Bioenzymes: Effectiveness in Waste Reduction System

CHAPTER 1BACKGROUND

INTRODUCTIONThe outset of globalization, technological and industrial development eventually resulted to an increased solid waste generation worldwide. Developed countries in Europe and the United States of America are now conducting studies aimed at providing solutions to their waste management and disposal problems. (Some countries even exported tons of garbage in container vans in developing countries like the Philippines) In Cabanatuan City, considered as the economic heart of Nueva Ecija and is the focus of a metropolitan area of more than 300,000 people, wastes disposal is one huge problem. Cabanatuan City residents, industrial and business establishments generated tons of garbage every day. 50% of which is biodegradable 35% recyclable and 15% residual. To address the growing problem of waste disposal, the City Government of Cabanatuan instituted ways of improving its existing dump sites and transfer facility stations.The Valle Cruz Transfer Facility Station in Cabanatuan was temporarily closed to give way for its rehabilitation. The rehabilitation of the said facility was aimed at removing odorous pollution that is affecting the residents within and around the vicinity. These pollutants are mostly organic solid wastes which has the highest percentage amount based from the previous Waste Analysis Characterization System (WACS), 2009. These solid wastes are non-soluble materials that were discarded in a solid or semi-solid form. The semi-solid form of waste has the quality of both solid and liquid- like sludge.In Accordance with Ra 9003 Rule X, Section 2, Transfer stations shall be designed and operated for efficient waste handling capacity and in compliance with environmental standards and guidelines set in pursuant to the Act. Provisions for vector, odor, litter and dust control shall be included (Ra 9003 Rule X, Section 2 Criteria a). In order to comply with this criterion, the use of enzymatic solution is the most viable option.Enzymatic solutions are natural, nontoxic substance that uses enzymes which helps the bacteria to digest organic materials directly. Enzymatic solutions are formulated specifically to dispose of soils safely, economically and rapidly. They contain the necessary quality and quantity to specific enzyme systems, both aerobic and anaerobic bacteria, and microbial nutrient. Enzymatic solution works quickly and efficiently in the digestion of chemical and organic waste with no odor or noxious gas emissions. These microorganism and right type of enzymes break the waste apart and the bacteria itself converts this waste into two basic compounds: carbon dioxide and water. This resulted in faster degradation of wastes with minimal residual component, and the absence of foul odors. Geographically, Valle Cruz is located in a residential and agricultural area where emission of odorous pollutants produces grave effects on the health of the constituents. Extension of the said facility to accommodate extra tons of wastes costs large amount of taxpayers money and land conversion causes social and economic nuances. A more viable solution is the use of alternative means of waste reduction and anti-pollution technology, that is, the application of enzymatic solutions. Thus, it is therefore necessary to conduct this study entitled Comparative analysis of some Enzymatic Solutions: Its Effectiveness to waste reduction system to determine the effectiveness of the application of some enzymatic solution in reducing environmental wastes and pollution caused by organic solid waste. Furthermore, this study can help not only in degradation and deodorization of solid waste but also in providing a clean and health conducive environment.OBJECTIVES OF THE STUDY1. To compare H2O (negative control), commercial enzymatic solution (positive control) and non-commercial enzymatic solution (experimental) in terms of organic waste degradation and odor.2. To estimate the total time that can be saved using experimental and control group in the degradation of organic solid wastes;3. To determine the impact of enzymatic solutions in environmental conservation.HYPOTHESIS The study was tested for the following hypothesis. The enzymatic solution boosts the degradation of organic solid waste and reduced the odor. Not all enzymatic solutions have the same strength, some can only reduce the odor and some can boost the degradation and others can do both. There is a significant change in the size, shape, and weight of the organic wastes applied with enzymatic solution.

SIGNIFICANCE OF THE STUDYThe study will be beneficial to the following:Students: the result of this study will serve as references for further studies on subjects dealing with the environment and its conservation.Local Government Unit: The result of this study is very beneficial to LGUs. The problem in local sanitation as well as pollution caused by the burning of wastes is readily solved.Residents: The elimination of disease causing odorous wastes is the greatest relief that residents within the vicinity can be experience.Environment: The study aborts the necessity of further extension of the dumpsite therefore land conversion which is of course very harmful to the ecosystem as well as the social environment can now be avoided.Transfer facility workers: the huge reduction of waste residue and the elimination of foul odors will reduce the burden on the workers in the said facility. Efforts, time and manpower will now be saved as physical handling of the waste materials is now minimal.Future Researcher: This study will serve as their stepping stone in furthering their research on the effectiveness of the said application and will serve as reference for future testing, experimentation and evaluation of its effectiveness.

SCOPE AND DELIMITATIONThe focus of the study is to determine the effectiveness of commercial and non-commercial enzymatic solutions in deodorizing and decomposing organic waste to determine its usefulness in solid waste reduction system. This study did not focus on the different components of enzymatic solutions. Enzymatic test was not conducted but was recommended for the continuation of further study by other researchers, because Bachelor of Science in Environmental Science focuses only on social environmental problems. Micro aspects such as enzyme activity and other microbiological parameters are not under the description of the course.The Experimental procedure was done at the College of Arts and Sciences (CAS) Laboratory, NEUST, Cabanatuan City.

Conceptual FrameworkTreatment Effect Impact Odor reduction of organic waste

Enhanced degradation of organic waste

Saved time.

T1Negative controlApply pure water

T2Positive controlApply commercial enzymatic solution on the sample

ExperimentalT3Apply non-commercial enzymatic solution on the sample

T1Negative Control Does not reduce bad odor and have a slow degradationT2Positive control Reduce bad odor after 2 to 5 weeks. Decomposed after 14 to 30 daysT3Experimental Reduce bad odor after 2 to 5 weeks but not as effective as T2

Definition of termsFor clear understanding of the terms used in this study definitions of unfamiliar words are listed alphabetically.Aerobic bacteria- aerobe is an organism that can survive and grow in an oxygenated environment.Anaerobic bacteria- are anyorganismthat does not requireoxygenfor growth. It may react negatively or even die if oxygen is present.Alpha amylase-a proteinenzymeEC3.2.1.1thathydrolysesalpha bonds of large, alpha-linkedpolysaccharides, such asstarchandglycogen, yielding glucoseandmaltose.It is the major form ofamylasefound in humans and other mammals.It is also present in seeds containing starch as a food reserve, and is secreted by many fungi.Biocatalysis- is the use of naturalcatalysts, such as proteinenzymes, to perform chemical transformations onorganic compounds. Both enzymes that have been more or less isolated and enzymes still residing inside living cells are employed for this task.Biodegradable- materials that is capable of being broken down especially into innocuous products by the action of living things (as microorganisms).Bioenzymes/Bioenzymatic cleaner- Anenzymatic cleaneris a biodegradable, non-toxiccleaningagent.Decomposition(or rotting) - The process by which organic substances are broken down into simpler forms of matter.Deodorization- is the process of removing foul odor of a substance.Enzymes- Enzymes are macromolecular biological catalysts. They are responsible for thousands of metabolic processes that sustain life.Glucanases- areenzymesthat break down aglucan, a polysaccharide made of severalglucosesub-units. As they performhydrolysisof the glucosidic bond, they arehydrolases.used in enological practices during the aging process of wine, particularly when aged on lees with microxygenation. The enzyme aids in autolysis of yeast cells to release polysaccharides and mannoproteins, which is believed to aid in the color and texture of the wine.Odor- Anodoror odour or fragrance is caused by one or more volatilized chemical compounds, generally at a very low concentration, that humans or other animals.Pollution- the action ofpollutingespecially by environmental contamination with man-made waste;also: the condition of beingpolluted.Recyclables- are materials that can be recycled and use for other uses.Rehabilitate- to restore to a former state (as of efficiency, good management,Residuals- Materials that cannot be processed to be useful again.Respiromentry- a general term that encompasses a number of techniques for obtaining estimates of the rates of metabolismofvertebrates,invertebrates,plants, tissues, cells, ormicroorganismsvia an indirect measure of heat production (calorimetry).WACS- Waste Analysis characterization system waste Transfer station- a building or processing site for the temporary deposition ofwaste.

Chapter IIREVIEW OF RELATED LITERATURERelated literatureThis chapter presents a review of literatures and studies which are relevant to the present study.Biological Information of Commercial Enzymatic SolutionsThis brand is made up of blend of different types of live, highly selected beneficial microbes (bacteria, yeast, actinomytecete, and molds), Enzymes, vitamins and organic acids mixed with specially prepared nutrients in solutions.Through the process of advanced fermentation technology, the microbes in the product where trained to grow harmoniously without antagonizing each other. This is based on several years of research and development work. The microbes have the capacity to produce powerful enzymes which could digest or degrade animal and plant tissues, proteins, fats and cellulose. The combined actions of this biological fermentation are responsible for the effective deodorizing, sanitizing, and decomposing properties of this commercial enzymatic solution.According to the test report of the National Institute of Molecular and Biotechnology (Biotech) they have determined the enzymatic activities of alpha amylase, beta glucanase, cellulose, pectinase, and alkaline in the liquid solution of biozyme.According to the textbook Regulation and Control of Metabolism in Bacteria As a form of Catabolite Repression, the glucose effect serves a useful function in bacteria: it requires the cells to use the best available source of energy. For many bacteria, glucose is the most common and readily utilizable substrate for growth. Thus, it inhibits indirectly the synthesis of enzymes that metabolize poorer sources of energy.Here are some studies that might contribute to the present study. Related Studies:According to A. Schmid (2001) the use of biocatalysis for industrial synthetic chemistry is on the verge of significant growth. Biocatalytic processes can now be carried out in organic solvents as well as aqueous environments, so that a polar organic compounds as well as water-soluble compounds can be modified selectively and efficiently with enzymes and biocatalytically active cells. As the use of biocatalysis for industrial chemical synthesis becomes easier, several chemical companies have begun to increase significantly the number and sophistication of the biocatalytic processes used in their synthesis operations.Biocatalysisis the use of naturalcatalysts, such as proteinenzymes, to perform chemical transformations onorganic compounds. Both enzymes that have been more or less isolated and enzymes still residing inside living cells are employed for this task.Sited from the book Green Chemistry for Environmental Remediation by Rashmi Sanghi and Vandana Singh (2012) enzymes are biocatalysts, which speed up chemical reactions without undergoing any net chemical change during the reaction. Enzymes are known to perform a multitude of chemical reactions and are commercially exploited in the detergent, food, pharmaceutical, diagnostics and fine chemical industries. More than 300 enzymes have been exploited to date, and majorities have been isolated from mesophilic organisms which function in a narrow range of ph, temperature and ionic strength. The microorganisms from exotic environments (Extremophiles) are considered an important source of enzymes and their specific properties are expected to result in novel process applications.According to Ms. Payel Sarkar (2011).The degradation of organic wastes by the bacterial consortia is highly significant. It reduces the time span of degradation and produces no foul odor. The use of microbial consortium generated through natural selection or improvement of the performance of these microorganisms in organic kitchen waste degradation through genetic manipulation, may be the best option for the efficient treatment of organic kitchen waste or domestic wastewater in the near future. The pretreatment of food waste can be used for biological solubilization and mineralization in garbage disposal system which is a novel approach.According to the study entitled Solid Waste Management by Use of Effective Microorganisms Technology by Ms. Dipti Kumar Kale and Dr. P.D. Anthappan (2012).The quantity of waste generated due to urbanization pose a major problem of proper and adequate treatment, so that its final disposal should not cause environment pollution. Many scientific techniques have been discovered for proper waste management, followed by its disposal. But today, it is necessary to use such a method which can convert waste into reusable by-product. Thus with the aim to combat pollution, this research topic highlights the use of one of the simple and easy method; Effective Microorganism (EM) Technology, is the one in which organisms from natural sources are utilized to convert waste into a reusable byproduct. It has desirable effect of increase in organic carbon (OC), organic matter (OM), nitrogen (N), phosphate (PO4) and potassium (K) content of waste. In this study it was found that OC, OM, N, PO4, K, increased at an average of 22.67%, 39.09%, 1.84%, 95mg. /lit, 2.88mg./liter respectively when compared with control where the EM was not added. Based on the result obtained, it was concluded that the use of EM technology has desirable effect which has been observed by many studies.According to Natarajan, 1988 the amylolytic activities of alpha-amylase extracted from Mirror carp (Cyprinuscarpio) and Nile tilapia (Oreochromisniloticus) were significantly reduced by purified amylase inhibitor albumin of wheat when tested under in vitro conditions. The action of this inhibitor was rapid and maximum levels of inhibition were attained within 20 minutes. For both carp and tilapia, the enzyme residual activities after inhibition were found to be related inversely to inhibitor concentration and positively to the initial enzyme activity levels. The curvilinear relationships between these parameters were explained by deriving equations of the type: A2 = a+b A1 - c I + d I2 where a, b, c and d are constants, Ai = Initial amylase activity (mU/min), A2 = Residual amylase activity (MU/min), I= Inhibitor concentration as ug protein. Inhibitions were greatest for amylases from gut tissue and owest for amylases from gut fluids. 1ug of purified inhibitor was found to contain a potency, to reduce 298 Units of carp gut tissue alpha-amylase and 532 Units of tilapia intestinal tissue alpha-amylase, by 50%. When amylase inhibitor extracted from wheat was incorporated in the feed of carp in its active form for three weeks, it caused a significant reduction in the specific growth rate to only 0.16%/day, while in carp fed autoclaved inhibitort such reduction in growth was not seen and the SGR was maintained at over 1.00%/day. However, despite the presence of active inhibitor in the intestine, the fish were able to maintain alpha-amylase activities in the gut contents at a level similar to that in fish fed denatured inhibitor. This was achieved by hyperactivation of enzyme secretions in the tissues of hepatopancreas and intestine. Hepatopancreas from fish fed active inhibitor exhibited more than two-fold increase in amylase, activity compared to those fed denatured inhibitor. By the third week of the experiment this difference in enzyme activity levels was not apparent but there were also no indications of adaptation or improvement in growth rate. Degenerations in hepatopancreas were also not apparent. Feeding carp with diet containing wheat with its inherent content of inhibitor also caused pancreas hyperactivity and some reduction in growth rate for a short period in comparison to those fish fed autoclaved wheat. In carp, the alpha-amylase activity did not vary depending on the raw or gelatinized nature of starch, both forms elicited equal increases in enzyme activity. However, autoclaving wheat, though effective in inactivating the inhibitor, was found to lower the biological value and digestibility of wheat proteins. Contrary to the result of the carp trials in Nile tilapia, the growth was not significantly reduced by feeding on diet containing active inhibitor and a SGR of 1.57%/day was recorded in comparison to 1.81i/day in tilapia fed denatured inhibitor. Samples of stomach and intestinal contents collected 4 hours after feeding did not reveal the presence of active inhibitor. Apparently the acidic protease, pepsin, in the stomach of tilapia caused the total destruction of the inhibitor in the diet before the contents were passed into the intestinal region. The presence of active amylase inhibitor in tilapia feed did not affect the digestibilities of starch and protein in the diet. Both the groups were able to digest carbohydrates and protein to levels of over 90%. The implications of these results are discussed in relation to feed formulation and fish nutrition.According to Rungkan Klahan1, Nontawith Areechon1, Ruangvit Yoonpundh1 and AruneeEngkagul the activity of protease, amylase and lipase from the pseudo stomach, upper intestine, lower intestine and liver of 5.7, 35.8 and 92.1 g male Nile tilapia (Oreochromisniloticus L.) was determined. Protease activity had an optimal pH of 12, 9 and 9 for the 5.7, 35.8 and 92.1 g fish respectively and the highest specific activity was found in the 35.8 g fish. Amylase activity had an optimal pH of 6, 7 and 2, respectively and the highest specific activity was found in the 92.1 g fish. Lipase activity had an optimal pH of 8, 7 and 8, respectively and the highest specific activity was found in the 35.8 g fish. In all sizes of tilapia, amylase activity was significantly higher in the liver than other organs (P