Production of Bioethanol from Saba banana

(Musa acuminata x Musa balbasiana) peel

by Separate Hydrolysis and Fermentation
using Saccharomyces cerevisiae and Benzathine penicillin g.
• Introduction
• Statement of the problem
• Significance of the study
• Scope and Delimitation
• General Objective
• Specific Objective
• Related Literature
• Methodology
• Gannt Chart
 Bioethanol
Banana Peel Waste
Statement of the Problem
Sources of municipal solid waste in the Philippines, 2008-2013
Source: (http://nswmc.emb.gov.ph)
Composition of municipal solid waste in the Philippines, 2008-2013
Source:(http://nswmc.emb.gov.ph)
 Bioethanol
Banana Peel Waste

•lessen amount of waste

•reduce carbon

Significance of the study

 Scope and Delimitations
The experimentation and process deals:

• production of bioethanol out of fermented saba banana peelings.

• the acid that will use for pretreatment process will be 0.5% (v/v)
sulfuric acid.
• The conversion of cellulose to fermentable sugars is through
enzymatic hydrolysis. The use of Trichoderma reesei:Aspergilus
niger with hydrolysis time of 24hr, 48hr, and 72hr and with a ratio
of 2:1 at 35oC.
• The amount of sugars is determined by Benedict’s test.
• The fermentation process will do by pilot scale measurement with
the use of Saccharomyces cerevisiae only and addition of
Benzathine penicillin g.
• The bioethanol will be purify by simple distillation
• characterized by GC-FID.
General Objectives

To produce bioethanol from saba banana (Musa

acuminata x Musa balbasiana) peel as an
alternative source of bioethanol via separate
hydrolysis and fermentation process.
 Specific Objectives
• Evaluate the effect of treated (with sulphuric acid) and untreated
saba banana peel on the amount of sugar produced.
• To determine the effect from 24hr, 48hr, 72hr hydrolysis time at the
2:1 ratio Trichoderma reesei:Aspergilus niger.
• To determine the amount of sugars formed prior to fermentation
during enzymatic hydrolysis time of 24hr, 48hr, and 72hr using
Benedict’s solution.
• To determine bioethanol yield by Saccharomyces cerevisiae in
fermentation
• To determine bioethanol yield by combining Saccharomyces
cerevisiae and addition of Benzathine penicillin g. in fermentation.
• To determine the ethanol concentration using GC-FID.
CHAPTER 2
REVIEW OF
RELATED
LITERATURE
WORLD EMISSION of CO2
• 1850: United Kingdom was a top emitter of CO2
nearly six times higher than United States
• 2011: Countries in Asia
• contributes CO2
Greenhouse effect:
1880 to 2013:
0.8 oC to 14.6oC
SHIFTING to RENEWABLE RESOURCES

• CO2 produced from biomass was absorbed by the

atmosphere by photosynthesis in plants.
 TOP BIOETHANOL PRODUCER in the
WORLD
• Bioethanol from Corn in US
• Bioethanol from Sugarcane in Brazil
• Bioethanol from Sugarbeet juice in European Countries
• Bioethanol from Cassava, Sweet Sorgum, and Sweet Potato in China

World Fuel Ethanol Production by Country or Region (Million Gallons)

Country 2007 2008 2009 2010 2011 2012 2013 2014 2015

USA 6,521 9,309 10,938 13,298 13,948 13,300 13,300 14,300 14,806

Brazil 5,019 6,472 6,578 6,922 5,573 5,577 6,267 6,190 7,093

Europe 570 734 1,040 1,209 1,168 1,179 1,371 1,445 1,387

China 486 502 542 542 555 555 696 635 813

Canada 211 238 291 357 462 449 523 510 436
Rest of
World 315 389 914 985 698 752 1,272 1,490 1,147

WORLD 13,123 17,644 20,303 23,311 22,404 21,812 23,429 24,570 25,682
 Banana for Bioethanol

According to Philippines Statistics Authority, “banana production went up by 2.6 percent from 2.05
million metric tons in 2016 to 2.10 million metric tons this year 2017” (www.psa.gov.ph), where this
increase in production of banana, saba banana have a 9 percent participation in 2.10 million metric
tons.
According to (Velásquez-Arredondo, Ruiz-Colorado, & De
Oliveira, 2010)

• Banana pulp: 53.2% (w/w) starch content, good

for acid hydrolysis
• Flower Stalk: 40.9% (w/w) cellulosic material
good for enzymatic hydrolysis
• Banana skin: Higher LHV: best raw material for
fuel
STAGES of BANANA
• Stage 1: 20 days
• Stage 2: 35 days
• Stage 3: 50 days
• Stage 4: 65 days
• Stage 5: 80 days
SABA BANANA (khawas et al)
PRE-TREATMENT
 ACIDIC PRE-TREATMENT

▫ Dilute Acid Pre-treatment

▫ Steam Explosion Pre-treatment
▫ Organosolv Pre-treatment
 Dilute Acid Pre-treatment
• Sulfuric acid
• Reduce lingocellulosic recalcitrance by
disrupting covalent bonds
• Reduce crystallinity of cellulose
• Increase porosity of material
• Makes accessible to cellulates during Enzymatic
hydrolysis
• Performed at 120oC-210oC
• pH 4.5-5.0
 Steam Explosion Pre-treatment
• Biomas are treated to high pressure to increase
chemical and biological activity
• Performed at 160-260oC
• Pressure of 0.69-4.83MPa
• Sulfuric Acid as catalyst
 Organosolv Pre-treatment
• Partially hydrolysed
• Hemicellulose is hydrolysed into soluble
components such as oligosaccharides,
monosaccharides and acetic acid.
 ENZYMATIC HYDROLYSIS
• Helps breakdown polysaccharides into
monomers for fermentation and distillation
• Reacts directly with cellulose using
enzymes(cellulase)
• Cellulase enzyme can be derived from bacteria
or fungi
• Variety of Species (Tricchoderma, Penicillum,
Aspergillus)
• Performed at 50oC, pH 5.0, 64hr
 Trichoderma reesei
• has the capacity to secrete large amounts of
cellulolytic enzymes (cellulases and
hemicellulases)

• According to (nuniek Hendrianie, 2015)

Trichoderma reesei produces endoglukanase
and eksoglukanase to 80% but lower β-
glukosidase
 Aspergillus niger
• According to (Philomena Kanwulia Igbokwe, April
2016) Aspergillus niger to change the content of
cellulose into glucose in the banana peel.

• According to (nuniek Hendrianie, 2015) these

microorganisms produce high β-glucosidase but low
endo-β-1.4-glucanase and exo-β-1,4-glukanasenya

• pH 4.5-5.5
 Fermentation
• Fermentation process was the chemical
breakdown of a substance by bacteria, yeasts or
any other microorganism.

• Saccharomyces cerevisiae
• Benzathine penicillin g.
Distillation
• Distillation process is the action of purifying a
liquid by a process of evaporation and
condensation. The liquid ethanol are being
heated at the 78oC to separate in water. The
ethanol is vaporize and being cooled in
condenser making it liquid and collected in flask
 Chapter 3

Methodology
 SAMPLE PRE-TREATMENT ENZYMATIC HYDROLYSIS

DISTILLATION FERMENTATION TEST FOR REDUCING SUGAR

TEST FOR ETHANOL

CONCENTRATION

Process Flow Chart of Production

Bioethanol from Saba Banana Peels
 Materials
> Major >Minor

•Saccharomyces cerevisiae

•Benzathine penicillin g.

•Aspergillus niger

•Tricchorderma reesei
 Reagents

• Sulfuric Acid
• Anhydrous sodium carbanate
• Copper(II) sulfate pentahydrate
• Sodium citrate
• Citric Acid
 Equipment

Pressure Cooker . Analytical Balance .

Simple distillation setup .Gas Chromatography
 Glasswares

Beaker . Graduated Cylinder . Erlenmeyer Flask . Volumetric Flask .

Reagent Bottle . Pipette . Stirring rod . Funnel . Watch glass
Experimental
Procedure
Preparation of Raw Material
sample collect in
Tenement Market

cut into tubes

wash with
distilled water

dry in sunlight
(48hrs)

dry in oven @ 65oC

grind
Dilute Acid Pre-treatment
Sample A Sample B
(500g) (500g)

0.5 Sulfuric Acid

Autoclave at 120-210°C
for 30mins

cool @ room temp.

filter

wash saba banana peel

soak in citrate buffer

enzymatic hydrolysis
Enzymatic Hydrolysis
Sample A Sample B
(100g) (100g)

200ml of citrate buffer

2.4g cellulose

stand for 24, 48 and 72 hours

filter

characterization of reducing sugar

Characterization of Reducing Sugar
Sample A Sample B
(1ml) (1ml)

2ml Benedict’s solution

water bath for 3-5mins

observe color change

fermentation
 Fermentation
Sample A1 Sample A2
(250g) (250g)

0.5:0.05g
0.5g Saccharomyces
Saccharomyces cerivisiae cerivisiae:Benzathine
penicilline g.

room temp 35°C

ferment for 9 days

distillation
 Distillation
Sample A1 Sample A2
(500ml) (500ml)

heat at 78°C

vaporize

cool in condenser

characterization of ethanol
 Thesis Gantt Chart Activities
Thesis Gaant Chart September October November
Activity Description W W W W W W W W W W W W
No. 1 2 3 4 5 6 7 8 9 10 11 12
1 Collection and Purchasing
- Purchasing Tricchoderma reesei, Aspergillus
niger, Saccharomyces c. and Benzathine
penicillin g.
- Purchasing chemicals needed (Sulfuric acid,
Copper Sulphate, Sodium Carbonate, Sodium
Citrate)
- Purchasing materials (Pressure cooker, 2L
Beaker, Buchner funnel)
- Collecting Saba Banana peel
2 Production of bioethanol
- Preparation of cellulase
- Sample Preparation (Drying and crushing of
saba banana peel)
- Dilute Acid Pretreatment
- Enzymatic Hydrolysis
- Characterization of reducing sugar
- Fermentation
- Distillation
3 Characterization of Bioethanol (GC-FID)
4 Evaluation of results
5 Final report

General Activity Detailed Task per Activity

 Thesis Gaant Chart December January
Activity Description W W W W W W W W
No. 13 14 15 16 17 18 19 20

1 Collection and Purchasing

- Purchasing Tricchoderma reesei, Aspergillus niger,
Saccharomyces c. and Benzathine penicillin g.
- Purchasing chemicals needed (Sulfuric acid, Copper Sulphate,
Sodium Carbonate, Sodium Citrate)
- Purchasing materials (Pressure cooker, 2L Beaker, Buchner
funnel)
- Collecting Saba Banana peel
2 Production of bioethanol
- Preparation of cellulase
- Sample Preparation (Drying and crushing of green saba banana
peel)
- Dilute Acid Pretreatment
- Enzymatic Hydrolysis
- Characterization of reducing sugar
- Fermentation
- Distillation
3 Characterization of Bioethanol (GC-FID)
4 Evaluation of results
5 Final report

General Activity Detailed Task per Activity

THANK YOU! 