UNIVERSITY OF SAN CARLOS Nasipit, Talamban, Cebu City

ChE 313 L Industrial Chemistry Laboratory GROUP1- WEDNESDAY 7:30 10:30 am

Production of Nata de Coco

LABORATORY INSTRUCTOR: Dr. Andresa S. Allera GROUP 1 MEMBERS: Bote, Mae Bernadette Doroy, Clarince Joyce Dividina, Rosauro Lasala, Jovan Andrew

I.

Objectives:

a) Apply the fundamental principles of aerobic fermentation by bacteria in food production b) Describe the chemical reactions associated with nata production c) Determine possible economically viable sources of substrate for making nata d) Qualitatively describe the effect of substrate and growth conditions of bacteria on nata production e) Identify the different applications of nata f) Prepare 500 grams of nata appropriately packed for presentation and submission II. Historical Background

Being an archipelago, the Philippines home many species of coconut. That's why Filipinos utilize coconut trees, from its roots to its leaves, and even the newly formed buds or soot. One of the main use of coconut tree is its fruit, the coconut, which is composed mainly of lipids. Microorganisms ability to synthesize new products from one substrate had been employed since ancient times. Using the process of fermentation, its products which includes wine, vinegar and bread had been the major source of nutritious foods for us. Using this process, Filipinos were able to create a native delicacy using coconut cream. Nata de Coco is the name best known in the Philippines to describe a chewy, translucent, jelly-like food product processed from coconut water with the help of bacteria, Acetobacter xylinum. Nata de Coco was introduced in Indonesia around 1987. Nata de coco is Spanish for "cream of coconut". Cream in this sense means the fat from the coconut milk. The Spanish name is a result of Spain's colonization of the Philippines. Nata is actually cellulose mat produced by certain strains of Acetobacter xylinum through traditional static fermentation. Although this bacterial cellulose has been subjected for extensive investigation, especially on the fiber microstructure, biosynthetic pathway, and production of cellulose in agitated culture; very few studies was conducted to elucidate the biochemical and microbiological aspects of fermentation in nata production for food. Nata De Coco was embraced by the public as an affordable desert. Gaining mass popularity because of it's taste the notoriety spread nationwide quickly. In the early 90s, Nata De Coco had a surprising boom in the export market when it created a surge of demand from Japan. Armed with simplified method of production, the Phil. Gov't. through it's livelihood program administered via DOST (Department of Science and Technology) implemented the transfer of technological information and production training workshop among livelihood projects and small scale private enterprise to keep up with the demand. The recorded revenue in 1991 for export in Japan was around $ 170, 000.00. In the late 90's the export demand simmered down as other countries like Thailand, Indonesia and Malaysia join bandwagon of production and export trade.

III.

Theoretical Background

Nata de coco is produced from coconut cream and some bacterial action,through fermentation. Basically it's the croorganisms who works to come with the final product. First, a mature coconut should be obtained and cut into two exposing its meat. The meat is grinded and pressed releasing the co called coconut cream. A little water can be added for better extraction. Upon extracting the coconut cream, set it aside first. Then a starter medium containing the Acetobacter species, a type of acid forming bacteria, is added to the coconut cream prepared earlier. There are two ways on doing this, the first is on a sterile environment, avoiding contamination, and second is non-sterile. Upon mixing, the resulting mixture is transferred into a tray and covered by a cheesecloth. (If you want to disinfect the tray, rinse it with a boiling water twice.) Place the tray with the nata de coco mixture into an incubator and leave it there for about a week under room conditions. Make sure that any movement of the tray will be avoided for the said period. A cellulosic layer is expected to be formed within a week. Basically that is the nata de coco already. It should be harvested immediately and stored in a container with water. In a general sense, fermentation is the conversion of a carbohydrate such as sugar into an acid or an alcohol. More specifically, fermentation can refer to the use of yeast to change sugar into alcohol or the use of bacteria to create lactic acid in certain foods. Fermentation occurs naturally in many different foods given the right conditions, and humans have intentionally made use of it for many thousands of years. Acetobacter xylinum is a gram-negative, aerobic bacterium that has long served as a model organism for the study of bacterial cellulose synthesis, primarily because of the large quantities it produces. A single A. xylinum cell is capable of polymerizing 200 000 glucose molecules per second into -1,4-glucan chains which are then excreted into the surrounding medium forming ribbon-like bundles of microfibrils. The produces crystalline fibres resemble in width and structure average fibrils form many plants and algae. The fibres are formed in the membrane by cellulase synthase and consequently secreted from a row of 50 80 pore-like synthetic sites along the longitudinal axis of the cell. The formation of this floating cellulose matrix is thought to allow A. xylinum, an obligate aerobe, to grow in the higher oxygen tension at the surface of the medium. Some practical applications of bacterial cellulose include: sensitive diaphragms for stereo headphones, thickeners for foods, paint, ink and adhesives. It has also been used as short term protection for burned skin prior to skin grafting. Most commercial applications of cellulose produced by Acetobacter have employed static batch culture to produce sheets

of cellulose that can be sterilized to kill the bacteria and washed to remove microbial waste product contaminants. One large scale commercial application is the production of bacterial cellulose in a nonaseptic medium of sugar-enriched coconut milk in the Philippines, yielding an edible product called nata de coco. This food product has become popular in Japan as a high-fiber, low-calorie food. Recently, Japanese investigators have developed a technique to harvest Acetobacter cellulose from shallow culture pans with a windup roller device to pull the cellulose slowly from the surface of the culture medium. High yield commercial production of bacterial cellulose will require agitated (shaking) culture techniques. There has been some limited success in the laboratory studies producing cellulose in this way, but, at present, there are no commercial applications of this process.

IV.

Materials/ Apparatus/ Equipment Used 1 kg. fresh coconut, grated 325 mL or 1 cups glacial acetic acid 600 g or 3 cups refined sugar 500 ml or 2 cups coconut water 12 L tap water 2 L nata starter Cheese cloth Strainer Large mixing bowl (1) 1L beaker (1) 400mL beaker Fermenting basins/jars

V.

Definition of Terms 1. Fermentation - The chemical breakdown of a substance by bacteria, yeasts, or other microorganisms, typically involving effervescence and the giving off of heat. 2. Microfibril - A small fibril in the cytoplasm or wall of a cell, visible only under an electron microscope, and typically aggregated into coarser fibrils or structures. 3. Cellulose - An insoluble substance that is the main constituent of plant cell walls and of vegetable fibers such as cotton. It is a polysaccharide consisting of chains of glucose monomers.

VI.

Processes/ Reactions Involved Acetobacter xylinum is a gram-negative, aerobic bacterium that has long served as a model organism for the study of bacterial cellulose synthesis, primarily because of the large quantities it produces. A single A. xylinum cell is capable of polymerizing 200 000 glucose molecules per second into -1,4-glucan chains which are then excreted into the surrounding medium forming ribbon-like bundles of microfibrils.

The produces crystalline fibres resemble in width and structure average fibrils form many plants and algae. The fibres are formed in the membrane by cellulase synthase and consequently secreted from a row of 50 80 pore-like synthetic sites along the longitudinal axis of the cell. The formation of this floating cellulose matrix is thought to allow A. xylinum, an obligate aerobe, to grow in the higher oxygen tension at the surface of the medium.

Bacterial reaction C2H5OH + O2 Alcohol CH3COOH + acetobacter acetic acid H2O water

In this type of fermentation, starter cultures (similar to those used in the dairy industry) are recommended. Not only do starter cultures ensure consistency between batches, they speed up the fermentation process as there is no time lag while the relevant microflora colonize the sample. Because the starter cultures used are acidic, they also inhibit the undesirable micro-organisms.

VII.

Product Characteristics/ Applications

Nata de coco is most commonly sweetened as a candy or dessert, and can accompany many things including pickles, drinks, ice cream, puddings and fruit mixes. Nata de coco is highly regarded for its high dietary fibre, and its low fat and cholesterol content. Strips of nata de coco are used in mass-produced bubble tea drinks as a healthier alternative to tapioca. It is mixed in one of the most popular refreshment, the halo-halo, with other stuffs like red beans, macapuno, sweetened bananas and sweet potatoes, ube and leche flan.

VIII.

Procedure ENGLISH VERSION 1. Mix grated coconut with water. Strain thru cheesecloth. 2. Add the rest of the ingredients and mix. 3. Pour the mixture into clean sterile fermenting basins or jars (2 inches depth). Set aside starter for use in 3-5 days. 4. Ferment for 12-14 days. 5. Harvest. Wash.

BISAYA VERSION
1. 2. 3.

Isagol ang kinagud na lubi sa tubig. Isala ang tuno gamit sa cheesecloth. Idungag ang ubang ug isagol. Ibubo ang mga sinagol sa mga limpyo ug sterilized na butanganan. Pasagda-i ang starter para gamiton ing 3-5 ka adlaw.

4. 5.

Ipa-ferment sa sulod sa 12-14 ka adlwa. Anihon. Hugasi.

IX.

Schematic Diagram

X.

Materials Costing

Materials Coconut milk Nata starter 325 ml Glacial acetic acid Cheese cloth Sugar XI. References

Cost Php 25/ whole coconut Php 100 .00/L Php 22.75 Php 32.00 Php 35.00/kilo

http://www.agriqua.doae.go.th/worldfermentedfood/I_7_Suwanto.pdf http://muhtaufiqmunawar.blogspot.com/2009/02/pohon-kelapa-termasuk-dalamkeluarga.html http://yhep.yolasite.com/bacterial-cellulose.php http://guides.wikinut.com/Fermented-Food%3A-Nata-de-Coco/3f48k4jg/ http://guides.wikinut.com/Fermented-Food%3A-Nata-de-Coco/3f48k4jg/ http://www.nabt.org/websites/institution/File/pdfs/american_biology_teacher/2000/0 62-06-0442.pdf http://www.fao.org/docrep/x0560e/x0560e10.htm