Chapter I

The Problem and Its Setting

In this chapter represents the background information of the study, the problem

encountered towards the study, significance of the study, and the limits towards the

study.

Background of the Study

Increase in the population over the last century lead to the increase of the energy

consumption worldwide. The expansion of petroleum products joined with consumption

of these stored mineral oil prompted the advancement of eco-accommodating ideas.

Everywhere throughout the world, governments have supported the use of alternative

sources of energy for dominating energy crisis. The higher cost of oil has pulled the

greater attention to biofuels, especially bioethanol. Moreover, it was stressed out that

crude oil is no longer considered as renewable energy.

In spite of this, some businesses interest and research proceeds with in light of

the abundance of crude materials and the prediction that the energy economics will

change the not so distant future to support bioethanol. Bioethanol is a promising

sustainable power source, eco-accommodating and causes most extreme decrease of

negative natural effects produced by the overall use of fossil fuels.

In this context, the researchers focuses on the extraction of bioethanol on

marang Seeds.
Statement of the Problem

The study was conducted to extract bioethanol from marang seeds.

Specifically, it aimed to provide answers to the following questions:

1. What is the percentage of the alcohol content produced from each sample?

1.1 S1 (Control) = Distilled water + 20g Muscuvado + 20g commercial

yeast

1.2 S2= 400 ml of hydrolyzed mixture of Marang (Artocarpus

odoratissimus) seeds + 20g Muscuvado + 10g commercial yeast

1.3 S3= 400 ml of hydrolyzed mixture of Marang (Artocarpus

odoratissimus) seeds + 20g Muscuvado + 20g commercial yeast

1.4 S4= 400 ml of hydrolyzed mixture of Marang (Artocarpus

odoratissimus) seeds + 20g Muscuvado + 30g commercial yeast

2. What are the significant differences between the control and the following

samples?

2.1 S1 (Control) = Distilled water + 20g Muscuvado + 20g commercial

yeast

2.2 S2= 400 ml of hydrolyzed mixture of Marang (Artocarpus

odoratissimus) seeds + 20g Muscuvado + 10g commercial yeast

 2.3 S3= 400 ml of hydrolyzed mixture of Marang (Artocarpus

odoratissimus) seeds + 20g Muscuvado + 20g commercial yeast

2.4 S4= 400 ml of hydrolyzed mixture of Marang (Artocarpus

odoratissimus) seeds + 20g Muscuvado + 30g commercial yeast

3. At what sample does the wasted Marang seeds produced the highest

percentage of alcohol content.

3.1 S1 (Control) = Distilled water + 20g Muscuvado + 20g commercial

yeast

3.2 S2= 400 ml of hydrolysed mixture of Marang (Artocarpus

odoratissimus) seeds + 20g Muscuvado + 10g commercial yeast

3.3 S3= 400 ml of hydrolyzed mixture of Marang (Artocarpus

odoratissimus) seeds + 20g Muscuvado + 20g commercial yeast

3.4 S4= 400 ml of hydrolyzed mixture of Marang (Artocarpus

odoratissimus) seeds + 20g Muscuvado + 30g commercial yeast

4. At what sample does the hydrolysed marang seeds shows the greatest

potential in producing bioethanol?

4.1 S1 (Control) = Distilled water + 20g Muscuvado + 20g commercial

yeast

4.2 S2= 400 ml of hydrolysed mixture of Marang (Artocarpus

odoratissimus) seeds + 20g Muscuvado + 10g commercial yeast

 4.3 S3= 400 ml of hydrolyzed mixture of Marang (Artocarpus

odoratissimus) seeds + 20g Muscuvado + 20g commercial yeast

4.4 S4= 400 ml of hydrolyzed mixture of Marang (Artocarpus

odoratissimus) seeds + 20g Muscuvado + 30g commercial yeast

Hypotheses

Null

The produced bioethanol from marang seeds has low percentage of alcohol.

Alternative

The produced bioethanol from marang seeds has very high percentage of alcohol.

Significance of the Study

The study aimed to produce bioethanol using wasted Marang (Artocarpus

odoratissimus) seeds by extracting the bioethanol from marang seeds and determining

its alcohol content. Also, the study was conducted to provide baseline information about

its significance in our society or community. To the community, the result of the study

helped to add the economic value of the marang seeds and to find an alternative source

of bioethanol which is from the wasted marang seeds.

Scope and Limitation

 The study was limited only on the production of ethanol from marang (Artocarpus

odoratissimus) seeds through the process of hydrolysis, fermentation and distillation.

And by determining its alcohol content. The study used the process of distillation after

the hydrolysis and fermentation subjecting the mixture into four trials S 1 (Control) =

Distilled water + 20g Muscuvado + 20g commercial yeast, S 2= 400 ml of hydrolyzed

mixture of Marang (Artocarpus odoratissimus) seeds + 20g Muscuvado + 10g

commercial yeast ,S3= 400 ml of hydrolyzed mixture of Marang (Artocarpus

odoratissimus) seeds + 20g Muscuvado + 20g commercial yeast, S4= 400 ml of

hydrolyzed mixture of Marang (Artocarpus odoratissimus) seeds + 20g Muscuvado +

30g commercial yeast

The study was limited only on the percentage of the alcohol present in each

sample.

Operational Definition of Terms

The definition of terms is for the benefit of the reader’s understanding. The terms

are defined based on how it was used in the study.

Amylase- any member of a class of enzymes that catalyse the hydrolysis of starch into

smaller carbohydrate molecules such as maltose.

Bioethanol- an ethanol derived exclusively from the fermentation of plant starches. The

main goal of the study is to produce bioethanol.

Distillation- the process of separating liquid. The objective of the study is to separate

the liquids based on the differences of their volatility.

Fermentation- the breakdown of pyruvic acid without the use of oxygen also known as

anaerobic respiration.

Hydrolysis- a process defined as the chemical reaction where something reacts with

water and is changed into new substance. Hydrolysis is used for water separation.

Starch- a carbohydrate that is the chief form of stored energy in plants, especially

wheat, corn, rice, and potatoes mixture of two different polysaccharides built out of

glucose units, and forms a white, tasteless powder when purified.

Saccharification- the process of breaking a complex carbohydrate (as starch) into

simple sugars.

Sugar- a sweet crystalline substance obtained from various plants, especially sugar

cane and sugar beet, consisting essentially of sucrose, and used as a sweetener in food

and drink.

Yeast - enzymes chemically breakdown the sugars into products that the cell can used.

Yeast is used to manufacture ethanol through fermentation.

 Chapter II

Review of Related Literature

This chapter presents related studies of the problem in hand. This gives

additional background information to support the study. It tackles about the potential of

wasted Marang (Artocarpus odoratissimus) seeds extract as a source of bioethanol.

Marang (Artocarpus odoratissimus)

Artocarpus odoratissimus, otherwise known as Marang fruit, is a rich source of

energy and contains tidy sum of saccharide eventually broken into glucose. Packed with

protein and fat, it is also known as athlete’s fruit and are very much compatible to the

banana. Moreover they are also rich in protein, fat, sugar, carbohydrates, ash, calcium,

phosphorous, smoothing iron, crude fiber, retinol, Beta -carotene, nicotinic acid,

thiamine, lactoflavin, and Vitamin A and C. Marang fruit is a tree in the mulberry and fig

family Moraceae. It is native to Borneo, Palawan, and Mindanao Island, and is closely

related to the jackfruit, cempedak, and breadfruit trees which all belong to the same
genus, Artocarpus odoratissimus. It grows to 25 meters (82 ft.) tall. The leaves are 16–

50 cm long and 11–28 cm broad, similar to the Breadfruit's, but are a little less lobed.

Many trees lose the leaf lobbing once mature. The appearance of the fruit can be

regarded as an intermediate shape between the jackfruit and the breadfruit. It is round

to oblong, 15–20 cm long and 13 cm broad and weighing about 1 kg. The thick rind is

covered with soft, broad spines. They become hard and brittle as the fruit matures.

When fully mature the expanding arils stretch the outer rind which often appears lumpy,

especially if not all seeds was pollinated (Fahad, 2015).

Bioethanol

Bioethanol is used in fuel industry as an additive for petrol. The most common

blend in 90% petrol and 10% ethanol. Bioethanol is completely comprised of biological

products, such as the combustion of bioethanol results in cleaner combustion (CO2,

steam and heat). Carbon dioxide is absorbed by plants through the process of

photosynthesis to help the plant grow. This cycle of reaction and energy combustion

means bioethanol could potentially be a carbon neutral fuel source (Pirolini, 2015).

Ethanol production from lignocellulosic biomass follows the process of (a)

hydrolysis of cellulose and hemicellulose; (b) sugar fermentation; (c) separation of lignin

residue and, (d) recovery and purifying the ethanol to meet fuel specifications. They

added that the task of hydrolyzing lignocellulose to fermentable monosaccharides is still

technically problematic because the digestibility of cellulose is hindered by many

physico-chemical, structural and compositional factors. Owing to these structural

characteristics, pretreatment is an essential step for obtaining potentially fermentable

sugars in the hydrolysis step. They explained that the purpose of the pretreatment is to

break down the lignin structure and disrupt the crystalline structure of cellulose for

enhancing enzymes accessibility to the cellulose during hydrolysis step (Mosier ET. Al,

2005).

Put simply, bioethanol is energy made from living matter, usually plants.

Bioethanol, biodiesel, and biogas are types of biofuels. Bioethanol are considered

renewable energies, emit less than fossil fuels, and have received increasing attention

in the transition to a low-carbon economy.

Ethanol is the most generally utilized fluid biofuel. It is a liquor and is matured

from sugars, starches or from cellulosic biomass. Cellulosic materials can be utilized to

create bioethanol. Bioethanol speaks to a critical, sustainable fluid fuel for engine

vehicles. Creation of bioethanol from biomass is one approach to lessen both the

utilization of unrefined petroleum and natural contamination. Change advances for

delivering ethanol from cellulosic biomass assets, for example, woodland materials,

agrarian build-ups and urban squanders are being worked on and have not yet been

shown economically. Keeping in mind the end goal to create bioethanol from cellulosic

biomass, a pre-treatment procedure is utilized to decrease the example measure,

separate the hemicelluloses to sugars, and open up the structure of the cellulose

segment. The cellulose partition is hydrolyzed by acids or compounds into glucose

sugar that is aged to bioethanol. The sugars from the hemicelluloses are likewise aged

to bioethanol. The utilization of bioethanol as an engine fuel has as long a history as the

auto itself. It started with the utilization of ethanol in the interior ignition motor (Saini,

2014)
Enzymatic Hydrolysis

Grohmann and Bothast (1997) examined a consecutive saccharification of

polysaccharides in corn fiber by treatment with weaken sulfuric corrosive at 100– 160

°C took after by incomplete balance and enzymatic hydrolysis with blended cellulase

and amyloglucosidase chemicals at 45 °C. The successive treatment accomplished

high (roughly 85%) change of all polysaccharides in the corn fiber. In any case, the

arrangement of mixes inhibitory to fermentative microorganisms turned out to be

promptly clear at all pretreatments tried at 140 and 160 °C.

The effect of the enlightenment with white straightly enraptured light (WLPL) of

two financially accessible cellulases from Trichoderma reesei on their movement in

hydrolysis of microcrystalline cellulose was examined. Chemicals were enlightened with

WLPL for 60 min and 120 min and for every local and lit up protein test particular

movement and energy of catalyst catalyzed hydrolysis of microcrystalline cellulose were

built up. Sub-atomic weight Mw and radii if gyration Rg of protein chains of local and lit

up compounds were estimated by methods for high weight measure exclusion

chromatography combined with multiangle laser light disseminating and refractometric

identifiers (HPSEC-Shopping centers RI). Compliances of protein chains of local and lit

up compounds were assessed based on their round dichroism (Album) spectra. Also,

sub-atomic weight Mw and radii of gyration Rg of polysaccharide chains of

microcrystalline cellulose local and processed for 10 min, 480 min and 1440 min with

unique and WLPL invigorated proteins WT and TR were taken. Light with WLPL of both

cellulases examined did not change auxiliary structures of protein particles of local

chemical. Atomic weight Mw and radii of gyration Rg of enlightened chemicals varied

extraordinarily from those found for local compounds. Enlightenment of compounds

prompted increment of particular action and rate constants of response of hydrolysis

microcrystalline cellulose catalyzed by lit up proteins as contrasted and local chemicals

(Polit et.al, 2012).

Saccharification

The hydrolysis of polysaccharides to solvent sugars is called "saccharification".

Malt produced using grain is utilized as a wellspring of β-amylase to separate starch into

the disaccharide maltose, which can be utilized by yeast to create brew. Other amylase

chemicals may change over starch to glucose or to oligosaccharides.

Making sugar from the starch saves. In this administration the diastatic chemicals

begin following up on the starches, separating them into sugars. The amylases are

compounds that work by hydrolyzing the straight chain bonds between the individual

glucose atoms that make up the starch chain. A solitary straight chain starch is called

an amylose. An extended starch chain is called an amylopectin. These starches are

polar particles and have distinctive finishes. An amylopectin contrasts from an amylose

by having an alternate sort of atomic bond at the branch point, which isn't influenced by

the diastatic catalysts (John, 2015).

Ordinary technique utilized for the generation of bioethanol from cassava starch

for the most part requires the fundamental gelatinization step took after by liquefaction

and saccharification. The sugar shaped amid these procedures is additionally matured

utilizing either yeast or microscopic organisms. Since, starch got from any plant source

is a perplexing atom, it require different hydrolytic chemicals for its transformation into
basic fermentable sugars. Among numerous extracellular hydrolases accessible,

microbial amylases are as often as possible utilized for its change. For business

creation of amylases Aspergillus and Rhizopus species are viewed as most critical

sources on the grounds that the proteins from these sources are by and large the most

able and are accessible in extreme amounts. Despite several advantages of

simultaneous saccharification and fermentation using multiple organisms, there are also

few shortcomings (Sidra et.al, 2014).

Ethanol Fermentation

Sugar from plant material is converted into ethanol and carbon dioxide by

fermentation. The enzymes found in single-celled fungi (yeast) are the natural catalysts

that can make this process happen (Bitesize, 2014).

Ethanol (C2H5OH) is a light alcohol and is a volatile, colourless, flammable liquid

with a characteristic odour. It is also known as ethyl alcohol and often abbreviated as

EtOH. The most common way of production is the fermentation of sugar or starch from

agricultural crops by yeasts or bacteria. Ethanol can either be used as high blend (E85)

in dedicated flex-fuel vehicles, or as low blend in most current vehicles without

modifications. The blending rate is usually up to 10%.The feedstock for ethanol

production can be any material containing appreciable amounts of sugar or substances

that can be converted to sugar. Conventional production uses sugar, starch or other

polysaccharides. The production process of second generation ethanol, also called

cellulosic alcohol, uses cellulosic feed stock which require further pretreatment. In a

fermentation process sugar is converted to ethanol by microbes, which are inoculated to

the feedstock. The monosachharides originate either directly from disaccharides, which

are broken up via invertase enzymes, or from starch which is hydrolysed with amylase

enzymes. In addition to ethanol, water and carbon dioxide are produced also

(Bioenergy, 2014).

Ethanol Production

The Philippines as of late documented a yearly biofuels report with the USDA

Outside Agrarian Administration's Worldwide Farming Data System, taking note of

ethanol generation is relied upon to increment through one year from now because of

the development of limit. As indicated by the report, there were eight ethanol plants

working in the Philippines a year ago with a consolidated limit of 222 million liters (58.65

million gallons). The country created 168 million liters of ethanol a year ago, up 46

percent when contrasted with the 115 million liters delivered in 2014. Sugarcane and

molasses are as of now utilized as ethanol feedstocks in the nation. Two extra ethanol

plants started activities not long ago, including one recently built ethanol plant and a

consumable liquor maker that started delivering fuel ethanol. Together, the offices have

a yearly limit of 60 million liters. This year, ethanol generation is again anticipated that

would develop, achieving 266 million liters, up roughly 58 percent when contrasted with

2015. Limit use is required to reach 94 percent this year, up 76 percent from a year ago.

In 2017, another ethanol plant is relied upon to start activities, bringing the aggregate

number of ethanol plants to 11. The extra 40 million liters of limit is relied upon to build

by and large limit in the nation to 322 million liters. The Philippines government at
present expects ethanol to contain 10 percent of the aggregate yearly volume of fuel

disseminated by oil organizations in the nation. The nation at present means to build the

mix level 20 percent in 2020 and 85 percent in 2025. The report likewise shows the

ethanol imports in the Philippines fell a year ago, basically because of expanded

neighbourhood generation. Fuel ethanol imports, which achieved 339 million liters in

2014, fell 8 percent a year ago, achieving 311 million liters. Imports from the U.S. fell by

an expected 28 percent, from 246 million liters in 2014 to 178 million liters in 2015. The

U.S., be that as it may, in any case represented 57 percent of aggregate fuel ethanol

imports a year ago, down from 73 percent in 2014 (Erin,2016).