THE USAGE OF SOURSOP (Annona muricata Linn.

) AS A
MEDIA IN MAKING NATA

INVENTION PROJECT REPORT

Written by
Eki Desi Handayani
XII SCIENCE C

SMA SUGAR GROUP

SITE PT GULA PUTIH MATARAM
LAMPUNG TENGAH
2013
 AUTHENTICATION SHEET

Title : The Usage of Soursop (Annona muricata Linn.) as a Media in

Making Nata

Written by : Eki Desi Handayani

Class : XII Science C

It has checked and agreed by supervisor and examiners on 07 November 2013.

Supervisor 1st Examiner 2nd Examiner

Henry T. Saragi Joko M. Nur M. Firmansyah

Invention Teacher

Krisanti

1
 PREFACE

The writer would like to acknowledge her countless thanks to the Most
Gracious and the Most Merciful, Allah SWT who always gives her all the best of this
life and there is no doubt about it. Shalawat and Salaam to the Prophet Muhammad
SAW and his family. This invention project report is presented to fulfill one of the
requirements in accomplishing writers studying at Sugar Group Senior High School.
The writer would like to take the opportunity to express her deep and sincere
gratitude to the following:
1. Beloved father and mother for the biggest prayer, support and motivation.
2. Mr. Henry T. Saragi is as a supervisor who has supervised, and guided the
writer patiently in writing the invention project journal, and also who has
given the writer instruction, then the knowledge and meaningful suggestion
for the writer.
3. Mrs. Krisanti is as an invention teacher who has given suggestion for the
correction of writing this invention project report.
4. Companions of 2013, all of civitas academica in SMA Sugar Group and to
anyone that can not be mentioned directly or indirectly who helped the writer
in completing this invention project journal.
The writing of this invention project report is an effort to optimize the writer as
best facilities in developing knowledge. The writer does appreciate any opinion, and
suggestion for the improvement of this invention project report.
And last, the writer hope that the invention project report can be as an
inspiration to find a great idea in the next.

Lampung, 24 October 2013

Writer

TABLE OF CONTENT

Page
VALIDATION SHEET i
PREFACE ii

2
TABLE OF CONTENT iii
LIST OF PICTURE iv
LIST OF TABLE vi
ABSTRACT vii
CHAPTER I INTRODUCTION
1.1 Background 1
1.2 Problem Formulation 1
1.3 Objective 1
1.4 Benefit 1
1.5 Hypothesis 1
CHAPTER II LITERATURE REVIEW
2.1 Soursop 3
2.2 Acetobacter xylinum 4
2.3 Nata 8
2.4 Fermentation 10
CHAPTER III METODHOLOGY
3.1 Time and Place 12
3.1 Tools and Materials 12
3.3 Procedure 13
CHAPTER IV DATA AND ANALYSIS
4.1 Data 20
4.2 Analysis 21
CHAPTER V CONCLUSION AND SUGGESTION
5.1 Conclusion 32
5.2 Suggestion 32
BIBLIOGRAPHY

LIST OF PICTURE

Page
Picture 1 Soursop (Anonna muricata Linn.) 1
Picture 2 Acetobacter xylinum 5
Picture 3 Layer of Nata de Coco 8
Picture 4 Separating Seeds from Flesh Soursop 13
Picture 5 Soursop Blended 13
Picture 6 Pouring Water 13
Picture 7 Filtering 14

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Picture 8 Soaking Tray in Boiled Water 14
Picture 9 Heating Tray above Fire 14
Picture 10 Heating newspaper above Fire 15
Picture 12 Rubber 15
Picture 13 Bean Sprout Ready to be Boiled 15
Picture 14 Filtering Boiled Bean Sprout 16
Picture 15 Bean Sprout Extract 16
Picture 16 Pouring Soursop Juice into Stewpan 17
Picture 17 Pouring Bean Sprout Extract 17
Picture 18 Adding Sugar 17
Picture 19 Pouring Acetic Acid 17
Picture 20 Adding Yeast 17
Picture 21 Boiling Material Mixture 18
Picture 22 Pouring Media into Tray 18
Picture 23 Cooling the Temperature of Media 18
Picture 24 Inoculation (Giving Nata De Coco Starter) 19
Picture 25 Fermentation 19
Picture 26 Filtering the Mixture of Soursop Water 22
Picture 27 Filtering Result (Soursop Juice) 22
Picture 28 The Second Experiment (zoomed in) 25
Picture 29 The Second Experiment Result (top view) 25
Picture 30 The Fourth Experiment Result (top view) 26
Picture 31 The Fourth Experiment Result (upright, front view) 26
Picture 32 Yeast 27
Picture 33 The Fifth Experiment Result (top view) 28
Picture 34 The Fifth Experiment Result (Nata Layer) 28
Picture 35 The Fifth Experiment Result (layer is raised) 29
Picture 36 Beverage of Soursop Mixture Sunglap Juice 29
Picture 37 The Sixth Experiment Result 30
Picture 38 Cooling Media to the Room Temperature 31

4
 LIST OF TABLE

Page
Table 1 Nutrient Content of Soursop /100 grams 3
Table 2 Time of Doing the Experiment 12
Table 3 The Result of 8x Experiments 21

5
 ABSTRACT

The Usage of Soursop (Anonna muricata Linn.) as a Media in Making Nata

By: Eki Desi Handayani

Soursop is a high nutritious fruit that have a lot of special quality and benefits.
The most nutrient content in soursop is carbohydrate. Whereas special quality and
benefit of soursop inter-alia are for medication, as a base material of food and
beverage industry, and it is as a produk olahan masyarakat. However, soursop is a

6
perishable fruit and it cannot be kept at longer time so as it needed anoter alternative
in the utilization. The alternative is to make Nata from soursop (Anonna muricata
Linn).
In this research, writer do the experiment by utilizing soursop juice in making
Nata, then it did the fermentation by adding Nata De Coco starter that is Acetobacter
xylinum. Writer did the several times of experiment and repeat to get the result in
making nata from soursop.
This research was did since on February until October 2013. Writer did the five
times of experiment, however it is only on the fifth experiment is success. Based on
the fifth experiment, it known that Nata can be made from soursop by blending 600
mL of soursop juice, 125 mL of bean sprout extract, 50 mL of Nata starter, 12,5 mL of
acetic acid, 75 g of sugar, and 0,125 g of yeast.

Keyword: soursop, Nata, Acetobacter xylinum

7
 CHAPTER I INTRODUCTION

1.1 Background
Soursop is a high nutritious fruit that have a lot of efficacies and benefits.
The highest nutrient content in soursop is a carbohydrate. One type of
carbohydrate in soursop is a reducing sugar, there are glucose and fructose with
levels of 81,9% 93,6% of the total sugar content (dr. Abu Hana). While
efficacies and benefits in soursop are for treatment, as the base material of food
and beverage industri, and it is as the society processed product. However,
soursop is a perishable fruit and it cannot be store in a long span of time so as it
needed another alternative in its utilization. The alternative is to use soursop as a
media in making Nata.

1.2 Problem Formulation

Does soursop can be used as a media in making Nata?

1.3 Objective
Try to use soursop as a media in making Nata.

1.4 Benefit
Give the information to society about the usage of soursop, it is not only to be
consumed commonly but it can be used as a material in making Nata.

1.5 Hypothesis
Soursop can be used as a media in making Nata because soursop content of
simple carbohydrate compound those are glucose and fructose.

1
 BAB II. LIST OF LITERATURE

2.1 Soursop
According to Sugeng Haryanto who is an author book of Encyclopedia of
Medical Plants said that soursop (Anonna muricata Linn.) is a plant or tree
which have small and low steam. The leaves are oval and rather thick, the top
surface is dark green colored, while the down surface has a younger color.
Soursop name itself comes from the Dutch language "Zuurzak" which roughly
means the acid sac (2009, 485:486)

Picture 1. Soursop (Annona muricata Linn.)

(Source: Maria, 2013)
In plant systematic (taxonomy), soursop is classified as follows:
Kingdom : Plantae
Divisi : Spermatophyta
Sub Divisi : Angiospermae
Class : Dicotyledonae
Ordo : Polycarpiceae
Famili : Annonaceae
Genus : Annona
Species : Annona muricata Linn.

As Drs. Hendro Sunarjono said (author of Crop Production Science and

Fruits) that soursop are consists of 67% fruit flesh that can be consumed, 20% of
peel, 8.5% of seed, and 4.5% of central axis of the fruit. Soursop has a fairly high
sugar content that is 68% of all solid parts of the fruit flesh. Soursop flesh is
white, soft, and has a lot of fibers and vitamins. Nutrient content contained in
soursop can be seen in the table below:

Table 1. Nutrient Content of Soursop/100 grams

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 (Source: Nutrition directorat and food of Health Department)

Composition Content
Energy (cal) 65
Protein (g) 1
Fat (g) 0,3
Carbohydrate (g) 16,3
Calcium (mg) 14
Phosphorus (mg) 27
Ferum (mg) 0,6
Fiber (g) 2
Vitamin A (SI) 10
Vitamin B (mg) 0,07
Vitamin C (mg) 20
Niasin (mg) 0,7
Water (%) 81,7

Based on the article entitled Content, Efficacy, and Benefit of Leave and
Soursop by dr. Abu Hana, the highest content of nutrients in the soursop is
carbohydrate. One type of carbohydrate in soursop is a reducing sugar those
are glucose and fructose levels of 81.9% to 93.6% of the total sugar content.
Hasan Aroni (one of the workers in Malang Healthy Polytechnic) is
adding that soursop contains a very little fat (0.3 g/100 g), so it is good for
healthy. Sour taste in soursop derived from non-volatile organic acids,
particularly malic acid, citric acid, and acid isositrat.
The most dominant vitamin in soursop is vitamin C, which is about 20
mg/100 g of fruit flesh. Vitamin C requirement of every person per day (60 mg),
have been met by only consuming 300 g of soursop flesh. The content of
vitamin C which is high enough on soursop is an excellent antioxidant to
increase endurance and decelerate the aging process (stay young).

The fairly dominant of mineral are phosphorus and calcium, respectively

27 mg and 14 mg/100 g. Both minerals are essential for the formation of bone
mass, so it is useful to form strong bones and prevent osteoporosis. Beside
nutrient component, soursop is also very rich in non-nutritional components.
One of them is containing lots of fiber food (dietary fiber), which reached 3.3
g/100 g of fruit flesh.

3
 dr. Abu Hana has said that one type of carbohydrate in soursop is a
reducing sugar (glucose and fructose) with levels of 81.9 to 93.6 percent of the
total sugar content. Fructose is a simple sugar (monosaccharide) of plants,
especially those contained in the fruit, a sugar with atom C (carbon) 6
(heksosa), slightly sweeter than cane sugar (sucrose), and it is important for
diabetes mellitus sufferer because it will not increasing the blood sugar-content
when eaten (Wildan Yatim, 1999, 376)
The richest nutrient content in soursop can make its fruit is used a lot of
processed food products. As Gabrielle Nindya said (a student university of the
Medical Faculty of Nutrition Program at the Diponegoro University). In a paper
entitled Soursop and Vegetable, there are the result of processed products such
as soursop juice, dodol, jam, and candied soursop (2011, 7:9). Despitefully,
according to Elwin Resie (worker of BBPP (Center for Agricultural Training
Malang) is giving additional information about other processed products from
soursop named candy and fruit leather.
There are some characteristics of ripe and rotten soursop. Ripe soursop
is shown by soft, spines on the fruit does not feel sharp, yellowed stalk, and
good smelled. While rotten soursop can be shown by peel is black-brown, foul-
smelling and slimy, and also it is flabby when touch.

2.2 Acetobacter xylinum

Picture 2. Acetobacter xylinum

(Source: Muh. Taufiq Munawar, 2009)

Taxonomy of Acetobacter xylinum:

Kingdom : Bacteria
Phylum : Proteobacteria
Class : Alpha Proteobacteria

4
Ordo : Rhodospirilia
Famili : Pseudomonadaceae
Genus : Acetobacter
Spesies : Acetobacter xylinum
(Adha Panca Wardanu, 2011)
Based on the working paper written by a student of North Sumatera
University, Acetobacter xylinum has 2 characteristics including morphology and
physiological characteristics.
1. Morphological Characteristic
Acetobacter xylinum is a short rod-shaped bacteria, which has a length
are 2 microns and 0.6 microns wide, with a slimy surface of the wall. The
bacteria can form short chains with 6 - 8 unit cells. These bacteria does not
form endospores or pigment. In the young cell culture, the individual cells are
independently and transparently. Old colonies form a solid cover resembles
gelatin and cell colonies. Colony growth in liquid medium after 48 hours of
inoculation will establish pellicle layer and can be easily retrieved with a
needle oasis.

2 .Physiological Characteristic
Acetobacter xylinum can form acid from glucose, ethyl alcohol, and
propel alcohol, it does not form indol, and it has the ability to oxidize acetic
acid into CO2 and H2O. The most prominent characteristic of these bacteria is
the ability to polymerize glucose into cellulose. Next, the cellulose matrix form
known as Nata. Dominant factors that influence to the physiological
characteristics of Nata formation are the availability of nutrients, acidity,
temperature, and oxygen availability.
There are some factors that can affect to the growth of Acetobacter
xylinum including the following:
a. Carbon source
Carbon source that can be used in the fermentation of Nata is a
carbohydrate compounds are classified as monosaccharide and
disaccharides. Nata formation can occur in media containing the compounds
of glucose, sucrose, and lactose. While, the most widely used is based on
economic considerations is sucrose or sugar. The addition of sucrose should
refer to the amount needed. The excessive addition can affect to the Nata
forming, it is uneconomic and also create a new form of residual waste from

5
 sucrose. But on the contrary, the much less addition can cause a Nata starter
grow abnormally and can not be produced maximally.
b. Nitrogen source
Nitrogen sources can be used from organic and inorganic compounds.
Good material for the growth and formation of Acetobacter xylinum and Nata
forming are yeast extract and casein. However, ammonium sulfate and
ammonium phosphate (in the market known as ZA) are a material which is
more suitable from the point of economics view and the quality of the
resulting Nata. There are many other N sources that can be used and
inexpensive as urea.
c. The level of acidity (pH)
Although it can be grown in a pH range from 3.5 to 7.5, Acetobacter
xylinum is suitable to grow in acidic conditions (pH 4.3). If the environmental
conditions under bases conditions, the bacteria will get disturbance in its cell
metabolism.

d. Temperature
The ideal temperature (optimum) for the growth of bacteria Acetobacter
xylinum is 28C - 31C. The temperature range is room temperature. At
temperatures below 28C, bacterial growth is inhibited. Similarly, at
temperatures above 31C, Nata will be damaged and even death, although
extracellular enzymes that have been produced to keep working to form
Nata.

e. Air (Oxygen)
Acetobacter xylinum is an aerobic bacteria. In its growth, propagation,
and activities, the bacteria need oxygen. These bacteria will be impaired in its
growth and will die immediately if it deprived of oxygen. Therefore, the
container used for fermenting Nata De Coco should not be sealed closely. To
fulfill the oxygen need, the place for Nata fermentation provided the
ventilation enough.
If Acetobacter xylinum grown in a media which is rich of sucrose (sugar),
these bacteria will break down the sucrose into glucose and fructose. The

6
 compounds of glucose and fructose are consumed as an ingredient for the new
cell metabolism.
Acetobacter xylinum remodel sugar to obtain energy necessary for cell
metabolism. Beside that, these bacteria also secrete enzymes capable to
preparing (polymerize) glucose into polysaccharide compounds known as
extracellular cellulose (Nata de Coco).
In other function, fructose is used as a source of energy, base materials
of Nata after hydrolyzed to become glucose, and also it functioned as inducers
for the synthesis of extracellular enzymes polymerase. This is one reason that
sucrose has advantage than other simple sugars in its function as a substrate
maker Nata.
Based on morphological observation, Nata formation by Acetobacter
xylinum bacteria begins with the formation of cellulose sheets threads.
Formation of the yarn at first is looks like a flagellum (whip bacteria).
Furthermore, the bacteria Acetobacter xylinum cellulose form mikrofibil around
his body to form the surface of the cellulose fibers are very numerous and
reaches a certain thickness. In the end, the cellulose structure will look like the
transparent white layer with the surface is slippery and smooth called Nata.
In the article entitled Nata De Coco posted by Gusti Stevani explained
that the process of formation of Nata De Coco by Acetobacter xylinum activity
can be described as follows : Acetobacter xylinum cells take up glucose from
the sugar solution, then combined with fatty acids to form precursors to the cell
membrane, then out together polymerizing enzyme glucose into cellulose
outside the cell. Precursors of these polysaccharides are GDP-glucose.
Precursor formation is stimulated by the presence of a catalyst such as Ca2+,
Mg2+. This precursor then undergoes polymerization and binds to the acceptor
forming cellulose or Nata.

2.3 Nata
According to the book entitled Scientific Biology Workbook for SMP 3B
written by Istamar Syamsuri, Nata is a collection of cellulose that has an
expansible texture, white, resembles a gel, and floating on the surface of the
liquid. Materials that can be used as a media in making Nata include coconut

7
water, so as the product is known as Nata De Coco. In addition, other materials
that can be used is the pineapple juice (Nata De Pina), soybean (Nata De
Soya), or other fruits that contain glucose. Bacteria is used in the manufacture
of Nata is Acetobacter xylinum. These bacteria can convert sugars into
cellulose. Cellulose is what makes the collection so it looks white Nata (2006,
65)

Picture 3. Layer of Nata de Coco

(Source: Rahmad Agus Koto, 2012)
Then, according to the working paper written by students at the North
Sumatra University, there are several stages in the manufacturing activities Nata
as follows:
1 . Preparation
Preparation stage consists of several activities as follows:
a) Filtering
Filtering aims to separate dirt or foreign objects mixed with coconut water,
such as the fiber rest. Better filtering is done when using a cloth filter.
Coconut water contains a lot of dirt and dust will produce Nata cloudy with
a less attractive appearance and seemed dirty.

b) The addition of sugar and ammonium sulfate

Availability of carbohydrates and proteins contained in coconut water
insufficient for the formation of Nata, the coconut water into sugar needs to
be added at least 2.5 % and 0.5% ammonium sulfate. The ratio of carbon
and nitrogen in making Nata is the main key to the process does not
produce waste in the form of residual liquid fermentation ( level achieved

8
 zero residual substrate ), substrate ( nutrients in the media ) is used by
Nata bacteria. Types of carbon sources can include materials such as
glucose, lactose, and fructose. Similarly, the type of nitrogen source used
may be organic nitrogen such as proteins and yeast extract, and inorganic
nitrogen such as ammonium sulfate, ammonium phosphate, and urea.

c) Boiling
Boiling is done to boil and maintained for 5-10 minutes to ensure that
microbial contaminants are dead, and also complete dissolution of sugar
and ammonium sulfate were added.

d) The addition of acetic acid

The purpose of adding acetic acid is to decrease the pH of coconut water
reached a pH around 6.5 to 4.3. But, 4.3 is the optimum condition for the
growth of Acetobacter xylinum.

e) Cooling
The best cooling accomplished by allowing the fluid in the tray for one
night. It is well to check the presence or absence of contaminants that
grow on the liquid.

2. Inoculation, Fermentation, and Handling

a) Providing Seed (Inoculation)
Provision of seeds is done when the mixture of coconut water, ammonium
sulfate, and acetic acid has been really cold. When giving seeds is done at
liquid coconut water still hot or warm, it can make Nata bacteria die, so
that the fermentation process can not last long.
b) Fermentation or Ripening
Mixture of coconut water that has been given a seed, left for 7-8 days for a
process of fermentation and formed Nata De Coco.

Nata fermentation media composition consists of carbohydrate (sugar) as the

carbon source and urea as nitrogen sources. Because it needs to be added such
substances include :
a) Sugar as a carbon source

9
 Carbon source is an important factor in the process of fermentation. Bacteria to
produce Nata require a carbon source for metabolic processes. Glucose to
enter the cells and it is used for energy supply needed for breeding. The amount
of added sugars should be considered so as sufficient for metabolism and
pellicle formation Nata. Carbon requirements for the media generally provided
by glucose, starch, and lactose (Hidayat, 2006)

b) Urea as a nitrogen source

Beside sugars, nitrogen sources is an important factor as well. Nitrogen is
needed for cell growth and enzyme formation. Nitrogen deficiency causes cells
to grow poorly and inhibit the formation of enzymes necessary, so that the
fermentation process may have failed or imperfect (Hidayat, 2006)

2.4 Fermentation
" Fermentation = fermentation . Remodel an organic molecule into
intermediates with the help of fungi, most of the yeast (Saccharomyces) . But
then besides yeast fungi also others, such as Rhizopus, and bacteria. Food and
beverage fermentation is tape, oncom, tempeh, tofu, beer, sake, wine, whiskey,
brandy, root bear, and butter " (Wildan Yatim, 1999, 376)
In the book of Fermentation Technology written by Suprihatin, fermentation
word comes from the Latin language "fervere" which means to boil. From the
Latin language meaning can be attributed to the condition of bubbly or boiling
liquids. This situation is due to the activity of the yeast on the extraction of fruits
or grains. Bubbles of carbon dioxide produced from the anaerobic catabolism of
sugars.
Definition of fermentation abstracted by Suprihatin that fermentation had
definition as s process of chemical changes in an organic substrate through the
activity of enzymes produced by microorganisms.

10
 CHAPTER III. METHODOLOGY

In this research, writer did an experiment by using soursop juice in making

nata, then fermented by adding Nata De Coco starter that is Acetobacter xylinum.
Writer did the experiment and repeated it in several times to get the results of Nata
of soursop .

1.1 Time and Place

1.1.1 Time
The time of the research held on February until October of 2013.
Table 2. Time of Doing the Experiment

Experimen
Date Time (WIB)
t
1 10-28 February 2013 16.30 19.00
2 30 Juni-13 July 2013 18.30 20.35
3 30 Juni-13 Juli 2013 18.30 20.35
4 24 August 06 September 2013 06.00 09.05
5 31 August - 21 September 2013 18.00 20.13
6 11 September 27 September 18.30 19.15
7 07 October 22 October 2013 15.00 17.30

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 8 22 October 05 November 2013 20.15 22.06
1.1.2 Place
The place of the research took place at PT SIL Housing 1 Block E 14.

1.2 Tools and Equipments

1.2.1 Tools: 3.2.2 Equipments
1. Stewpan 1. Ripe soursop (800 g)
2. Spatula 2. Sugar (75 g)
3. Scale 3. Bean sprout extract (125 mL)
4. Measuring glass 4. Acetic acid (12,5 mL)
5. Tray 5. Nata starter (50 mL)
6. Newspaper 6. Water (150 mL)
7. Yeast (0,125 g)
7. Blender
8. Dust-cloth
9. Gas stove
10.Rubber

3.3 Procedure
1. Preparation
1. Separate the seeds from flesh soursop, then take the flesh fruit only. The
weight of flesh fruit after measuring is 800 g.

Picture 4. Separating Seeds from Flesh Soursop

2. Pour 150 mL of water into flesh soursop, then blend it by using blender.

12
 Picture 5. Soursop Blended Picture 6. Pouring Water
3. Filter the mixture between soursop and water by using dust-cloth. After
that, measure the weight by using scale. The weight of soursop juice is
600 mL.

Picture 7. Filtering
4. Prepare a tray in dry condition and do the sterilization by soaking in boiled
water that has poured into receptacle, then heat it with distance is around
15 - 20 cm above fire, do it until 2 - 3 times of repetition. After that, put it
in face downward condition.

Picture 8. Soaking Tray in Boiled Water

13
 Picture 9. Heating Tray above Fire
5. Prepare some newspaper and do the sterilization by doing the same way
as step at number 4.

Picture 10. Heating Newspaper above Fire

6. Prepare some rubber to bind newspaper that will be used to close the tray
when fermentation process.

Picture 12. Rubber

2. Making Bean Sprout Extract

1. Boil 125 mL of water while adding 50 g of bean sprout by using stewpan.

14
 Picture 13. Bean Sprout Ready to be Boiled
2. Let it until 3 minutes, then smash it by using spoon.
3. Filter the boiled bean sprout by using dust-cloth so as separated from its
bean sprout. After the filtering, take the water only until got 125 mL of
bean sprout extract.

Picture 14. Filtering Boiled Bean Sprout

4. Use the bean sprout extract only.

Picture 15. Bean Sprout Extract

3. Using Soursop as a Media in Making Nata

1. Pour 600 mL of soursop, 125 mL of bean sprout extract, 12.5 mL of acetic
acid, and add 70 g of sugar into stewpan.

15
 Picture 16. Pouring Soursop Juice into Stwepan

Picture 17. Pouring Bean Sprout Extract Picture 18. Adding Sugar

Picture 19. Pouring Acetic Acid Picture 2. Adding Yeast

16
2. Boil the materials mixture.

Picture 21. Boiling Materials Mixture

3. After it boil, pour media into tray that already sterile with the dimension of
tray is 26.8 cm x 22 cm x 3.9 cm, then wait until 30 minutes. Next, cover it
with newspaper and bind with rubber.

Picture 22. Pouring Media into Tray

17
 Picture 23. Cooling the Temperature of Media
4. When it is still warm, pour Nata starter and no need to be stir. Cover itwith
newspaper but do not too close.

Picture 24. Inoculation (Giving Nata De Coco Starter)

5. Save the media in cupboard during 21 days at room temperature, that is
28-30C.

Picture 25. Fermentation

6. After 21 days, open the cover, then observe it.

Information: pictures comes from writer document source.

18
 CHAPTER IV.DATA AND ANALYSIS

4.1 Data
Table 3. Data of The Result Experiments

Experiment
Materials Result
/Date
Failed.
1/ Solid on the top layer, while the down
Soursop, bean sprout extract,
10
Nata starter, sugar, acetic acid layer is liquid, brown colored at some
28/02/2013
layer
Soursop juice 650 mL Failed.
Bean sprout extract 20 mL Doesnt form layer, liquid, bad
2/
30/06 Nata starter 25mL smelled, many small gibbosity and
Sugar 40 g
13/07/2013 yellow spot on the top layer, yellow
Acetic acid 15 mL
colored, mucous
Soursop water 650 mL
3/ Bean sprout extract 20 mL Failed.
30/06 Nata starter 25 mL The layer is very solid, very bad
18/07/2013 Sugar 50 g smelled, dark brown colored
Acetic acid 20 mL
Soursop water 500 mL Failed.
ZA 0,2 g Formed layer, layer is like a gel but
4/
24/08 Sugar 0,5 g easy to dissolve in water, malleable,
Nata starter 20 mL
6/09/ 2013 milky colored, sour smelled enough,
Acetic acid 10 mL
smooth on the top layer
Soursop juice 600 mL Success.
Bean sprout extract 50mL Formed layer, the layer is like a gel,
5/ Sugar 75 g expansible, and smooth on the top
31/08 Nata starter 50 mL
Acetic acid 12,5 mL layer, milky colored, and the thickness
21/09/ 2013
is less than 1 cm. But some layer get
Yeast 0,125 g
contamination or broken

19
 Percobaan
Bahan Hasil
ke-/Tanggal
Soursop (Beverage
500 mL Failed.
6/ Product)
Doesnt form layer, liquid, black-
11/09 Bean sprout extract 100 mL
Nata starter 20 mL brown colored, there are some black
27/11/2013 Sugar 50 g spot like fungi, very bad smelled
Acetic acid 10 mL
Soursop juice 500 mL Failed.
Bean sprout extract 125 mL Doesnt from layer, the layer is very
Nata starter 20 mL solid and the shape is irregular, curly
7/ Sugar 75 g
22/10 Acetic acid 15 mL shaped and dark brown colored, but
18/07/2013 some layer is yellow colored and it is
Yeast 5g malleable, a little mucous, very bad
smelled
Soursop juice 500 mL
8/ Bean sprout extract 125 mL
22/10 Sugar 100 g Totally failed.
Nata starter 20 mL
05/10/ 2013 Acetic acid 20 mL
Yeast
4.2 Analysis
Third table is a table contained of needed materials data and results in
every experiment of using soursop as a media in making Nata. Based on data
from the fourth table, the results of 8 experiments can be explained as follow:
1. First Experiment
In the first experiment conducted on 10 February 2013, writer used
ingredients such as soursop juice, bean sprout extract, Nata starter, sugar,
and acetic acid. To get soursop juice, the first step writer did is blending the
flesh soursop and water. After that, filtered by using gauze-filter to be took the
sunglap juice only.

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Picture 26. Filtering the Mixture of Soursop Water Picture 27. Filtering Result (Soursop Juice)

Filtering is done because the soursop contain of high fiber so that when
mixed with water, media is still very strong, therefore filtering is done to
obtained the aqueous media. However, gauze-filter has small pores, so it is
only 650 mL obtained only from the number of soursop used. Beside that,
how to get bean extract sprout can be seen on section 3 at the workings of
making bean extract.
The materials used include soursop juice, bean sprouts extract, Nata
starter, sugar, and acetic acid. Function of using these materials as follows:
Soursop juice is used as a media for making Nata, bean sprouts extract
serves as a source of organic nitrogen bacteria that used for its as the
needed nutrients, sugar or sucrose serves as a carbon source necessary
bacteria in the process of metabolism, acetic acid is used in decreasing the
pH of the media, and Nata starter (Acetobacter xylinum bacteria) used in the
manufacturing process capable of polymerizing glucose Nata due to be
cellulose.
The first experiment is called as pre-experiment, because procedure of
the experiment has not proper yet totally. The writer did not do the measuring
of the material amount required, just use them as desired. Firstly, soursop
juice, bean sprouts extract, Nata starter, sugar, and acetic acid are mixed.
Soursop juice that has been mixed is used for making Nata. Secondly, the
media boiled until boiling. After that, the media allowed to stand for
approximately 15 minutes at room temperature, then added Nata starter. This
is done because if the starter directly included in the media is still hot the
bacteria will die. The ideal temperature (optimum) for the growth of

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 Acetobacter xylinum is 28C - 31C. The temperature range is room
temperature. At temperatures below 28C, bacteria growth is inhibited.
Similarly, at temperatures above 31C, Nata starter will be damaged and
even death, although extracellular enzymes that have been produced to keep
working to form Nata. The next step is the media fermented for 18 days, that
is from 10 to 28 February.
The result of the first experiment is failed. Soursop juice is used as a
media for making Nata only solidify the top layer, while the bottom layer is still
liquid. In addition, some brown coating, but partly white like Nata. Failure is
caused because some layers are not well controlled so contaminated by
other bacteria. This contamination is characterized by brown color in some
layers. Then, the failure of another factors is the lack of necessary nutrients
the bacteria in the process of making Nata. The writer did not know what the
material is less because the writer did not do the measuring of the material
amount used . Therefore, in subsequent experiments the writer do the
measuring and do the sterilization of the tools used.
2. Second and Third Experiment
Writer conducted two experiments at the same time which for the second
and third experiments were conducted on 30 June 2013. In doing the second
experiment, the writer has been measuring the amount of material used and
the sterilization of the tools used in the fermentation process such as tray,
and newspaper. How to sterilization has been written in section 1 at
preparation.
The materials needed are still the same, as already said in the first
experiment. The materials used include soursop juice, bean sprouts extract,
Nata starter, sugar, and acetic acid. Number materials used of both
experiments are same, but differentiated only of using acetic acid, sugar and
length of fermentation. Second experiment use 15 mL of acetic acid, 40 g of
sugar, and fermentation time is 13 days. While the third experiment used 20
mL of acetic acid, 50 g of sugar, and the length of fermentation was 18 days.

In the first experiment , 650 mL of soursop water, 20 mL of bean sprouts

extract, 25 mL of Nata starter, 15 mL of acetic acid, and 40 g of sugar are
mixed into stewpan, and boil until boiling. Soursop water that has been mixed
with a medium is to use for making Nata. After boiling the media, the media is

22
poured on a tray that has dimension is 26.8 cm x 22 cm x 3.9 cm, then the
media cooled by letting the media in the tray for one night. It is good for
checking the presence or absence of contaminant that grow on the liquid.
It is different from previous experiments, in these experiments the writer
try to not filtering substrate. Viscous media just left, this is done to determine
the effect of the fiber concentration in soursop. Then, giving Nata starter was
did one day after boiling. Media silenced within 1 day with the condition
covered by newspaper. After that, there was a phase and then inoculated
fermentation process . The second experiment was fermented for 13 days ,
while the third trial for 18 days.
Results of both experiments failed . The second experiment failed
because nata starter used was contaminated with other bacteria in the air .
The contamination is marked with the color changing nata starter . It is
related to the age of the bacteria . Another factor that failure is the lack of
availability of oxygen . Cover used to cover the trays are made of plastic
material and closed too tightly so that oxygen can not enter into the tray .
Nata starter bacteria Acetobacter xylinum is an aerobic microbes . In growth ,
development , and activities , these bacteria require oxygen in sufficient
quantities . When deprived of oxygen , these bacteria will be impaired in its
growth and will soon experience even death . Therefore , the container used
for fermenting nata , should not be sealed .
The third experiment also failed , as well as the source of
contaminated nata , the fermentation process is too long , causing the media
to be dark brown and cause a very unpleasant smell ( sour ) . Long
fermentation process causes the nutrients available in the medium reduced
the activity of the bacteria so that bacterial activity stalled and eventually
become damaged media.
For the next experiment, the writer try to reuse newspaper as the
cover and use a new Nata starter, and also decrease the duration of
fermentation time.

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 Picture 28. The Second Resul Experiment (Zoomed in)

Picture 29. The Second Result (top view)

3. Fourth Experiment
In the fourth experiment conducted on 24 August 2013, the writer used
materials such as 500 mL of soursop juice, 20 mL of Nata starter, 10 mL of
acetic acid, 50 g of sugar, and using 0.2 g of ZA. In this experiment, the writer
replace the using of bean sprouts extract with ZA as an inorganic nitrogen
source. All the materials are mixed in stewpan, then heated to boil up. After
that, the media poured into the tray with the same dimension 26.8 cm x 22 cm
x 3.9 cm to be settled for a few minutes by covering the tray with dust-cloth.
After that, Nata starter poured into the media. Furthermore, the media is
fermented for 13 days. The duration of fermentation in this experiment is
shorter than the previous experiment, that is in the third experiment. As
previous experiment, writer is not doing the fermentation process Given the
failure of the previous experiment, the writer did not do again the long time
fermentation process.
In the fourth experiment, the writer re-do the filtering. Soursop that has
been mixed with water then blended, it taken the sunglap juice only by

24
 filtering. Filtering is done by using a filter like nylon fabric. The pores on its
fabric are very small and texture of soursop that has been blended was very
soft so the writer really tried to get distillation of soursop juice hardly.
The result of the fourth experiment was still failed. The result was not as
expansible as Nata characteristics generally. Layer was soft and the color was
slightly browned. However, layer was almost like jelly solidifies, the sour smell
has been reduced, and the texture was smooth. Failure caused the nutrients
need for bacteria activities was less.

Picture 30. The Fourth Experiment Result (top view)

Picture 31. The Fourth Experiment Result (upright, front view)

For the next experiment, the writer use a tool such as fabric filter that has
pores larger than nylon cloth and increase the amount of material used.

4. Fifth Experiment
In the fifth experiment conducted on 31 August 2013, the writer used
materials such as 600 mL of soursop juice, 50 mL of bean sprout extract, 50
mL of Nata starter, 12.5 mL of acetic acid, 75 g of sugar, and adding 0.125 g of
yeast. All materials are mixed in stewpan then heated until boil up. After that,
media is cooled until reach to the room temperature that is 28C 31C.

25
Cooling media is done by letting it in stewpan, then stewpan is covered with
dust-cloth for 3 hours. After 3 hours, media is poured into tray. Next, Nata
starter is poured into tray too, then it fermented for 21 days.
In the fifth experiment, the writer try to add yeast. Yeast functioned as a
doer of fermentation. Writers reason of using yeast is because content in
media derived from another carbohydrate so as suppose any saccarification
process. Saccarification process is done minimally by 2 doers of
fermentation, these are Acetobacter xylinum bacteria dan yeast.

Picture 32. Yeast

The fifth experiment was success, but not perfect. Layer is like a gel,
expansible as Nata and smooth on the top, milky colored, and has the
thickness less 1 cm. However, some layer get contamination/broken, it is
because there are another bacteria enter into media. Beside that, newspaper
that used as a cover stick on the layer contaminated.

Picture 33. The Fifth Experiment Result (top view)

26
 As can be seen on the picture above, some layer got contamination, however
some another layer has already form an expansible layer like Nata and white
colored. But, writer is directly to throw away the result to behind of writers home.

Picture 24. The Fifth Experiment Result (Nata layer)

Picture 35. The Fifth Result Experiment (layer is raised)

5. Sixth Experiment
In the sixth experiment conducted on 11 September 2013, the writer did not
use soursop juice as in the previous experiment, but the writer try to use one of
the beverage products such as soursop juice (500 mL). This was done to
compare the ability of the natural soursop juice and soursop juice product
processed by the industry to serve as a medium of Nata. In this case, it means
the writer did not do the filtering.

27
 Picture 36. Beverage of Soursop Mixture Sunglap Juice
Soursop juice (500 mL) was directly mixed with 100 mL of bean sprout
extract, 10 mL of acetic acid, 20 mL of Nata starter, and 50 g of sugar, then
boiled (in this experiment, the writer has not tried to use baker's yeast). After
boil up, media is cooled by letting up to 15 minutes while covered it with a dust-
cloth. Furthermore, the media poured into the tray, and then given a 20 mL of
Nata starter. Close the tray with newspaper, then tie at the both ends with a
rubbers. The final step of this process is save the tray in. After 17 days, open
the tray and observe the results .
The result of the sixth experiment was failed. Media did not form Nata, it
is still liquid, blackish brown colored, and cause a bad odor. In some parts of
the media, there are black spots, as can be seen on the picture below.

Picture 31. The Sixth Experiment Result

Failure caused by antibacterial substance who contained in the product
mixture of soursop beverage, other than that the composition of additives such as
fruit concentrates pear, lychee fruit concentrate, acidity regulator citric acid and
sodium citrate, salt, and vegetable stabilizer. As for the composition of the

28
 chemicals in the beverage products including 43 g of total carbohydrates, 2 g of
dietary fiber, 17 g of sugar, 85 mg of sodium and 150 mg of potassium .
6. Seventh Experiment
In the seventh experiment conducted on 7 October 2013, the writer use the
same materials with previous experiments (1, 2, 3, 4, 5). Such as 500 mL of
soursop juice, 125 mL of bean sprout extract, 20 mL of Nata starter, 75 g of sugar,
15 mL of acetic acid, and 5 g of yeast are mixed then boil it. After that, media is
cooled by letting it in stewpan then it is covered with dust-cloth as at the picture on
the next page.

Picture 32. Cooling Media to the Room Temperature

7. Eighth Experiment
In the eighth experiment conducted on 22 October 2013, the writer use
materials such as 500 mL of soursop juice, 125 mLof bean sprout extract, 20
mL of acetic acid, 100 g of sugar, and 0.25 g of yeast. The materials are mixed
and then boiled. After that, cool the media for 2 hours by letting it in the
stewpan while covered with a dust-cloth. Next the media poured into the tray,
then pour Nata starter into media. The final step of this process is save the
media in cupboard. Writer has tried to be more sterile to the place for
fermentation by soaking more of alcohol and not using the same place, but use
another place.
Fermentation process is not running as well appropriate with the proper
time length. Media is only fermented for 5 days because when writer observe
the condition of tray in the saving room, writer found many insects walk to
approach to the tray. Writer has kept the cleanliness of the place by spraying
alcohol around the place, but insect like ants still come. Ants enter into tray
through small spaces at newspaper until the ants are floating on the media.

29
 This case cause the fermentation was failed totally, because media get
contamination by coming of ants that enter into the tray.

CHAPTER V. CONCLUSION AND SUGGESTION

5.1 Conclusion
Based on the eight times experiments writer did, it can be conclude that
soursop can not be used perfectly as media in making Nata, because media of
soursop easier to get contamination with others bacteria. The nutrient content of
soursop is lowest/highest than nutrient content of coconut water so as makes
bacteria can not grow maximally. Bacteria will grow maximally at the optimum
condition.
5.2 Suggestion
Based on the eight times of experiments writer did, there is suggestion to
be known as the failing happened by writer frequently, that is about cleanliness
of tools used. It is pivotal to do the sterilization of the tools will be used and
always observe the place that used to do the fermentation.

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 BIBLIOGRAPHY

Haryanto, Sugeng. 2009. Ensiklopedi Tanaman Obat Indonesia. PALMALL:

Yogyakarta.

Pradipta, Gabrielle Nindya K. 2011. Buah dan Sayuran Sirsak. Semarang: Makalah
Fakultas Kedokteran Program Studi Ilmu Gizi Universitas Diponegoro.

Sunarjono, Hendro. 1987. Ilmu Produksi Tanaman Buah-Buahan.

Bandung: C.V. Sinar Baru.

Syamsuri, Istamar. 2006. Buku Kerja Ilmiah Biologi SMP 3B. Erlangga: Jakarta.

Yatim, Wildan. 1999. Kamus Biologi. Jakarta: Yayasan Obor Indonesia.

Aroni, Hasan. 2013. Sirsak Si Pencegah Kanker. http://redesain.poltekkes-

malang.ac.id/index.php/rumah/detail/artikel/1/249.Diakses pada 05/10/2013

Hana, dr. Abu. 2011. Kandungan, Khasiat, dan Manfaat Daun & Buah Sirsak.
http://kaahil.wordpress.com/2011/04/08/kandungan-khasiat-dan-manfaat-daun-
buah-sirsak-resep-pengobatan-untuk-kanker-prostat-pankreas-dan-paru-paru-
mengobati-sakit-pinggang-bayi-mencretambeien-bisul-anyang-anyanganliver-e/
Diakses pada 07/10/2013

Maria. 2013. Kandungan Nutrisi dan Manfaat Buah Sirsak Untuk Kesehatan.
http://www.makeitaffordable.com/kandungan-nutrisi-dan-manfaat-buah-sirsak-
untuk-kesehatan/. Diakses pada 05/10/2013

31
Munawar, Muh. Taufiq. 2009. Bakteri Nata De Coco.
http://muhtaufiqmunawar.blogspot.com. Diakses pada 21/10/2013

Resie, Elwin P. 2013. Pengolahan Aneka Buah Sirsak.

http://bbppketindan.bppsdmp.deptan.go.id/diklat/seputar-
diklat/diklatkerjasama/160-pengolahan-aneka-buah-sirsak. Diakses pada
05/10/2013

Sihombing. 2011.
http://repository.ipb.ac.id/bitstream/handle/123456789/51342/f11ndi_BAB%20II
%20Tinjauan%20Pustaka.pdf. Diakses pada 23/05/2013

Stevani, Gusti. 2012. Nata De Coco.

http://verarbeitung2012.wordpress.com/2012/03/11/nata-de-coco/. Diakses pada
07/10/2013

Suprihatin.2010. Teknologi Perpindahan Massa Dalam Perancangan Proses Reaksi.

http://eprints.upnjatim.ac.id/3161/2/fermentasi.pdf. Diakses pada 07/10/2013

Wardanu, Adha Panca. 2011. Bakteri Pembentuk Nata.

http://apwardhanu.wordpress.com/. Diakses pada 07/10/2011

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