohol content of 47.89%.

A study conducted by Dawson and Boopathy in 2008 shows that sugarcane

bagasse can be a viable source of bioethanol. In their study, they demonstrate that cane

22
processed bagasse could be used to produce fuel grade ethanol without saccharification.
A chemical pretreatment process using alkaline peroxide and acid hydrolysis was applied
to remove lignin, which acts a physical barrier to cellulolytic enzymes. Yeast
Saccharomyces cerevisiae ATCC, strain 765, was used in the experiment. The pre-
treatment process effectively removed lignin. The results indicate that ethanol can be
made from the sugarcane bagasse.

In production of bioethanol, certain factors must be considered. Based on the

Online Journal of Biological Science released in March 2010, temperature must be
optimized for bioethanol production from corn cobs using mixed yeast strain. In this
study, they employed two methods of sugar extraction, one is through dilute sulfuric acid
hydrolysis and the other is enzymatic hydrolysis. As a result, acid hydrolysis of corn cobs
gave higher amount of sugars than enzymatic hydrolysis. In their study about the time
and temperature of fermentation, the results showed that optimal temperature and time
for sugar fermentation were approximately 25 oC and 50 h by two yeast strains (S.
cerevisiae and P. Stipitis) respectively. At 20 and 40oC, less bioethanol was produced.
Varying the temperature of the fermentation process improves the effective utilization of
corn cobs sugar for bioethanol production can be achieved.

Alcohol produced from cassava flour hydrolysate (CFH) using rice malt as a
source of amylase for enzymatic hydrolysis of starch with standard glucose and sucrose
used as controls is the focus of the study of Ocloo and Ayernor from Journal of Brewing
and Distilling dated last year 2010. The conversion efficiency of sugar to alcohol, rate of
fermentation and types of alcohol produced were determined. The effect of yeast
concentrations using the combined strain of yeast (A. niger and Z. mobilis) and
fermentation time (24, 36, 48, and 60 h) on the yield of alcohol from CFH were also
studied. Results showed that the conversion efficiency of sugars to alcohol was 99.51%
and 95.37% for cassava flour hydrolysate (CFH), standard glucose and sucrose solutions
respectively. It is therefore concluded that using glucose solution yielded more alcohol
than alcohol produced using standard sucrose solution. Alcohol produced was mostly
ethanol with traces of methanol. The study suggests that high yield of alcohol could be
produced from cassava flour hydrolysate.

23
 Generally, bioethanol from corncobs are produced using simultaneous
saccharification and fermentation with the aid of Aspergillus niger and Saccharomyces
cerevisae. This is the center of the study presented by F.S. Johnson in April of 2009. He
pointed out that maize is the most abundant cereal grown in many countries and is
accompanied by enormous amount of agro wastes of which corncobs form 30%. This
agro waste which is currently under utilized was used to produce bio-ethanol. Aspergillus
niger isolated from soil sampled was used to hydrolyze the corncobs into simple sugars.
Filtrate obtained from corncobs broth fermented by A. niger was used as crude enzyme in
optimization tests on corncobs powder suspended in 50 mM citrate buffer pH 5.0.
Optimum temperature, pH and substrate concentration for saccharification were 40oC, 4.0
and 6% respectively. Saccharomyces cerevisae was added to A. niger filtrate to cause
fermentation of the corncobs. By applying this method, he concluded that the highest
ethanol concentration of 0.64 g/l was recorded over the 24 h fermentation period.

Yeast is an important ingredient in fermentation. Zhang and Feng conducted a

research regarding the fermentation potential of Zymomonas mobilis in ethanol
production from low-cost raw sweet potato. The effects of pH, high concentration of
glucose and the initial ethanol content on the fermentation process of ethanol with three
strains of Zymomonas mobilis were investigated and the strain of ATCC. It was found that
the condition for ethanol production was optimized to be pH 4, substrate concentration of
20%, and time of fermentation of 24 h, resulting in ethanol yield of 66.4 g/L and
fermentation efficiency of 93.5%, respectively. This work demonstrates that the low-cost
sweet potato is a feasible feedstock for ethanol fermentation with Z. mobilis.

The Department of Agricultural Engineering led by B. A. Adelekan investigates

on ethanol productivity of cassava crop as a sustainable source of biofuel in tropical
countries. This study found that a total of 6.77 million tonnes or 1338.77 million gallons
of ethanol are available from total cassava production from tropical countries. The
production and use of ethanol from cassava crop is recommended in the cassava-growing
tropical countries of the world like the Philippines.

24
 In 2004, the Canadian Firm Iogen brought the first cellulose-based ethanol plant
on-stream. The primary consumer thus far has been the Canadian Government, which
along with the United States Government particularly the Department of Energys
National Renewable Energy Laboratory, has invested millions of dollars into assisting the
commercialization of cellulosic ethanol. Realization of this technology would turn a
number of cellulose-containing agricultural by-products, such as corn cobs, sugarcane
and wheat, into renewable energy resources.

Philippines has a huge potential of utilizing cassava as a feedstock for the

production of ethanol since its climate is suitable for growing a massive volume of this
root crop. The Department of Agricultural Economics in UPLB released a paper that
discusses the potential of cassava as a source of feedstock for bioethanol and the major
policy considerations for developing the industry. The paper provides some
recommendations on enhancing the viability of cassava as a source of bioethanol in the
Philippines. However as the paper explains, there are certain important concerns that
need to be addressed for the project to be viable. These include the market, productivity
of cassava production, cost of cassava feedstock with other feedstock sources, potential
production areas, production technologies, feedstock supply arrangements between
feedstock producers and processing plants, incentives for industry players, and impact on
the environment.

These related studies proved that ethanol can be obtained from a different variety
of agricultural crops with a considerable yield which can be optimized to be the source of
renewable energy and help sustain the growing demand for energy.

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