2012(15A-14)-Sri K-FP.pdf

90 | PART A. NATURAL AND APPLIED SCIENCES

INTERNATIONAL JOURNAL of ACADEMIC RESEARCH Vol. 4. No. 1. January, 2012

AN INNOVATIVE NATURAL DIETARY FIBRE (NATA) PRODUCT MADE FROM SOLID WASTE OF PINEAPPLE CONCENTRATE

INDUSTRY USING ACETOBACTER XYLINUM

Sri Kumalaningsih1*, Wignyanto2, Y.S. Hendrix2, A.D. Wardanu2

1Department of Industrial Agricultural Technology, Faculty of Agricultural, University of Brawijaya, Malang, 2Postgraduate Study Program of Department of Industrial Agricultural Technology,

Faculty of Agricultural, University of Brawijaya, Malang (INDONESIA) *Corresponding author: [email protected]

ABSTRACT A large amount of solid waste does occur during the course of pineapple concentrate industry. Processing

of this solid waste for the production of nata using A. xylinum is considered beneficial not only reducing the extend of pollution problem but also producing a natural dietary fiber. Two stages of experiments were carried out. The first stage aimed to find out the best pH for each type of solid waste. A nested factorial design with two factors was used. The first factor is the type of waste (skin, pulp, and mixed skin and pulp) and the second factor is the pH (4,0; 4,5; 5,0). The second stage of experiment aimed to find out the combination treatment between initial sucrose concentrated (0,5%; 7,5%; and 10 %) and the ammonium sulphate concentration (0,3%; 0,5%; and 0,7%) for the production of Nata de Pina. Feasibility study of a small Nata plant design which could be implemented in small scale industry is carried out. The best substrate is the mixture of skin and pulp and the initial pH is 4.5. The yield of the product 53%, the thickness is 5 mm, the weight of nata is 120 g (wet basis). Further study indicated that the best combination treatment is the addition of sucrose 5% and ammonium sulphate 0.7% the thickness is 5,67mm, the weight is 217 g. Increasing the height of substrate by 50 % increased the thickness and the wet weight as well as the yield. The feasibility study was carried out based on the assumption on the production capacity of 425 kg/cicle or 5728 cup/cycle. The operational cost is $ 3,259 with the capital investment is $ 21,963.59. The selling price is $ 0,134; pay back period is 2 years one month.

Key words: nata, pinneapple waste, Accetobacter xylinum

1. INTRODUCTION

In Indonesia, pineapple (Ananas comusus (L) Merr.) is one of the most important horticultural crops which gained consumer acceptance. According to Central Bureau of Statistic [1], pineapple production in East Java in 2004 reached 444.507 tons. About 70% of the total productions is consumed as fresh product and 30% is process-sed into various food products such as jam, jelly, beverage and concentrate. Which produced a large amount of solid waste such as skin, core etc. [2].

At Pontianak, West Kalimantan Province, pineapple is processed into concentrate puree with total produc-tion capacity of 30 tons/hour which producing about 15 19,5 tons solid waste consisting of skin and also pulp [3] This solid waste has not been utilized for the production of high economical value product but is used as animal feed of low price. The use of this solid waste as raw material for the production of nata de pina by using Acetobac-ter xylinum is considered beneficial not only reducing pollution problem but also producing dietary fiber food.

Nata de Pina is microbial cellulose which become as a new type of biopolymer. Previously nata is produced from coconut water and consumed as dietary food. The manipulation of intrinsic factors such as pH, sucrose, and nitrogen compound may affect the yield of nata [4]. Several bacteria such as Acetobacter aceti, Sarcina, Agrobacterium, Rhizobium, and Acetobacter have shown the ability to synthesize cellulose from waste material. However Acetobacter xylinum has the best ability to produce cellulose [5]. Further study indicated that Accetobac-ter xylinum is an aerobic soil that ferments carbohydrate to vinegar. This organism synthesize and extrude fibril or cellulose to form a mat yielding edible product which so called nata. Fermentation production and application for the production of nata has been documented by Chwla, et all [6], and concluded that several factors such as the type of substrate and other intrinsic factor should be searched, prior to implement at industrial scale. The purpose of this study was to find out the best initial pH for each type of solid waste and the nutrient such as initial sucrose and ammonium sulphate concentration for the production of high yield of nata de pina and a feasibility study of the nata plant design profitable carried out at small scale industry.

2. MATERIALS AND METHODS

2.1. Microorganism and culture conditions Strain of A. xylinum was obtained from Agricultural Technology Laboratory, Tanjung pura, West Kalimantan

Province. The organisms were grown and maintained on nutrient agar and stored at 5 oC at Agricultural Techno-logy, division Brawijaya University East Java. A. xylinum cells were grown in Nutrient broth for 24 hours at 30 oC, harvested by centrifugation. The sediment cells were washed three times in a 0.1 % peptone solution, suspended to the desired concentration and used immediately in the experiments.

Baku, Azerbaijan| 91


Raw Materials Pineapple fruit of giant variety and solid waste consisting of skin and pulp were obtained from PT. Sari Bumi

Agrokhatulistiwa West Kalimantan province, Indonesia. Substrate Preparation Solid waste consisting of skin and pulp was seperated the skin and pulp, weight and added with mineral

water in a ratio 1:1, pressed and extracted. Starter Preparation The cels of A.cylinum were grown in 660 ml medium containing the best substrate incubated at room

temperature (25-30 C) for 7 days. Extracted Solid Pineapple Waste Preparation:

1. Laboratory Scale Experiment Preparation media from pineapple skin The pineapple fruit was peeled. The skin was collected and chopped, then blended and added with mineral

water in a ratio of 1: 1 (v/v) Preparation of mixed substrate The skin and the solid waste containing of pulp were blended in a proportion of 1: 1 (w/w) and added with

mineral water, extracted and used as substrate

2. Experimental design

First stage experiment A nested factorial design was used. The type of substrate i.e extract of skin, extract of mixed skin and pulp

and pulp as the main factor and the second factor was pH 4,0; 4,5; 5,0. The pH was nested on the substrate. The solid waste consisting of skin and pulp was mixed mineral water in a ratio of 1:1 pressed and extracted.

The extracted juice was then boiled for 10 minutes, cooled and placed in container (21 x 27 x 8 cm) and then covered with clean paper. Each container was filled with 250 ml extracted juice. The temperature of the extracted juice was controlled around 30 35 oC and the pH value was adjusted into pH 4,0; 4,5; 5,0 with the addition of acetic acid.

Second stage of experiment The best treatment obtained from the first experiment was used as substrate for the second stage

experiment. The extract was added with sucrose at three level concentration (5%; 7,5%; 10%) and ammonium sulphate at three level ( 0,3%; 0,5%; 0,7%). The best result obtained from the first and second stage of experiment was then being used for making nata de pina plant design which could be implemented at small scale industry.

3. Scale up experiment

The idea of scaling up is to increase the size of volume and amount of plate used in the process. The volume of substrate is increased to 10 times, from 200 ml to 2,0 liter / plate and the amount of plate is 9 times. Total production capacity is 10 kg of solid waste and the volume of extracted substrate become 18 litres. The best treatment obtained from the first and second stage of experiment i.e mixed solid waste extract, added with 5% sucrose; 0,7% ammonium salt and the pH is adjusted to pH 4,5; placed in plastic container incubated at room temperature ( 30 oC).

4. Feasibility Study

The Production of Nata de Pina at small scale industry. Production Capacity The processing of Nata de Pina is designed into two groups. The first group is the production of whole Nata

and the Second group is the processing of whole Nata into finished product. Raw Material Handling. The Raw Material was obtained from the concentrate of pineapple industry. After

being sortated, the refined waste was weighed and diluted with water in a ratio of 1: 1 Extraction. About 500 kg of waste was then extracted using hydraulic extractor, the extracted juice was

boiled at 1000 C for two ( 2 ) hours and then added with 5% sucrose, and 0,7% Ammoniun sulphate and agitated to homogenize all the ingredient, and then cooled and the pH is adjusted to pH 4,5 with acetic acid.

Fermentation. The extracted solid waste was then inoculated with starter of Acetobacter xylinum containing 2,0x107 cell/ ml placed in plastic container (31x23x3cm size) covered with clean paper, incubated at room temperature ( 27 300 C ) for 10 (ten) days.

Harvesting. The Nata de Pina produced was harvested and soaked for 24 hours in clean water to eliminate the sour taste, and then cut into small pieces for further processing to make beverage drink.

3. RESULTS

First stage of experiment Chemicals Characteristic of The Extracted Pineapple Solid Waste. The Analytical results presented in Table 1 shows that the substrate of Extracted solid waste made from

mixture of skin and pulp contains high crude fiber 15,43%, and valuable compounds.



Table 1. The chemicals composition of the extract pineapple solid waste

Composition Value Crude Fibre 15,43 %

Glucose 3,65 % Citric Acid 0,61 % Sucrose 8,87 % Mineral 0,115 %

Vitamin A 29,0 ( SI ) Vitamin B 0,08 mg/ 100gr

The use of extracted mixed skin and solid waste pulp was providing a promising substrate, by the

adjustment the media at pH 4.5 producing the highest thickness, wet weight and the yield of nata as shown in Table 2.

Table 2. Effect of type of solid waste and pH on the thickness, weight, and yield of nata de pina

Type of solid waste pH Thickness (mm) Wet Weight (g) Yield (%) Skin 4,0 0,49 10,00 2,86

4,5 1,97 51,00 14,57 5,0 1,13 29,33 8,38

Skin & pulp 4,0 1,26 35,00 10,00 4,5 2,37 84,67 24,19 5,0 1,00 30,33 8,67

Pulp 4,0 1,35 28,33 8,10 4,5 2,31 61,67 17,62 5,0 1,15 24,33 6,95

The formation of nata during incubation is shown in Figure 1. The microbial cellulose formed in the mixed

substrate is higher than that of the substrate which containing waste of extracted pulp only. Figure 1 showed that after 10 days of incubation the thickness of Nata was about 4.2 mm in media

containing mixed substrate.

Fig. 1. The Formation of Nata during incubation

Second stage experiment The microbial cellulose produced was proportionate to fermentation time. The organisms start to synthesize

the cellulose after 2 (two days) of incubation. Effect of sucrose and ammonium sulphate concentration on the thickness of Nata was described as in Table 3 below.. Statistical analysis indicated that there is no significant interaction between treatments. The results are given in Table 3.

Table 3. Effect of sucrose and ammonium sulphate concentration on the thickness of Nata

Sucrose (%) Ammonium Sulphate (%) 0,3 0,5 0,7

Thickness 5 4,67 4,9 5,67 7,5 3,50 3,73 3,67 10 1,83 2,17 2,17

Table 3 showed that sucrose and ammonium salt concentration has affected individually. The best sucrose

concentration is 5%. Increasing the sucrose concentration caused the media become viscous and inhibited the organisms to grow. Further study was to improve the thickness weight and yield of nata, by increasing the height of substrate as shown in Table 4.

0

1

2

3

4

5

2 4 6 8 10

Pulp

Mixed Substrate

Days

Thickeness(mm)

Baku, Azerbaijan| 93


Table 4. The Effect of height of substrate on the thickness wet, weight and yield of Nata

Height of Media ( cm )

Thickness ( mm )

Weight ( g )

Yield ( % )

1,5 5 190 54,29 3,0 15 270 98,5 4,5 16 275 99,9

The height of media also affected the production of Nata, the maximum height is 3 cm of the reactor vessel.

Presumably the bigger the amount of substrate resulted in the more valuable compound and dissolved oxygen which support the extended of cellulose production. Increasing the content of volume of media has no significant effect. These results were then used for scale up experiment.

Fiber content The sucrose concentration affected the fibre content but the addition of ammonium sulphate has no

significant effect. Table 5 showed that there is no significant interaction between sucrose and ammonium salt concentration on the fibre content of nata de pina. Table 5 shows the fiber content of nata after various treatment and times.

Table 5. Effect of sucrose and ammonium sulphate concentration on the fiber content of nata (% ).

Sucrose (%) Ammonium Sulphate (%) 0,3 0,5 0,7 Total

5 2,77 3,05 3,55 3,12 7,5 2,37 2,27 2,47 2,37 10 1,35 1,69 2,29 1,78

The highest amount of fiber content is obtained in treatment containing 5% sucrose and 0,7% Ammonium

Sulphate, About 3,12% (w/w) of fiber is achieved.

Scale Up Experiment Result obtained from the scale up experimentalism indicated that increase volume of extracted media from

200 ml to 2 liter or total volume of media is 18 liter. Result is an increase in the yield of Nata. This may be due to the height of the media that used is larger than media at laboratory scale.

From the scale up experiment the Nata de Pina produced is 10.6 kg and the thickness is 7 11 mm, the weight is 487.560 gram, and the yield is 64% which higher than that obtained from the laboratory experiment which is only 54%.

Financial Analysis Process of production. The production of nata de pina is designed as a continuous process which divided into two part of

processing. The first is the production of nata de pina at large scale prior to package and the second part is the production of nata in plastic cups containing 100 ml of nata/cup. Production capacity is 462.51 kg nata/cycle or 5728 cups/day, or the actual capacity is 425 kg/ cycle.

Capital Investment Total = USD 21,722 Operational Cost = USD 3,259 Fixed Capital = USD 18,464

Feasibility indicators Feasibility indicators of nata de pina industry indicated by the value of IRR is 56.80%, NPV of USD 64,382;

3.7 profitability index, payback period of 2 years 1 month; break even point at the production level is 33.39% and net B / C 1.39. These results show this project is feasible.

4. DISCUSSION

As stated by Anastasia and Afrianto [7] pineapple fruit of giant variety used in this experiment has thick skin, about 15 20% of fruit consisting of skin, which still contained a large amount of fiber, sucrose, glucose, mineral and vitamin, where as the pulp as waste containing organic acid. These entire valuable compound may support the synthesized of cellulose.

Apparently the use of mixed skin and pulp at pH 4.5 cause the media has pronounced and environmental condition which favored the organism to grow. The formation of acid during fermentation resulted in lowering the pH level and hence inhibited the wild organism to grow. It is therefore this treatment has the highest thickness of and obviously influence the wet weight and the yield after 10 days of incubation (the yield is in the range of 2, 66 24, 19 %).

The crude fiber presence in the media has the potential role to distribute the nutrition during fermentation. On the other hand crude fiber contains various micro elements such as Fe, Mn, Mg which may support the activity of enzyme [8].



This evident indicated that the substrate has high nutrition value, although there are no micro particles as stated by Chng and Muhammad [9] (Figure.l). The Addition of sucrose is very important to sustain the formation of Nata (Table 3). Presumably sucrose concentration up to 5 % could stabilize the existence of carbon source thus the microbial cellulose synthesize could be sustained, as stated by Mashudi [10].

Nata de pina produced in this study is higher than that nata de coco, which only contain 2,5% fiber..Apparently the bigger or larger the fermentation container increases the amount of dissolved oxygen, As stated by Tantration et.al. [11], the dissolved oxygen presence in the media may increase the glucomic acid of the media which support the organism to produce Nata.

During fermentation the extrinsic and intrinsic factor play an important role. According Mashudi [10], the medium used to grow microorganisms must be able to transform and meet the needs of carbon compounds and energy, as an element of macro-nutrients such as P, K and Mg, micronutrients and growth factors such as vitamins and amino acids. Minerals and vitamins needed in the media establishment as micro-elements of nata materials required but in small amounts. Minerals and vitamins will be used by the bacteria in it is life. Besides, the medium components that must comply with the requirements of microbes, before being used should be in a state of sterile medium, not overgrown by other microorganisms that are not desired.

5. CONCLUSION

Skin waste can be used as growth media for the production of nata de pina. The best substrate is the mixture of skins and pulp and the initial pH 4.5. The yield of the product 64%, the thickness is 12 mm. Analysis result of the industry shows this project is feasible.

REFERENCES 1. Biro Pusat Statistik, 2007, Survei Pertanian Produksi Buah-buahan di Jawa, BPS, Jakarta-Indonesia 2. Tahir, Sri Sumarsih and Shinta Dwi Astuti. 2008. Kajian penggunaan limbah buah nenas local

(ananas comosus, l) sebagai bahan baku pembuatan nata. Makalah Seminar Nasional Kimia XVIII, Jurusan Kimia FMIPA UGM Yogyakarta

3. Sianipar J. Krisnansimanihuruk K. dan Batu Bara L.P. (2006). Evaluasi Tiga Jenis Limbah Pertanian, Sebagai Pakan Kambing Potong. Seminar Nasional Teknologi Peternakan dan Veteriner Medan

4. Misgiyarta, 2011. Teknologi Pembuatan Nata de Coco, Balai Besar Penelitian dan Pengembangan Pascapanen Pertanian. http://pascapanen.litbang.deptan.go.id/media/berita/misgiyarta-natadeCoco. pdf. Acces date: March 10th 2011

5. Tsuchida T. and Yoshinaga F. 1997. Production of Bacterial Cellulose by Agitation Culture System. Pure & Appl, Chem., Vol.69, No. 11, pp.2453-2458

6. Chawla P.R. Ishwar B.B., Shrikant A. S and R.S. Sighhal. 2009. Microbial Cellulose: Fermentative Production and Applications. Journal of Food Technology, Biotechnology. India

7. Anastasia and Afrianto.2008. Nata de Sewed Quality in Different Citrus Addification. Proceeding of National Science and Technology II 2008. University of Lampung. Lampung

8. Misgiyarta, 2011. Teknologi Pembuatan Nata de Coco, Balai Besar Penelitian dan Pengembangan Pascapanen Pertanian. http://pascapanen.litbang.deptan.go.id/media/berita/misgiyarta-natade Coco. pdf. Acces date: March 10th 2011

9. Chng and Muhammad.2003. Evaluation and Optimization of Microbial Cellulose (nata) Production Using Pineapple Waste as Substrate. Chemical Engineering. UTM. Malaysia.

10. Mashudi. 1993. Mempelajari pengaruh penambahan sumber nitrogen dengan berbagai konsentrasi pada pembuatan nata de coco. Skripsi jurusan teknologi pangan dan gizi, Fateta, IPB.Bogor

11. Tantration S., Tammarate W. Krusong, Bhattarakosol, Phunsri. 2005. Effect of dissolved oxygen on cellulose production by Accetobacter sp. Journal Sci. Res. Chula Univ. 30. Pp. 179-186

Documents

2012(15A-14)-Sri K-FP.pdf