Theme: Environmental Conservation

Title: Converting garbage waste into cleaning agent

Prepared by : Kayalvilli a/p Shanmugam : Tharuchanna a/p Subramaniam : Divvya a/p Sivarajah : Nur Iman bt. Mohd Isa : Ummi Ezzatty bt. Bahari : Kishen a/l Nanda Kumar : Chong Swee Hong : Felicia Neoh Yee Peih

ABSTRACT Garbage Enzyme is an organic solution produced by the simple fermentation of kitchen waste, brown sugar and water, in much the same process that wine is made. This solution has the capacity to breakdown, change, create and catalyse, therefore making it a good household cleaning aid. It is simple and inexpensive. We made our garbage enzyme by collecting kitchen wastes from the school canteen, recycling them and fermented it together with brown sugar and water in containers for three months. Unlike most commercial detergents which cause water pollution and subsequently affect the environment adversely, garbage enzyme is more environmentally-friendly and reduces pollution. Therefore, garbage enzyme is a much better alternative to chemical household cleaners. Our objective of this project is to find out the chemical properties of garbage enzymes, and to establish the ability of garbage enzyme as a stain remover and disinfectant. This was done by using distillation method, removal of various types of common stains from fabric and observing of the effect of enzymes on bacterial colony. The main findings of our project are that garbage enzymes are acidic in nature, most effective as stain remover when used with soap and clearly show antibacterial property. Additionally, we hope to inform more people that garbage enzymes can be as effective as chemical detergents and encourage more widespread use of garbage enzymes. Our mission is to reduce the amount of soap used in our daily application and instead, replace half the amount of soap needed with the usage of garbage enzyme.

LIST OF CONTENT

NO

TITLE

PAGES

ABSTRACT 2

LIST OF CONTENTS

INTRODUCTION

45

THEORETICAL BACKGROUND

68

METHODOLOGY

9 16

RESULTS AND DISCUSSION

17 20

CONCLUSION

21 22

APPENDIX

23 40

REFERENCES / ACKNOWLEDGEMENT

41

INTRODUCTION

This experimental project is about recycling garbage waste which can be obtained from the school canteen freely and converted into cleaning agent known as garbage enzyme. This garbage enzyme is produced from the kitchen waste through the process called fermentation for three months. Garbage enzyme was developed by Dr. Rosukon Poompanvong from Thailand. She has been actively involved in enzyme research for more than 30 years and encourages people to make garbage enzyme at home to ease global warming. According to Dr Rosukon's Garbage Enzyme is a complex organic substance of protein chains , mineral salts and juvenile hormones. The main purpose of this investigation is to examine the properties of various garbage enzyme in hopes of finding a non-synthetic or all-natural cleaner equally well in a broad variety of areas to synthetic ones. These are their antibacterial properties and removal of a variety of stains on fabric. This theme was chosen to foster good environmental values amongst the school community and to raise awareness amongst the school community about the importance of environmental protection and conservation. Besides that, we hope to encourage the school community to carry out effective environmental activities of producing garbage enzyme and to generate a school surrounding that is conducive for producing a school community that practices a sustainable way of life. Normally people will throw away food waste such as fruits scraps and vegetables into the bin. To avoid waste, it can be recycled to produce cleaning agent or enzyme that are multipurpose such as to clean the classroom, the toilet and even as a fertilizer. By producing our own product in school, we can save our budget due to the slow down of world economy. Moreover, this product is eco-friendly without side effect. Detergents can have poisonous effects in all types of aquatic life if they are present in sufficient quantities, and this includes the biodegradable detergents. Most fish will die when detergent concentrations approach 15 parts per million. Detergents also add another problem for aquatic life by lowering the surface tension of the water. Organic chemicals such as pesticides and phenols are then much more easily absorbed by the fish. Phosphates in detergents can lead to freshwater algal blooms that release toxins and deplete oxygen in waterways. When the algae decompose, they use up the oxygen available for aquatic life. However, garbage enzyme is non-toxic and antibacterial, and does not harm the environment. Garbage enzymes can be produced manually and easily. The cost of making garbage enzymes is cheap whereas the price of soaps nowadays are quite expensive. It saves

money by turning kitchen waste into multipurpose natural household cleaners. This is an excellent way to do our part in safeguarding the environment. Garbage enzyme is the solution to recover our ozone layer and lower the global temperature. It saves the environment by reducing landfills which directly reduce methane emission. It also acts as a natural pesticide which reduces the demand for chemical pesticides. Therefore, rather then throwing the food wastes just like that, why not convert them into cleaning agent that is ecofriendly, odourless and save money! From our investigation, we hope to prove that garbage waste can be made into effective cleaning agents, hence reducing the usage of soap, detergents and other cleaning chemicals that has negative effect to the environment. It is our mission to heighten cooperation between schools and communities in making environmental education a success. In future, hopefully this project can be implemented by the society because of ecofriendly factor.

THEORETICAL BACKGROUND The scientific concept involved in the conversion of garbage waste into cleaning agent is fermentation. Fermentation is a chemical change brought on by the action of microscopic yeast, moulds and bacteria. Yeast has an enzyme called zymase and this catalyses the fermentation process. Glucose zymase Ethanol + carbon dioxide C6H12O6 (aq) 2C2H5OH(aq) + 2CO2(g) Acetobacter bacteria convert alcohol to acetic acid in the presence of excess oxygen. The oxidation of one mole of ethanol yields one mole each of acetic acid and water; C2H5OH + O2 Alcohol + Oxygen CH3COOH = + H2O + water

acetic acid

Species of anaerobic bacteria, including members of the genus Clostridium, can convert sugars to acetic acid directly, without using ethanol as an intermediate. The overall chemical reaction conducted by these bacteria may be represented as: C6H12O6 3 CH3COOH Glucose = acetic acid

To understand the product formed during fermentation, some background information on the type of microorganism and enzymes were gathered. Bacteria Several bacterial families are present in foods, the majority of which are concerned with food spoilage. As a result, the important role of bacteria in the fermentation of foods is often overlooked. The most important bacteria in desirable food fermentations are the lactobacillaceae which have the ability to produce lactic acid from carbohydrates. Other

important bacteria, especially in the fermentation of fruits and vegetables, are the acetic acid producing acetobacter species.

Yeasts Yeasts and yeast-like fungi are widely distributed in nature. They are present in orchards and vineyards, in the air, the soil and in the intestinal tract of animals. Like bacteria and moulds, yeasts can have beneficial and non-beneficial effects in foods. The most beneficial yeasts in terms of desirable food fermentation are from theSaccharomyces family, especially S. cerevisiae. Yeasts are unicellular organisms that reproduce asexually by budding. In general, yeasts are larger than most bacteria. Yeasts play an important role in the food industry as they produce enzymes that favour desirable chemical reactions such as the leavening of bread and the production of alcohol and invert sugar. Moulds Moulds are also important organisms in the food industry, both as spoilers and preservers of foods. Certain moulds produce undesirable toxins and contribute to the spoilage of foods. The Aspergillus species are often responsible for undesirable changes in foods. These moulds are frequently found in foods and can tolerate high concentrations of salt and sugar. However, others impart characteristic flavours to foods and others produce enzymes, such as amylase for bread making. Moulds from the genus Penicillium are associated with the ripening and flavour of cheeses. Moulds are aerobic and therefore require oxygen for growth. They also have the greatest array of enzymes, and can colonise and grow on most types of food. Moulds do not play a significant role in the desirable fermentation of fruit and vegetable products. During fermentation,micro-organisms metabolise and grow they release by-products. In food fermentations the by-products play a beneficial role in preserving and changing the texture and flavour of the food substrate. For example, acetic acid is the by-product of the fermentations of some fruits. Enzymes The changes that occur during fermentation of foods are the result of enzymic activity. Enzymes are complex proteins produced by living cells to carry out specific biochemical

reactions. They are known as catalysts since their role is to initiate and control reactions, rather than being used in a reaction. Because they are proteinaceous in nature, they are sensitive to fluctuations in temperature, pH, moisture content, ionic strength and concentrations of substrate and inhibitors. In the food industry, enzymes have several roles the liquefaction and saccharification of starch, the conversion of sugars and the modification of proteins. Microbial enzymes play a role in the fermentation of fruits and vegetables. Nearly all food fermentations are the result of more than one micro-organism, either working together or in a sequence. For example, vinegar production is a joint effort between yeast and acetic acid forming bacteria. The yeast convert sugars to alcohol, which is the substrate required by the acetobacter to produce acetic acid. In general, growth will be initiated by bacteria, followed by yeasts and then moulds.

METHODOLOGY

Various methodology were used in our investigation as shown below:

1. Statistic This method was used to gather information on the amount of food waste produced in the school canteen and hostel for a month.

2. Synthesis This method was used to produce the cleaning agent from garbage waste which is a by-product of the fermentation process.

3. Experimental method This method was carried out by carefully planning the experimental procedures, the apparatus and materials, controlling the variables and obtaining and analysing the data collected. The aim of the experimental method is to study the properties of the garbage waste produced and to prove that the garbage waste produced are effective as cleaning agents in terms of removing stains and as a disinfectant.

4. Interview We carried out interviews with teachers (housewives) to hear about their experiences in making garbage enzymes. The good point is that the information collected is first hand information and from people who are experienced in this area, thus making our research more reliable.

Statistic

The waste products collected from the school canteen were weighed and the data were recorded. The data collected were then transferred into graph.

BAR GRAPH SHOWING THE WASTE COLLECTED IN GRAMS WITHIN THE PERIOD OF FOUR WEEKS.
2000 1800 1600 1400 1200 1000 800 600 400 200 0 First week Second week Third week Fourth week Sunday Monday Tuesday Wednesday Thursday

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Synthesis The steps to produce garbage enzyme are as follows: 1. Kitchen waste which were collected from the school canteen and hostel were separated by discarding oily food and using only fruits and vegetables. 2. Then, 200g of brown sugar, 600g of waste (fruits and vegetables) and 2000g of water were mixed well in a 3 litre plastic bottle. (Ratio of sugar: waste: water = 1: 3: 10) 3. The bottles were closed tightly. 4. The mixtures in the bottles were fermented for three months. 5. Steps 1 to 4 were repeated using fruits only and vegetables only to produce different types of garbage enzyme products. 6. The bottles were kept in a cool dry place at room temperature ( 27C). 7. The bottles were opened once a day for the first month to release excess gas produced during fermentation and to prevent the bottles from expanding. The bottles were re-capped tightly each time after the gas was released. 8. For the second and third months, the gases were released when necessary. 9. Every now and then, the mixtures were given a good stir so that it gets enough air. 10. If there are formation of white layer (yeast) during fermentation, a handful of brown sugar were added and they will disappear overnight.

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11. After three months, the enzymes were filtered and poured into different bottles. The residues were dried, then blended and buried in the ground as a fertilizer.

Experimental Procedure 1: Problem Statement: What is the property of the product of fermentation (garbage enzyme)? Aim: To investigate the properties of garbage enzyme Hypothesis: Garbage enzymes are by-products of fermentation Variables: Manipulated: Types of garbage enzymes Responding: The pH, appearance and smell of the fractions of distillation collected Fixed: Amount of garbage enzyme, distillation temperature Materials: Garbage enzyme, pH meter Apparatus: Boiling flask, Claisen adaptor, condenser, vacuum adaptor, conical flasks, retort stand, Bunsen burner, tripod stand, thermometer.

The process of distillation

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Procedure: 1. The distillation apparatus were prepared as shown in the diagram above. 2. The garbage enzyme were filtered into the distillation flask. Fractions were collected between 77100C. 3. The appearance , odour and pH of the distillate was recorded. 4. The flammability of the distillate obtained was tested.

Experimental Procedure 2: Problem Statement : How does different types of enzymes react on different types of stains? Aim : To investigate the effectives of different types of enzymes on different types of stains. Hypothesis : The ability to remove different types of stains depends on the effectiveness of different types on enzymes. Variables : Manipulated : Types of enzymes , Types of stains Responding : Ability of enzymes to remove stains Constant : Amount of water used , Amount of enzymes used , Time taken to dry the clothes. Material : Water, Chili sauce, ink, curry, coffee, fruit enzymes, vegetable enzyme, fruit and vegetable enzyme, soap powder. Apparatus : Pieces of cloths, glass containers, glass rods, beakers, dropper, spatula, forceps, thread, bottles, measuring cylinder, retort stand, basin, plastic jug, labeling stickers, white tiles.

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Procedure: 1. 50 ml of different enzymes were measured and poured into their respective beakers. 2. The beakers were labelled according to the type of enzymes. 3. Four beakers containing 50ml of water and 50ml of concentrated soap solution respectively was prepared as control. 4. Then, several cloths were cut into identical sizes to be stained. 5. The cloths were stained separately with curry, chilli sauce, coffee and marker ink to be tested with different types of enzymes. 6. The stained cloths were left to dry. Once dried, the stained clothes, were engrossed into the beakers filled with enzymes, water and soap. 7. Then the beakers were stirred using different glass rods for each enzymes. 8. The cloths were immersed in the beakers containing the enzymes for an hour. After an hour, the cloths were removed from the beakers and rinsed in a basin. 9. The cloths then were left to dry and labelled. 10. Our observation was recorded in a table. 11. Steps 1 to 7 were repeated using enzymes diluted with water and enzymes diluted with soap solution with the ratio of 1 : 1 with the total volume of 50ml.

Experimental Procedure 3: Aim : To investigate the antibacterial property of garbage enzyme. Hypothesis: Garbage enzymes are more effective as disinfectant( has antibacterial properties) compared to soap Materials: Nutrient agar , garbage enzyme , soap solution , saliva samples. Apparatus: Petri dishes , cotton swab , filter paper , puncher , beaker , forceps , cellophane tape , marker pen. Variables: Manipulated : Type of solution (soap and enzyme) Responding : Indication of clear area around the disc (antibacterial property) Fixed : Type of bacteria colony, temperature, nutrient

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Procedure: 1. Three petri dishes with nutrient agar were prepared. 2. A sample of saliva was collected from one of the students. 3. Cotton swab was dabbed into the saliva and was swabbed across the surface of the agar. 4. The petri dishes were sealed using cellophane tape to avoid contamination. 5. The petri dishes were left in a dark cupboard for two days to allow the bacterial growth. 6. After two days, filter papers were punched using puncher into small discs. 7. The bacterial colony (white spots) formed were circled using marker pen on the bottom of the petri dishes. 8. Then, the discs were immersed into the garbage enzymes solution and soap solution which were placed onto the bacterial colony formed on the agar in different petri dishes respectively. 9. The petri dishes were labelled and left in the cupboard for two days. 10. After two days, observation was recorded.

Flow Chart of the Experiment Kitchen wastes were collected produced by using and weighed. A complex solution was

kitchen wastes, organic substances, sugar (brown

sugar, jiggery or molasses sugar) and water. The solution was fermented for three months in separate containers.

Once a day for the first month, the containers were opened regularly to release the ozone gases formed to avoid the increasing of carbon dioxide in the air. The containers were re-capped tightly each time after the gases were released. When there was a white layer on the surface of the enzyme solution, a handful of sugar was added and mixed well.

After three months, the product was filtered and separated into dissimilar labelled bottles. ( Fruit enzyme, Vegetable enzyme, Fruit and vegetable enzyme, and Papaya leave enzyme). The characteristics and constituents of the enzyme solution were investigated.

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Experiments were conducted to verify the effectiveness of different types of enzymes towards different types of stains. The results obtained were tabulated. The observations were recorded.

To make the cleaning agents (enzymes) more effective, the experiment was repeated using soap produced through saponification process.

Conclusions were made based on the results obtained.

Implementing the enzymes as a cleaning agent in classrooms, school toilets, canteens and staff rooms. Promoting garbage enzyme as a cleaning agent and its usage to the school teachers and students.

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RESULTS AND DISCUSSION:

Experimental Procedure 1: Observation:

1. Distillate collected was a clear liquid. 2. The distillate smelt like vinegar. 3. The pH of the distillate was 3.8. 4. This fraction burned easily compared with the non-flammable original solution.

The process of condensation was carried out to determine the properties of the enzyme. The distillate was collected in a round bottom flask. The product obtained was colourless. The solution was then inhaled from a distance. Make sure the liquid was not inhaled directly to avoid the gas from affecting our brain cells. It smelt sour which proves that it has an acidic properties. To prove the acidity of the distillate, it was tested using pH meter. The fraction obtained was partially acetic acid which was causing it to show its acidic properties. This makes it to smell like vinegar. Through this investigation, we can conclude that half the amount of soap can be reduced by replacing it with the usage of garbage enzyme. The fraction obtained contain ethanol causing it to be highly flammable. Refer to the data compiled in appendix for further information.

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Experimental Procedure 2: GARBAGE ENZYME TABULATION OF DATA Enzyme Type of stains Concentrated Enzyme + Water 1 2 3 1 2 3 1 Enzyme + Soap 2 3

A Curry

-S -W

-S -W

-S -W

-S -W

-S -W

-S -W

+S +W

+S +W

+S +W

B Chilli Sauce

+S +W

+S +W

+S +W

+S +W

+S +W

+S +W

-S -W

+S +W

+S +W

C Coffee

-S -W

-S -W

-S -W

-S -W

-S -W

+S +W

+S +W

+S +W

+S +W

D Marker Ink 1 Fruit Enzyme

-S -W

-S -W

+S +W

-S -W

-S -W

-S -W

+S +W

-S -W

+S +W

2 Vegetable enzyme

+S : Effective than soap

+W : Effective than water.

3 Fruit and Vegetable enzyme

S : Less effective than soap

W : Less effective than water.

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The results obtained were compared with the control experiment (water and soap). According to the data collected, the three concentrated enzymes were not effective than water and soap when they were tested with different types of stains. Eventhough the concentrated enzymes removed the stains partially but it ended up staining the cloths which make the white cloths to turn brown in colour. This shows that the concentrated enzymes are not very effective to be used. Secondly , the three enzymes were diluted with water with the ratio of 1 : 1 ( 25 ml of enzyme and 25 ml of water ). This solution was used to test with the different type of stains. Based on the by product obtained , we can conclude that this enzyme solution can remove the stains moderately which is much better than the concentrated enzymes. Next , the different types of the enzymes were diluted with soap solution with the ratio of 1 : 1 ( 25 ml of enzyme and 25 ml of soap solution ). This solution were also tested with the stains prepared. The results earned shows that this solution managed to remove almost all the stains completely. Besides , the enzyme diluted with soap solution is the best compared to concentrated enzyme and enzyme diluted with water. More specifically , fruit and vegetable enzyme diluted with soap solution is the most effective solution than the soap and water. As a conclusion , we can summarize that the soap alone cannot digest the stains. It needs the aid of enzymes to fasten the reaction of removal of the stains. This is because the stains can develop a resistance against the soap. However, when the solution of enzymes diluted with soap were used, the stains were able to be removed.

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Experimental Procedure 3 :

Type of solution Garbage enzyme

Observation Clear area was observe around the filter paper disc No observable changes

Soap

The experiment was conducted to identify the antibacterial property of the enzyme. Bacteria was produced using saliva which contains more than 10 000 of strains of bacteria. The observation shows clear disc area around the filter paper disc. This is because the antibacterial property of the enzyme causes the bacteria to grow away from the spot stained with the enzyme.The higher levels of acid inhibit the growth of bacteria. So, this investigations proves that the enzyme is an acid which inhibits bacterial growth. Those harmful microbacteria were broken down by the enzyme. The pH affects the ionization and the binding and interaction of many different kind of molecular process which in turn destroys the bacteria. Besides that, garbage enzyme contains ethanol which is a dehydrating agent leaving the surface dry and this can destroy the cell wall of microorganisms, causing the inner contents to flow out leading to cell death. It's used in labs as a sterilizing agent.

Whereas when using soap which is alkaline, it doesnt destroy the bacteria. This is because although raising the pH level of bacteria may kill them, this does not necessarily result in preventing bacteria from growing in any particular environment compared to the acidic medium. Soap cannot remove the bacterial cell unlike the acidic enzyme. Soap isn't as powerfull as ethanol for sterilizing purpose because bacteria may develop a resistance to the alkaline soap. Furthermore, the soap sample used was taken from the school janitor which was used to clean school toilets. In this case, enzyme is needed to fasten the reaction of the removal of stains more effectively.

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CONCLUSION

The process of distillation which was carried out on the enzyme revealed several properties. The garbage enzyme consists of a mixture of ethanol and acetic acid. The presence of the acetic acid makes the mixture acidic and has the smell of vinegar. The presence of ethanol makes the fraction flammable. In the experiment to examine the effectiveness of stain removal, the concentrated enzymes managed to remove the stains partially but the enzymes ended up staining the cloths. Whereas, when diluted enzyme was used, it managed to remove the stains better than concentrated enzyme without staining the cloth. Finally in the experiment using enzyme diluted with soap solution, the results obtained were the best. The enzymes diluted with soap solution managed to remove the stains effectively and the enzymes did not stain the cloth. In the experiment to examine the antibacterial property of the enzyme, it was found that a clear area has formed around the disc saturated with enzyme whereas soap solution did not show any conclusive evidence of antibacterial property. It is proven conclusively that enzymes are effective as disinfectant. The acidic property of garbage enzyme enables it to act as a cleaning agent by dissolving, catalysing and breaking down the dirt and stain. The garbage enzymes acidity could also be a contributing factor for its antibacterial property allowing it to destroy the bacteria formed on surfaces. Our investigation has proven that enzymes can be used in various situations. Knowing the effectiveness in removing stains and its antibacterial property, we can apply the usage of enzymes by cleaning the air filters in the air-conditioners. Besides that, it is used to disinfect things exposed to human contact. For instance, office telephones, door knobs, toilet bowls and sinks, laboratory tables and apparatus, classroom tables, computer keyboards and so on. Besides that, our hostel tenants could use the enzyme as detergent to wash their clothes. In addition, the enzyme helps to deodorise the air by releasing its natural fresh smell as well as dissolving the toxic air released. This helps in using it as an air freshener in the staff room and office. Its smell also drives away insects and pests.

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Our investigation will subsequently reduce the usage of chemical cleaning agents and soap which has adverse effect to the environment. Furthermore, the cost for maintaining the cleanliness in school will be reduced. The garbage in school will also be reduced and thus reducing the amount of organic waste in landfills. As we know, organic waste in landfills will pose adverse effect to the environment by releasing methane gas. Our team would like to extend our investigation by finding an enzyme that is as or more effective than soap which can replace soap altogether. We would like to experiment making enzyme from common herbs like aloe Vera, papaya leaves, tea leaves and so on. Also we would like to investigate the effectiveness of enzyme to digest oil spillage so that it can be used to clear up sea water contaminated with oil.

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APPENDIX EXPERIMENT 1 INVESTIGATE THE PROPERTIES OF ENZYME

The apparatus was set up as shown.

The enzymes were distilled and the product was collected.

23

The product was collected and tested with pH meter.

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EXPERIMENT 2 COMPARING THE EFFECTIVENESS OF VARIOUS ENZYMES ON DIFFERENT STAINS

The garbage waste was collected from the school canteen.

The garbage waste was cleaned and separated according to different categories. For example, fruits and vegetables. Then, it was fermented for 3 months.

The enzymes were stirred using glass rods.

25

The enzymes were filtered

The filtered enzyme was poured into the bottle.

The bottles were labelled according to the types of enzymes.

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Different types of stains were prepared.

The clothes were stained

Stained clothes were left to dry for.

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The enzymes were measured and poured into the beakers.

The beakers containing the enzymes were labelled.

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Soap solution and water were set up as the control experiment.

The clothes were soaked for an hour.

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The clothes were ready to be rinsed in water.

The clothes were dried after washing.

The clothes were labelled.

The results were compared.

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Collected data was recorded

EXPERIMENT 3 EXAMINE THE ANTIBACTERIAL PROPERTY OF ENZYMES COMPARED TO SOAP

Saliva was collected.

The saliva was put on the agar using a cotton bath

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The petri dishes were sealed using cellophane tape.

The petri dishes were left for 1 week to allow the growth of bacteria.

The bacteria spots were circled with marker to emphasis the presence of bacteria.

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Small pieces of filter paper were soaked in the enzyme and placed on the growing bacteria.

The petri dishes were labelled according to their cleaning agents (enzymes).

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The white spots indicate the spots of bacteria that was stained with soap, while the single spot that is clear indicate the bacteria spot that was stained with enzyme

The observation shows that enzyme has antibacterial properties. This is because the filter paper saturated with enzyme is clear from enzyme indicating that the bacteria is moving away from the enzyme.

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Table 1: Micro-organisms commonly found in fermenting fruit and vegetables Organism Type Optimum conditions aw > =0.9 Reactions

Acetobacter genus A. aceti A. pasteurianus A. peroxydans Streptococcaceae Family

Aerobic rods

Oxidise organic compounds (alcohol) to organic acids (acetic acid). Important in vinegar production.

Gram positive cocci

Acid tolerant aw > =0.9 Homofermentative. Most common in dairy fermentations, but S. Faecalis is common in vegetable products. Tolerate salt and can grow in high pH media.

Streptococcus genus S. faecalis S. bovis S. thermophilus

Leuconostoc genus L. mesenteroides L. dextranicum L. paramesenteroides L. oenos

Gram positive cocci

Heterofermentative. Produce lactic acid, plus acetic acid, ethanol and carbon dioxide from glucose. Small bacteria, therefore have an important role in initiating fermentations. L. oenos is often present in wine. It can utilise malic acid and other organic acids. Saprophytic organisms found in fermenting vegetables, mashes, beer and wort. Produce inactive lactic acid.

Pediococcus genus P. cerevisiae P. acidilactici P. pentosaceus Lactobacillaceae Family Gram positive rods. Nonmotile Acid tolerant aw > =0.9

Metabolise sugars to lactic acid, acetic acid, ethyl alcohol and carbon dioxide.

Lactobacillus genus

The genus is split into two

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types homo- and heterofermenters. Saprophytic organisms. Produce greater amounts of acid than the cocci

Homofermentative Lactobacillus spp. L. delbrueckii L. leichmannii L. plantarum L. lactis L. acidophilus Heterofermentative Spp. L. brevis L. fermentum L. buchneri

Produce only lactic acid. L. plantarum important in fruit and vegetable fermentation. Tolerates high salt concentration.

Produce lactic acid (50%) plus acetic acid (25%), ethyl alcohol and carbon dioxide (25%). L. brevis is the most common. Widely distributed in plants and animals. Partially reduces fructose to mannitol.

Yeasts

Tolerate acid, 40% sugar aw > =0.85 Many aerobic, some anaerobes pH 4-4.5 20-30 C S. cerevisiae can shift its metabolism from a fermentative to an oxidative pathway, depending on oxygen availability. Most yeasts produce alcohol and carbon dioxide from sugars.

Saccharomyces Cerevisiae S. pombe

From the background information gathered, we can summarise that the fermentation of garbage waste is a reaction caused by the presence of microorganisms that are found naturally in the food products and the by-products of fermentation depends on the type of microorganism present.

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APPLICATIONS OF ENZYME IN OUR SCHOOL ENVIRONMENT.

The dilute enzyme solutions can be used to clean the keyboards.

The enzyme solution was used to clean the classroom windows.

The dirt from the windows were removed effectively by the enzyme.

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The stain on the staffroom floor

Removal of stain on the floor of the staffroom using the enzyme.

The floor after cleaned with the enzyme solution.

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DILUTION RATE

USAGE Bath (add in bath water to improve skin condition) Washing machine (clean and soften laundry) Toilet bowl (prevent blockage & purify sewage)

QUANTITY

APPLICATION

50-100cc

Keep overnight

20-50cc

Soak and wash

250cc

Pour and wash

Concentrated enzyme

Toilet cistern (purify water)

20-50cc (two caps) 1/10,000 litre water Appropriate amount

2-3 times/ weeks

Garden pond & water tank (purify water)

Add occasionally

Leather sofa (clean mold and stain)

Spray and wipe every 10 days

Carpet & tatami mat (deodorise & antiseptic) Shoe, inside the car (deodorise & antiseptic) Kitchen sink, stove, cooker hood(clean oily stain)

Slightly moist

Spray 1-2 times/month

Appropriate amount Appropriate amount Appropriate amount Appropriate amount Appropriate amount

Spray occasionally

10-50 times

Soak and wipe occasionally Soak and wipe occasionally

Black mold (antifungal)

Pet excretion, pet house (deodorise & antiseptic) Air-cond room (humidifier, deodorise & antiseptic)

Spray occasionally

200 500 times

Spray occasionally

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Bathroom sink (clean)

Appropriate amount Appropriate amount Appropriate amount Appropriate amount Appropriate amount Appropriate amount

Soak and wipe occasionally

Cabinet and refrigerator (deodorise)

Spray occasionally

500 times

Drain (prevent drainpipe blockage)

Flush occasionally

Pet (bathing, deodorise & antiseptic) Toilet (clean, deodorise & pest control) Indoor (purify air, deodorise & pest control) 500-1000 times Closet, clothes (deodorise & antiseptic)

Upon bathing or brushing

Spray when wiping

Spray frequently

Slightly moist

Spray occasionally

1000 times

Seeding and planting (fertiliser)

Appropriate amount

Rinse once

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REFERENCES / ACKNOWLEDGEMENT

1. Sekolah Lestari, Anugerah Alam Sekitar, Institute For Environment And Development, Putrajaya, Kuala Lumpur. 2. Era Hijau, Pemantauan Pencemaran, Keluaran No.3 2009, Jabatan Alam Sekitar, Putrajaya, Kuala Lumpur. M/s: 12-13 3. Estidotmy , Kepentingan Sains dan Piawaian , Edisi 85, 25 Mac 2009, Ministry of Science, Technology and Innovation (MOSTI). M/s:24 4. Estidotmy , Agroteknologi, Edisi 78, 27 Ogos 2008, MOSTI. 5. www.fao.com 6. http://en.wikipedia.org 7. http://www.instructables.com 8. http://www.slideboom.com 9. www.o3enzyme.com 10. www.waystosaveenergy.net 11. www.slideshare.net 12. veg4planet.blogspot.com

DON T BLOW IT GOOD PLANETS ARE HARD TO FIND!!

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