report starter.docx

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RATIFICATION PAGE

The complete report of Biotechnology experiment with the title Making

Starter of Natta De Coco created by:

Name : Nur Fahmi Utami

Reg. num : 101 404 155

Class : ICP

Group : One (I)

After checked by Assistant and Coordinator Assistant, so this report has

accepted.

Makassar, December 6th

2013

Assistant Coordinator Assistant

Rachmayani Ardiansyah, S.Pd Muhammad Nur Arsyad

Known by

Responsibility of Lecturer

Prof. Ir. Hj. Yusminah Hala, M.SNIP: 1961 1212 1986 01 1 002


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CHAPTER I

INTRODUCTION

A. BackgroundExperts in United States anticipate the worlds population in 2050 to be

approximately 8.7 billion persons. The worlds population is growing, but its

surface area is not. Compounding the effects of population growth is the fact that

most of the earths ideal farming land is already being utilized. To avoid

damaging environmentally sensitive areas, such as rain forests, we need to

increase crop yields for land currently in use. By increasing crop yields, through

the use of biotechnology the constant need to clear more land for growing food is

reduced

Biotechnology holds the key to increasing the yield of staple crops by

allowing farmers to reap bigger harvests from currently cultivated land, while

preserving the lands ability to support continued farming. Malnutrition in

underdeveloped countries is also being combated with biotechnology. Survey

research over the past decade shows that biotechnology is not likely to become animportant issue for most American consumers. Consumers find biotechnology

acceptable whenthey believe it offers benefits and it is safe. Surveys have

consistently found that a majority of American consumers are willing to buy

insect-protected food crops developed through biotechnology that use fewer

chemical pesticides, as well as more nutritious foods. American consumers also

appreciate the role that biotechnology can play in feeding the world. Research

shows that European consumers are much less supportive of all biotechnology

applications.

Research shows that consumers will accept biotech foods if they see a

benefit to themselves or society and if the price is right. Their responses to foods

developed through biotechnology are basically the same as for any other food -


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taste, nutrition, price, safety and convenience are the major factors that influence

our decisions about which foods to eat.

Therefore one of the products by using biotechnology in particular the

principles of conventional biotechnology is nata de coco fermentation using

Acetobacter xylinumstarter

B. PurposeThe purpose this experiment is to know the steps are making Starter of Nata

De Coco

C. BenefitThe benefit this experiment is the student university can be know the steps

are making starter of Nata de Coco

CHAPTER IIPREVIEW OF LITHERATURE

Nata is from Spayol language that means cream. So, Nata de Cocois cream

from coconut water. That cream is formed from fermentation result of organism


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namelyAcetobacter xylinumthat form gel on the surface of theNata de Cocothat has

sugar (Hala, 2013).

Nata de coco is one of several coconut water potential that most developed in

Indonesia. Nata de coco is a result of coconut water fermentation by using the

Acetobacter xylinum bacteria. Chemically, the fiber contained in nata de coco is a

cellulose fiber, known as bacterial cellulose [Piluharto, B: Saputra and Darmansyah,

2010]. Bacterial celluloses have some advantages such as having a high purity

without lignin, pectin, and hemicelluloses, which are commonly found in plant

cellulose [S, Makoto et al. : Saputra and Darmansyah, 2010].

Acetobacter xylinum bacteria including gram- negative, rod-shaped,

microaerophilic and are catalase positive, this bacterium belongs to a group of acetic

acid bacteria through the process of oxidation of methyl alcohol to produce acetic

acid. Acetic acid is what serves as a growth suppressor acidophilic. Acetobacter

xylinumcannot produce amylase but can produce specific disakaridase like sucrase.

Acetobacter xylinum is not pathogenic in humans and animals (Budiyanto, 2002).

Besides these bacteria also produce enzymes that make up (polymerize)

polymerization of glucose to compound known as extracellular cellulose nata

(Pambayun: Budiarti, 2010).

Formation of extracellular cellulose synthesis Acetobacter xylinumoutcome

is the result of the conversion of sugar and other carbon sources. Nata formation

occurs because the process of making glucose from sugar solution or medium

containing glucose by cells ofAcetobacter xylinum . The glucose combined with fatty

acids to form precursors to the cell membrane. Precursor is issued in the form of

excretion and with polymerizing glucose to cellulose enzymes outside the cell

(Pambayun: Budiarti, 2010).

According Pambayun in Budiartis Journal, 2010,Acetobacter xylinumwill

pass through several phases of growth as follows:

a) Phase adaptationb) Early Growth Phase


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c) Exponential Growth Phased) Slow Growth Phasee) Fixed Growth Phasef) Phase Towards Death

Acetobacter is a bacteria that produces cellulose fibers are very delicate.

These fibers can form a network on the surface layer between air and liquid called

pellicle. The pellicle has a thickness of approximately 10 mm dependent on microbial

growth period. Pellicle which is located on the surface of the liquid air made up of

ribbons containing crystalline high. The ribbons have a width of 40-100 nm , but the

length is difficult to measure because the network form relating to one another . The

ribbon is made up of parts of microfibrils associated through hydrogen bonding

(Figini: Anam, 2010).

The cellulose - containing materials usually form a crystalline structure , so

that water cannot enter the stricken active at temperatures of crystalline bacterial

rooms. Cellulose has different characteristics in its crystalline structure. Cellulose

contains two different crystalline structures items, namely cellulose and cellulose 1

1. On cellulose 1, one triclinic unit cell contains one cellulose chain, while the

cellulose 1 one monoclinic unit cell contains two cellulose chains. Bacterial

cellulose - containing cellulose 1 approximately 60 %, this is different from

cellulose derived from plants (eg flax and cotton) containing cellulose 1 only 30 % ,

whereas cellulose compensates 1 ( Yoshinaga et al : Anam , 2010) .

Seeds for nata can be obtained freely from nature. Fruits

containing sugar and vegetables that had rotted a media (where growth) is good for

the bacteria Acetobacter xylinum. The filtrate pineapple is often used as a growth

medium for making pure cultures of capturing nature. This method is relatively

inexpensive and requires tools simple (Wahyudi, 2003).

The starter is the seed Acetobacter xylinum which had been grown in a

substrate the culture that population growth Acetobacter xylinum achieve optimal

density for making nata making, i.e. 1 x 109 cells / ml. Usually this will be achieved


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density on the growth of the culture in substrate for 48 hours (2 days). Preparation of

starter is as follows: sterilized substrate autoclave or by boil for 15 minutes. After

cold milk approximately 40 C, 300 ml was added to the volume of 500 ml sterile

bottles. Substrate inoculated in sterile bottles (A. xylinumbacteria seedlings planted)

by 2 loops (approximately 2 bulb matches), seedlings A. xylinum. Substrate shaken

out preferably uses a shaker with a speed of 140 rpm (manually shaken every 2-4

hours). Starter grown for 2 days at room temperature (Misgiyarta, 2007).

CHAPTER III

EXPERIMENT METHOD

A. Time and PlaceDay/Date : Thursday/28

thNovember 2013

Time : 11.00 sd 13.00 CIT


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Place : Experimentoratory of Biology Department of Mathematics

and Science Faculty Macassar State University

B. Tool and MaterialTools :

1. Stove2. Chamfer3. Pot4. Newspaper5. Rope6. Spoon7. Filter8. SyringeMaterials :

1. Coconut water 5 L2. Glacial vinegar (Asetat acid 100%) 12 mL3. Sugar 8 gr4. Starter (bakteriAcetobacter xylinum)5. Fertilizer ZA 8 gr6. Ph Meter (pH paper)

C. Work procedure1. Filtered coconut water then cook until boiling. After boiled let it for 10

minutes

2. Added other ingredients and let boiled for 10 minutes3. Checked for up to pH 3-44. Fill into the gutters that have been sterilized beforehand, closed with the first

newspaper in alcohol spray. Stored in a safe place, save for a day (24 hours).


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5. Poured starter (bacteria Acetobacter xylinum) into the gutters, store in a safeplace.

6. Wait for 5-7 days, then nata ready for harvest

CHAPTER IV

OBSERVATION RESULT

A. Observation ResultTable of lateral bud lenght and stem diameter


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Starter

B. Discussion


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Based on observations obtained, this experiment was success.

Reflecting the changing coconut water and some coconut water did not change

after being stored for 6 days. Changes can be seen that there is a pile of

compaction or white on the surface of coconut water. This proves that the

fermentation of coconut water with the help of bacteria Acetobacter xylinum.

Sugar in the coconut water is converted to acetic acid and cellulose threads. Over

time will form a solid mass and reaches a thickness of a few centimeters. Thus,

nata de coco can also be considered as a form of bacterial cellulose dense, white,

transparent, sweet taste and chewy texture.

Carbon source is an important factor in the fermentation process. Bacteria

needs it to make nata for metabolism process, glucose to enter the cells and used

for energy supply that needed for breeding, fructose that there will be synthesized

into cellulose. The amount of sugars adding should be noted that sufficient for

metabolism and pellicle formation nata. Although there is sugar in the coconut

water but the sugar is not sufficient to form pellicle, so that need to add from the

outside.

Nutrients contained in coconut water include: sugar sucrose 1.28%,

mineral resources, such as; Mg2 +3.54 g / l), as well as the factors supporting

growth (growth promoting factor) is a compound that is capable of enhancing the

growth of bacteria nata (A. xylinum). The presence of sucrose sugar in the

coconut water will be utilized by A. xylinum as a source of energy, as well as a

carbon source to form the metabolites of which is cellulose which forms Nata de

Coco. Compounds enhancing microbial growth (growth promoting factor) will

increase the growth of microbes, whereas the presence of minerals in the

substrate will help to increase the activity of the kinase enzyme in the

metabolism of the cell in A. xylinum to produce cellulose.

Based on the experiment result, we could see the function of all

materials that we used, they are :


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1. Glacial vinegar (Acetate acid)Acetic acid were added in coconut water serves to reduce or increase the

degree of acidity. Type of vinegar is best qualified to produce nata is glacial

acetic acid with a concentration of 99.8% acidity.

2. Granulated sugarThe addition of sugar that serves as a carbon and energy source of the bacteria

that play a role in the production of nata de Coco.

3. ZA fertilizerZA function is as a source of nitrogen, so that the bacteria Acetobacter

xylinum can live and thrive. The content of ZA is Ammonium Sulfate. ZA

also as a medium for bacteria Autotrophs.

4. Acetobacter XylinumFunctions convert the sugar in the coconut water into sheets of cellulose

fibers. Cellulose sheets were then becomes solid and translucent white, called

nata.

CHAPTER VCLOSING

A. ConclusionThe bacteria are put into the starter Acetobacter xylinum.Factors that affect

the growth of Acetobacter xylinumare nutrients, carbon source, nitrogen source,

medium, acidity, temperature, and air (oxygen). This proves that the fermentation

of coconut water with the help of bacteria Acetobacter xylinum. Sugar in the

coconut water is converted to acetic acid and cellulose threads. Over time will

form a solid mass and reaches a thickness of a few centimeters. Thus, nata de

coco can also be considered as a form of bacterial cellulose dense, white,

transparent, sweet taste and chewy texture.


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B. Suggestion1. Laboratory

Prepare the fulfill tools and materials, so the experiment is success.

2. AssistantAssistants have to guide the apprentice when doing experiment until it finish,

so the apprentice not confused or panic when something happens in

experiment.

3. ApprenticeApprentices have to serious when doing experiment and notice the

experiment procedures also every apprentice has to make a temporary report.

BIBLIOGRAPHY

Anam. 2010. Mikrobiologi Umum.cet. kedua. UMM Press. Malang.

Budiarti. 2010.Biologi Edisi Kedelapan Jilid 1. Jakarta: Erlangga.

Hala, Yusminah. 2013. Pengantar Bioteknologi. Makassar: Jurusan Biologi FMIPA

UNM.

Misgiyarta, 2007.Research Progress in Production of Bacterial Cellulose by

Aeration and Agitation Culture and Its Application as a New Industrial

Material. Biosci. Biotech. Biochem., 61:219224.

Wahyudi. 2003. Bioremediasi Limbah Cair Nanas Sebagai Bahan Baku Pembuatan

Nata De Pina. Malang: FMIPA UM.


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