How to cite:
Rizka Lestari Dewi and Anggun Puspitarini Siswanto. (2021). Utilization of
bioetanol fermentation waste pineapple and coconut water as disinfectants
with bacteria saccharomyces cerevisiae. Journal Eduvest. 1(6): 423-436
E-ISSN:
2775-3727
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https://greenvest.co.id/
Eduvest Journal of Universal Studies
Volume 1 Number 6, June 2021
p- ISSN 2775-3735- e-ISSN 2775-3727
UTILIZATION OF BIOETANOL FERMENTATION WASTE PINEAPPLE AND
COCONUT WATER AS DISINFECTANTS WITH BACTERIA
SACCHAROMYCES CEREVISIAE
Rizka Lestari Dewi and Anggun Puspitarini Siswanto
Diponegoro University
ARTICLE INFO ABSTRACT
Received:
May, 25
th
2021
Revised:
June, 4
th
2021
Approved:
June, 15
th
2021
Research analysis of sugar content and the effect of
ethanol content on bioethanol from old coconut water and
pineapple peel with the help of Saccharomyces
Cerevisease bacteria. The condition of the spread of the
Corona Virus or COVID-19 in Indonesia, thus making
bioethanol produced from fermenting pineapple peel
waste and old coconut water for disinfectant products to
spray around homes and public places to reduce bacteria
and viruses. The production of bioethanol is carried out by
pre-treating coconut water and pineapple peel, the
fermentation stage with Saccharomyces cerevisiae yeast
and the distillation stage. The result of the highest
bioethanol content was 32% with a mass of 5 g yeast with
a time of 24 hours. The highest calorific value at 72 hours
was 211.95 kcal/kg. The result of the highest specific
gravity at 24 hours and the mass of yeast 4 g is 0.98 g/ml.
Based on the bioethanol quality requirements, the
bioethanol produced is not in accordance with the
bioethanol quality requirements, this is due to the absence
of nutrient decomposing bacteria so that it is less than
optimal in converting glucose into bioethanol.
KEYWORDS
Bioethanol, Disinfectant, Fermentation, Distillation
This work is licensed under a Creative Commons
Attribution-ShareAlike 4.0 International
Rizka Lestari Dewi and Anggun Puspitarini Siswanto
Utilization of bioetanol fermentation waste pineapple and coconut water as
disinfectants with bacteria saccharomyces cerevisiae 424
INTRODUCTION
In 2019, a new disease caused by a virus appeared and attacked the respiratory
system, this disease is called the Corona Virus or COVID-19 (Ramesh, Siddaiah, &
Joseph, 2020). The COVID-19 cases started with pneumonia or mysterious pneumonia.
Coronaviruses are a large family of viruses that cause illness ranging from mild to severe
symptoms (Putri, 2020). There are at least two types of coronavirus that are known to
cause diseases that can cause severe symptoms such as Middle East Respiratory
Syndrome (MERS) and Severe Acute Respiratory Syndrome (SARS) (Timah, 2021).
Coronavirus Disease 2019 (COVID-19) is a new type of disease that has never been
previously identified in humans (Bahtiar & Ariyanti, 2021).
Common signs and symptoms of COVID-19 infection include acute respiratory
symptoms such as fever, cough and shortness of breath. The average incubation period is
5-6 days with the longest incubation period of 14 days (Yuzar, 2020). Clinical signs and
symptoms reported in the majority of cases were fever, with some cases having difficulty
breathing, and X-rays showed extensive pneumonia infiltrates in both lungs (Nurhayatun
& Prabowo, 2020). Appropriate action has been taken to prevent and limit this spread to a
wider and faster pace (Nicola et al., 2020). Presence of inhibitory chemicals in
lignocellulose hydrolysates is a major hurdle for production of second-generation
bioethanol. Especially cheaper pre-treatment methods that ensure an economical viable
production process generate high levels of these inhibitory chemicals (Vanmarcke,
Demeke, Foulquié-Moreno, & Thevelein, 2021).
Bioethanol as renewable fuel addresses elevated production costs, as well as food
security concerns (Nwaefuna, Rumbold, Boekhout, & Zhou, 2021). The right precaution
is to use natural disinfectants from the manufacture of bioethanol produced by fermented
pineapple peel waste and old coconut water (Prasetyo, 2011). In bioethanol, this is useful
for a disinfectant mixture so as to take advantage of the surrounding waste (Novia,
Windarti, & Rosmawati, 2014). Using coconut water as bioethanol is one of the
ingredients for making bioethanol. In coconut water, natural chemical content is very
good for the body, including: Vitamin C which has a description of the content in it, such
as nicotinic acid, folic acid, pantothenic acid, bitin, and riboflavin (Andari, Mulyadi, &
Puspawiningtyas, 2015). Besides containing minerals, coconut water also contains sugar
in the range of 1.7-2.6%, 0.07-0.55% protein and contains potassium (Faizal, Zuhandri, &
Andrio, 2011).
In addition to the old coconut water, use of waste can be used as alternative fruit
is pineapple skin (Noviandi, Yaman, & Rinidar, 2018). The content of pineapple skin that
can be processed and used as a basic ingredient in the manufacture of bioethanol. The
content of pineapple skin contains carbohydrates by 10.54%, ash by 0.48%, water by
86.7%, fat, by 0.02%, and wet fiber by 1.66%. According to (Roni, Susanto, Pratama, &
Herawati, 2020), the presence of carbohydrate content means that pineapple skin can be
used as a base material for bioethanol production, so that coconut water and pineapple
skin have the potential to make bioethanol.
RESEARCH METHODS
The main ingredients are old coconut water and honey pineapple skin, the supporting
materials are distilled water, HCl, Urea NPK, Saccharomyces cerevisiae yeast, Glucose
Eduvest Journal of Universal Studies
Volume 1 Number 6, June 2021
425 http://eduvest.greenvest.co.id
Anhydrous, Anthrone Sulfate. In this study, using the method of fermentation with the
beginning of the process of pre-treatment of raw materials, the analysis of glucose,
fermentation and distillation.Method used factorial design, the control variables used are
sample volume 5 ml, Anthrone 20 mg, 1 mg anhydrous glucose, wavelength uv-vis
spectrophotometer 630nm, sample volume 150 ml coconut milk, pineapple filtrate
volume of 150 ml, distillation temperature 78
0
C - 80
0
C. The independent variables used
were fermentation time of 24 hours and 72 hours, temperature of 26
0
C and 30
0
C, yeast
concentration of 5 grams and 2 grams, urea NPK 3 grams.
Table 1 The experimental design with a factorial design
Description of table 1:
Time (t): (-) 24 and (+) 72 hours
pH: (-) 4 and (+) 5
Yeast content (k): (-) 5 and (+) 2 grams
The research process can be observed in Figure 1 analyzes were performed on the
samples to determine the glucose level analysis with Quicker-Method method, and
product analysis is performed to determine the density, calorific value, ethanol content,
and Specific Gravity.
No.
T
tpH
Tk
pHk
tpHk
1.
-
+
+
+
-
2.
+
-
-
+
+
3.
-
-
+
-
+
4.
+
+
-
-
-
5.
-
+
-
-
+
6.
+
-
+
-
-
7.
-
-
-
+
-
8.
+
+
+
+
+
Rizka Lestari Dewi and Anggun Puspitarini Siswanto
Utilization of bioetanol fermentation waste pineapple and coconut water as
disinfectants with bacteria saccharomyces cerevisiae 424
Figure 1. Flowchart of Research
Rizka Lestari Dewi and Anggun Puspitarini Siswanto
Utilization of bioetanol fermentation waste pineapple and coconut water as
disinfectants with bacteria saccharomyces cerevisiae 426
The Athrone method was used to determine the sugar content in honey and old
coconut water samples, and the analysis of variants using the Quicker Method.
RESULTS AND DISCUSSION
A. Analysis of Glucose Levels Using the Anthrone Method
The standard solution is made of 6 concentrations, namely 0 mg / ml; 0.2 mg / ml;
0.4 mg / ml; 0.6 mg / ml; 0.8 mg / ml; 1 mg / ml made from the dilution of 0.2 mg
glucose standard solution and then take 10 ml dissolved in 100 mL volumetric flask. The
following chart calibration curve.
Figure 2. Calibration Curve Graph
From the graph, obtain standard solutions absorbance measurement results are
then used to create a standard curve. Standard curves need to be created to determine the
regression equation. So that the regression equation can be determined the sample
concentration. The regression equation obtained is y = 0.08x + 0.13 with R
2
= 0.905.
Based on the regression equation that has been made, it can be determined the
glucose level in honey pineapple skin, old coconut water and honey pineapple skin + old
coconut water. The glucose level in the sample of honey pineapple peel as a result of the
experiment was 8.24 mg / mL, the sample of old coconut water was 8.23 mg / mL, and in
the sample of pineapple honey + old coconut water was 8.11 mg / mL.
While the glucose content in honey pineapple peel is 8.53%, the glucose content in
old coconut water is 3%. In this study, the percentage of glucose levels for honey
pineapple peel was 0.08% and in old coconut water was 0.08%. The small glucose level
in the experimental sample is likely due to the influence of human error and the error in
the absorbance measurement of the standard solution as discussed earlier, so that the
standard curve equation is not good and affects the determination of glucose levels in the
sample.
y = 0.0878x + 0.1316
R² = 0.9051
0
0.2
0.4
0.6
0.8
1 2 3 4 5 6
Konsentrasi Gula
Standar(mg/ml)
Absorbansi (A)
Data Kalibrasi
Linear (Data Kalibrasi)
Rizka Lestari Dewi and Anggun Puspitarini Siswanto
Utilization of bioetanol fermentation waste pineapple and coconut water as
disinfectants with bacteria saccharomyces cerevisiae 428
B. Analysis of Variants Using the Quicker Method
a. Effect of Density Variables on Bioethanol
Table 2 Calculation Results Main Effects and Interactions Effects of Ethanol Density
Table 2 shows that the main effect for the density value in this study is the
fermentation time (t) with a value of 0.01 with an interaction effect in the form of pH and
nutrient mass with a value of -0.04.
Table 3. Determination of Variables Influencing Bioethanol Density
P (%)
Effects
Identity Effects
7,14
0,04
M
21,42
0,01
P
35,71
0.01
T
50
0,02
tP
64,29
-0,03
Tm
78,58
0,01
tPm
92,86
-0,03
Pm
Figure 3. Normal Probability Density Plot Against Bioethanol for Factorial Design 2
3
Figure 3 shows the Normal Probability Plot graph between the P value and the
effect obtained by regression (R
2
) of 0.85 by activating the Trendline feature in Microsoft
Excel. This means that 85.63% of the total variation in the model can be represented by a
regression equation. The equation that shows the correlation between the value of
bioethanol density and the parameters of the research process (nutrient mass and
Effects
Results
t
0,01 Main Effects
P
0,01
m
0,04
tP
0,02
tm
-0,03
Pm
-0,03 Interactions Effects
tPm
0,01
y = 973.02x + 46.108
R² = 0.8563
0
20
40
60
80
100
-0.06 -0.04 -0.02 0 0.02 0.04 0.06
P(%)
Effects
Eduvest Journal of Universal Studies
Volume 1 Number 6, June 2021
427 http://eduvest.greenvest.co.id
operating conditions) is y = 973.02x + 46.10.
b. Effect of Variable Ethanol Levels on Bioethanol
Table 4. Calculation Results of Main Effects and Interaction Effects on Ethanol
Bioethanol Levels
Table 4 shows that the main effect for the value of ethanol content in this study is
the fermentation time (t) with a value of -0.05 with an interaction effect in the form of pH
and nutrient mass with a value of 0.23.
Table 5 Determination of Influential Variables on Bioethanol Ethanol Levels
P (%)
Effects
Identity Effects
7,14
-0,27
M
21,42
-0.09
P
35,71
-0.05
T
50
-0,15
tP
64,29
0,23
Tm
78,58
-0,07
tPm
92,86
0,23
Pm
Figure 4. Normal Probability Plot Graph of Ethanol Bioethanol Content for Factorial
Design 2
Figure 4 shows the Normal Probability Plot graph between the P value and the
effect obtained by regression (R2) of 0.89 by activating the Trendline feature in Microsoft
Excel. This means that 89.16% of the total variation in the model can be represented by a
regression equation. The equation that shows the correlation between bioethanol ethanol
Effects
Results
t
-0,05 Main Effects
P
-0,09
m
-0,27
tP
-0,15
tm
0,23
Pm
0,23 Interaction Effects
tPm
-0,07
y = 155.71x + 53.746
R² = 0.8916
0
20
40
60
80
100
-0.30 -0.20 -0.10 0.00 0.10 0.20 0.30
P(%)
Effects
Eduvest Journal of Universal Studies
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429 http://eduvest.greenvest.co.id
content and the parameters of the research process (nutrient mass and operating
conditions) is y = 155.71x + 53.74.
c. Effect of Variable Specific Gravity (sg) on Bioethanol
Table 6 Calculation Results of Main Effects and Interaction Effects on Specific Gravity
(sg) Bioethanol
Effects
Results
t
0,01 Main Effects
P
0,02
m
0,04
tP
0,02
tm
-0,04
Pm
-0,04 Interaction Effects
tPm
0,01
Table 6 shows that the main effect for the specific gravity (sg) value in this study
is the fermentation time (t) with a value of 0.01 with an interaction effect in the form of
pH and nutrient mass with a value of 0.01.
Table 7 Determination of Variables Affecting the Specific Gravity (sg) of Bioethanol
P (%)
Effects
Identity Effects
7,14
0,04
M
21,42
0,02
P
35,71
0,01
T
50
0,02
tP
64,29
-0,04
Tm
78,58
0,01
tPm
92,86
-0.04
Pm
Figure 5. Normal Probability Plot Against Specific Gravity (sg) Bioethanol for Factorial
Design 2
3
Figure 5 shows the Normal Probability Plot graph between the P value and the
effect obtained by regression (R2) of 0.87 by activating the Trendline feature in Microsoft
Excel. This means that 86.6% of the total model variation can be represented by a
regression equation. The equation showing the correlation between the specific gravity
y = 975.42x + 46.086
R² = 0.866
0
10
20
30
40
50
60
70
80
90
100
-0.060 -0.040 -0.020 0.000 0.020 0.040 0.060
P(%)
Effects
Eduvest Journal of Universal Studies
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431 http://eduvest.greenvest.co.id
(sg) of bioethanol and the parameters of the research process (nutrient mass and operating
conditions) is y = 957.42x + 46.08.
d. Effect of Variable Calorific Value on Bioethanol
Table 8 Calculation Results of Main Effects and Interaction Effects on Calorific Value of
Bioethanol
Table 8 shows that the main effect for the calorific value of bioethanol in this
study is the fermentation time (t) with a value of 32.07 with an interaction effect in the
form of pH and nutrient mass with a value of 169.16.
Table 9 Determination of Variables Affecting the Calorific Value of Bioethanol
P (%)
Effects
Identity Effects
7,14
-190,14
M
21,42
-109,33
P
35,71
32,07
T
50
-133,69
tP
64,29
54,86
Tm
78,58
33,24
tPm
92,86
169,16
Pm
Figure 6. Normal Probability Plot Graph of the Calorific Value of Bioethanol for
Factorial Design 2
3
Figure 6 shows the Normal Probability Plot graph between the P value and the
effect obtained by regression (R2) of 0.94 by activating the Trendline feature in Microsoft
Excel. This means that 94.18% of the total model variation can be represented by a
regression equation. The equation that shows the correlation between the calorific value
Effects
Results
t
32,07 Main Effects
P
-109,33
m
-190,14
tP
-133,69
tm
54,86
Pm
169,16 Interaction Effects
tPm
-33,24
y = 0.2358x + 54.845
R² = 0.9418
0
20
40
60
80
100
-300.00 -200.00 -100.00 0.00 100.00 200.00
P(%)
Effects
Rizka Lestari Dewi and Anggun Puspitarini Siswanto
Utilization of bioetanol fermentation waste pineapple and coconut water as
disinfectants with bacteria saccharomyces cerevisiae 430
of bioethanol and the parameters of the research process (nutrient mass and operating
conditions) is y = 0.23x + 54.84.
C. Relationship between Yeast Mass and Time on Bioethanol Density
Figure 7. Graph of Yeast Mass Relation to Bioethanol Density
Figure 8. Graph of the Relationship between Operating Conditions and Density of
Bioethanol
Figure 7, it can be seen that the density of bioethanol obtained is 0.96 gr / ml, where
the density exceeds the requirements for the quality requirements of bioethanol, namely
0.82 gr / ml. This shows that the ethanol produced is still not pure because ethanol is
mixed with water. This is because the distillation does not maintain the stability of the
distillation temperature, so that the ethanol that comes out has been mixed with water. In
Figure 8, it can be seen that 24 hours and 72 hours of fermentation using Saccharomyces
Cerevisiae yeast. This shows that 24-hour ethanol with a pH of 4 contains more ethanol
than 72 hours with a pH of 5. This is due to the lack of density in the fermenter bottle, the
theory is that the longer the fermentation time, the more ethanol content is due to the
activity of
0.925
0.93
0.935
0.94
0.945
0.95
0.955
0.96
0.965
0.97
0.975
1 dan 5 2 dan 6 3 dan 7 4 dan 8
DENSISTAS (gr/ml)
RUN
5 gr/l
2 gr/l
0.92
0.94
0.96
0.98
5 2
DENSITAS (gr/ml)
MASSA RAGI (g)
t = 24 jam dan pH = 4 t = 72 jam dan pH = 4
t = 24 jam dan pH = 5 t = 72 jam dan pH = 5
Eduvest Journal of Universal Studies
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Rizka Lestari Dewi and Anggun Puspitarini Siswanto
Utilization of bioetanol fermentation waste pineapple and coconut water as
disinfectants with bacteria saccharomyces cerevisiae 432
Saccharomyces Cerevisiae experiencing a phase stationary, where there is a process of
breaking down glucose on a large scale. The results of the breakdown of glucose by
Saccharomyces Cerevisiae produce ethanol.
D. Relationship between Yeast Mass and Fermentation Time on Bioethanol Levels
Figure 9. Graph of Yeast Mass Relationship to Bioethanol Ethanol Content
Figure 10 Graph of the Relationship between Operating Conditions and Bioethanol
Ethanol Content
Figure 9 and Figure 10, it can be seen that the highest bioethanol content is 32%
with a yeast mass of 5 grams for 24 hours. However, at 72 hours with a mass of 5 g of
yeast it has a bioethanol content of 27%. This shows that the bioethanol content is not in
accordance with the provisions of the bioethanol quality requirements, namely 94.1%
minimum due to the lack of density in the fermenter bottle and less maintaining
temperature stability in distillation. This is not in accordance with the theory, the real
theory is that the longer the fermentation time, the higher the bioethanol content.
E. Relationship between Yeast Mass and Fermentation Time on Bioethanol
Calorific Value
0%
10%
20%
30%
40%
1 dan 5 2 dan 6 3 dan 7 4 dan 8
KADAR ETANOL (%)
RUN
5 gr/l
2 gr/l
0%
20%
40%
5 2
KADAR ETANOL (%)
MASSA RAGI (g)
t = 24 jam dan pH = 4 t = 72 jam dan pH = 4
t = 24 jam dan pH = 5 t = 72 jam dan pH = 5
0
50
100
150
200
250
1 dan 5 2 dan 6 3 dan 7 4 dan 8
NILAI KALOR (Kkal/kg)
RUN
5 gr/l
2 gr/l
Eduvest Journal of Universal Studies
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433 http://eduvest.greenvest.co.id
Figure 11 Graph of Yeast Mass Relationship to Bioethanol Calorific Value
Figure 12. Graph of the Relationship of Operating Conditions to the Calorific Value of
Bioethanol
In Figure 11 and Figure 12, the highest heating value at 72 hours is 211.95 kcal / kg.
The calorific value obtained in bioethanol produced from pineapple skin honey and old
coconut water is still small compared to bioethanol from different raw materials,
including the calorific value of organic waste ranging from 10,000-11,000 kcal / kg. A
higher calorific value will make it more flammable so that the quality of bioethanol is
more flammable. This shows that the quality of bioethanol produced in this study is still
low, this is due to the lack of density in the fermenter bottle and the lack of temperature
stability in the distillation.
F. Relationship between Yeast Mass and Fermentation Time Against Specific
Gravity Bioethanol
Figure 13 Graph of Yeast Mass Relationship to Bioethanol Specific Gravity
0
100
200
300
5 2
NILAI KALOR (Kkal/kg)
MASSA RAGI (g)
t = 24 jam dan pH = 4 t = 72 jam dan pH = 4
t = 24 jam dan pH = 5 t = 72 jam dan pH = 5
0.92000
0.94000
0.96000
0.98000
1 dan 5 2 dan 6 3 dan 7 4 dan 8
SPECIFIC GRAVITY (SG)
RUN
5 gr/l
2 gr/l
0.90000
0.95000
1.00000
5 2
SPECIFIC GRAVITY (SG)
MASSA RAGI (g)
t = 24 jam dan pH = 4 t = 72 jam dan pH = 4
Rizka Lestari Dewi and Anggun Puspitarini Siswanto
Utilization of bioetanol fermentation waste pineapple and coconut water as
disinfectants with bacteria saccharomyces cerevisiae 434
Figure 14 Graph of the Relationship of Operation Conditions to the Specific Gravity of
Bioethanol
In Figure 13 and Figure 14 the results of the highest specific gravity at 24 hours
and the mass of 4 gr yeast are 0.98 gr / ml, where the specific gravity exceeds the
requirements for the quality requirements of bioethanol, namely 0.82 gr / ml. From the
research conducted, the length of fermentation has an influence on the specific gravity of
the alcohol being tested, this effect is in the form of a decrease in the specific gravity
value with increasing time. The theory should be that the longer the fermentation lasts,
the number of microbes needed in the process will also increase, so that with the
increasing number of microbes, the more carbohydrates are broken down into alcohol.
With the increase in the amount of alcohol, automatically the weight or density of
the alcohol-water mixture will be lower, which also causes the specific gravity of the
mixture to have a low value. In addition, temperature also has an influence on specific
gravity, the relationship between specific gravity and temperature, the higher the
temperature, the higher the value of specific gravity. This is because when the distillation
is carried out at a high temperature, the amount of water that accompanies the alcohol
will also be high and vice versa and therefore the density of the distillation product will
also be high.
G. Comparison of Bioethanol Quality Standards
Table 10 Quality Requirements for Bioethanol
Parameter
Unit
Bioethanol
Standard
Quality
Pineapple
Skin +
Coconut
Water
Bioethanol
Information
Ethanol
Content
% v/v
Min 94,1
32
Not appropriate
Fusel Oil
mg/L
Max 15
-
-
Aldehid
mg/L
Max 30
-
-
Metanol
mg/L
Max 30
-
-
Density
gr/ml
Max 8,21
0,97
Not appropriate
Specific
Gravity
-
Max 8,21
0,98
Not appropriate
Calorific Value
kkal/kg
Max 5000
211,95
Not appropriate
Acidity (as
acetic acid)
mg/L
Max 30
-
-
Water Content
% b/b
Max 2
-
-
Source: Badan Standar Nasional, Etanol Nabati, SNI 3565
Based on the bioethanol quality requirements, the bioethanol produced in this
study is not in accordance with the predetermined bioethanol quality requirements, this is
due to the absence of nutrient provision to decomposing bacteria so that it does not work
optimally in converting glucose into bioethanol. In addition, the accumulation of products
can accelerate the death of bacteria during fermentation. In this study, the lack of
attention was paid to bioethanol for the purity of the yeast used and for the suboptimal
purification process.
Eduvest Journal of Universal Studies
Volume 1 Number 6, June 2021
435 http://eduvest.greenvest.co.id
CONCLUSION
The results of this study are the results of the glucose content in honey pineapple
peel is 0.08%, the glucose content in old coconut water is 0.08%, but based on the theory,
the glucose content of honey pineapple peel is 8.53% and in old coconut water. 3%. So
the results of glucose levels are not in accordance with this theory. The yield of
bioethanol density was 0.96 gr / ml, but the density exceeded the requirements for the
quality of bioethanol, namely 0.82 gr / ml. The highest yield of bioethanol content is 32%
with a yeast mass of 5 grams for 24 hours. However, at 72 hours with a mass of 5 g of
yeast it has a bioethanol content of 27%.
The result of the highest heating value at 72 hours is 211.95 kcal / kg. The result
of the highest specific gravity at 24 hours and the mass of 4 gr yeast is 0.98 gr / ml, where
the specific gravity exceeds the requirements for the quality of bioethanol, namely 0.82 gr
/ ml. Based on these bioethanol quality requirements, the bioethanol produced in this
study is not yet in accordance with the predetermined bioethanol quality requirements,
this is due to the absence of nutrient provision to decomposing bacteria so that it is not
optimal in converting glucose into bioethanol.
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