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Anny Sulaswatty and. Karossi, A.T; Fermentation in Industrial Scale Country Report 1990

FERMENTATION IN INDUSTRIAL SCALE INDONESIAN COUNTRY REPORT

Anny Sulaswatty and Karossi A.T. R & D Centre for Applied Chemistry - LIPI Indonesian Institute of Sciences Jalan Cisitu Sangkuriang, Bandung, 40135, Indonesia

INTRODUCTION Biotechnology has become the focus of worldwide attention due to new and innovative developments in several fields. For centuries, microorganisms have been used in food production such as wine, beer and vinegar or in the production of solvents such as alcohol. This kind of biotechnology is considered an old one because the primary force behind the production was merely a result of selection of microorganisms. The new biotechnology, on the other hand, involves the manipulation of the interior structure of the microbes mainly through recombinant DNA technology. In a developing country where the economy is agriculturally based, such as Indonesia, biotechnology can play a significant role in the development of the agricultural sector, in the development of its agro based industries, the improved utilization of its agricultural by- product and better management of the environment. A few institute have already focused their activities on developing prototypes or production side. However, many of these data are not properly utilized yet. Consequently the industrial production or commercial scale in Indonesia is still small.

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Anny Sulaswatty and. Karossi, A.T; Fermentation in Industrial Scale Country Report 1990

This paper will give a brief account of current state of the scale up and technology transfer of biotechnological processes in Indonesia. STATUS OF FERMENTATION INDUSTRIES IN INDONESIA The major fermentation products currently produced in industrial or commercial scale in Indonesia include monosodium glutamate, glutamic acid, glucose and fructose syrup, ethanol, beer, fermented fish sauce, soybean sauce, yeast, cheese, tempe and citric acid. Production data for some of these product are presented in Table 1 (1, 2).
Table 1. Production of Indonesian Fermentation Industries

Commodity and company A. Glutamic acid * 1. PT. Ajinomoto Indonesia 2. PT. Miwon Indonesia 3. PT. Sasa Inti 4. PT. Ajinex Inti 5. PT. Palur Raya 6. PT. Bumas Rajawali Prabhawa 7. PT. Glutama Indorasa 8. PT. Cheil Samsung Astra 9. PT. Monosari Glutama 10. PT. Indomiwon Citra Inti 11. PT. Indo Miki 12. PT. Sahid Mekosin Kujang Lestari

Location of plants

Yearly capacity (ton)

Mojokerto Gresik Probolinggo Mojokerto Solo Sidoarjo Merak Pasuruan Lampung Lampung Batang, Pekalongan Krawang

36,000 45,000 44,000 27,000 12,700 30,000 24,000 20,000 18,000 18,000 10,000 10,000 294,700

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Table 1. continued
Commodity and company
B. Mono sodium glutamate * 1. PT. Miwon Indonesia 2. PT. Sasa Inti 3. PT. Ajinomoto Indonesia 4. PT. Bumas Rajawali Prabhawa 5. PT. Ajinex Inti 6. PT. Glutama Indorasa 7. PT. Cheil Samsung Astra 8. PT. Monosari Glutama 9. PT. Indomiwon Citra Inti 10 PT. Palur Raya 11. PT. Indo Miki 12. PT. Sahid Mekosin Kujang Lestari 13. PT. Rena Jaya 14. PT. Sasa Fermentasi 15. PT. Foomaco 16. PT. Indo Vetsin Gresik Probolinggo Mojokerto Sidoarjo Mojokerto Merak Pasuruan Lampung Lampung Solo Batang, Pekalongan Krawang Semarang Sidoarjo Ungaran Solo 45,000 44,100 36,000 30,000 27,000 24,000 20,000 18,000 18,000 12,000 16,800 10,000 3,600 3,600 1,800 2,200 311,100

Location of plants

Yearly capacity (ton)

C. Yeast * 1. Bakers Yeast 2. Fodder Yeast 3. Bread Improver Depok Depok Depok 1,000 1,000 20 2,020

D. Ethanol * 1. PT. Indo Acidatama 2. PT. Aneka Kimia 3. PT. Jatiroto 4. PT. Madusari Murni 5. PT. Comal 6. PT. Basis Indah 7. PT. Permata Sakti 8. PT. Palimanan 9. PT. Madu Baru 10. PT. Molindo Raya 11. PT. Starsaco 12. PT. Padaharja 13. PT. Nabati Sarana E. Citric acid * 1. 2. 3. 4. 5. 6. PT. Budi Acidjaya PT. North Aspac Chem. Ind. PT. Sari Idaman PT. Budi Alam Kencana PT. Kali Jaya King Raya PT. Kancing Mas Jaya Lampung Jakarta Surabaya Lampung Jakarta Semarang 2,730 1,080 120 1,000 3,000 1,200 9,130 Solo Mojokerto Lumajang Lawang Batang Ujung Pandang Medan Cirebon Yogyakarta Sidoarjo Langsa, Aceh Tegal Cirebon 15,400 12,800 6,000 4,680 4,560 4,400 4,176 3,940 3,200 3,200 2,880 1,600 1,440 68,276

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Anny Sulaswatty and. Karossi, A.T; Fermentation in Industrial Scale Country Report 1990

Table 1. continued
Commodity and company
F. Beer ** 1. San Miguel Brewery 2. Guinnes 3. Bintang 4. Anker 5. Green Sand G. Glucose and fructose syrups ** 1. PT. Sari Tani Nusantara 2. PT. Raya Sugar Industry 3. PT. Puncak Gunung Mas 4. PT. Trebor Indonesia 5. Delta Chemical Industry 6. PT. Jaya Glucose 7. PT. Indonesian Maltose 8. PT. Super World Wide Food Stuf Indonesia 9. CV. Ebesco 10. PT. Migro Brothers 11. CV. Harum Manis 12. CV. Danaco 13. Firma Industri Kimia Murni
*) Indo Chemical, 1989 & BKPM * *) BBIHP, 1981.

Location of plants

Yearly capacity (ton)

Depend on Market

Bekasi Jakarta Surabaya Jakarta Surabaya

Malang Tasikmalaya Ciracas Jakarta Jakarta Bogor

2,500 2,500 3,000 3,000 350 2,300 2,400 550 450 1,500 900 360 300 20,110

Jakarta Jakarta Jakarta

Table 2. Molasses Production in Indonesia *)

Year 1983 1984 1985 1986 1987

Production (ton) 894,892 847,956 847,700 1,005,260 1,035,184

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Anny Sulaswatty and. Karossi, A.T; Fermentation in Industrial Scale Country Report 1990

Table 3. Projection of Molasses Production in Indonesia *)

Year 1989 1990 1991 1992 1993 1994 Production (ton) 1,284,560 1,348,878 1,416,321 1,487,137 1,561,494 1,639,569

*) Indo Chemical, 1989

The raw material for the production of ethanol, MSG and yeast is sugar cane molasses which is major by-product of cane sugar manufacture. The countrys production of molasses for crop year 1983 - 1987 as shown in Table 2 of which 40% was exported and called Indonesian Blackstrap Molasses, 50% was absorbed by domestic market and 10% was utilized for alcohol manufacture by local distilleries. The projection of molasses production of is presented in Table 3. In 1994 the production will be 1,639,569 ton. Glutamic acid (GA) industry is the first consumer of molasses. Production had begun since 1970 by PT. Ajinomoto Indonesia in Mojokerto, East Java, then developed as illustrated in Table 1. Micrococcus glutamicus or Brevibacterium divaricatum are used as inoculant. Glutamic acid industry in Indonesia seems to have a good prospect because of the by-product such as liquid organic fertilizer and gypsum have enough economic value, beside the raw materials are abundantly produced in Indonesia. Therefore the development of GA and MSG Industries in Indonesia have been fast. Indonesia exports GA and MSG as shown in Table 4 to several countries.

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Anny Sulaswatty and. Karossi, A.T; Fermentation in Industrial Scale Country Report 1990

There are 13 alcohol manufacturers in Indonesia, which actively produce alcohol from molasses. The biggest one is PT. Indo Acidatama Chemical Industry with 15,400 ton per year production, however most of the production is used by it self to produce acid and ester. The development of alcohol industry in Indonesia is not as fast as MSG industry. Nevertheless, the production still increase about 3,5% yearly as shown in Table 5. Being rich in sugar and readily available, molasses is a potential raw material for citric acid production in this country (6). At present, however, cassava solid waste has been used as the raw material instead of cane molasses. The reason for this is probably due to the fact that the technology required for citric acid production from cassava solid waste is cheaper and much simpler than that required when cane molasses is used as the raw material. In adition, cassava solid waste is produced in large quatities and it is relatively cheap and not fully utilized. The production figure of cassava in Indonesia is bout 13,67 million tonnes per year, generating about 1,2 million tonnes of cassava solid waste per year (7). A survey conducted in 1982 indicated that there are six citric acid manufacturers in Indonesia with their total production of 9,130 tonnes citric acid per year. The citric acid industry in Indonesia applied the simplest process, i.e. the solid-state fermentation process (7). In the manufacturing of soy sauce, the fermentation process consists of two consecutive steps, the solid substrate or mold fermentation and the submerged or brine fermentation. In Indonesia, soy sauce known as kecap is widely manufactured and consumed (7). Two kind of soy sauce are well known, the sweetened soy sauce and the salty soy sauce. Most of soy sauce manufacturing is conducted traditionally at home industry scale and although a modern soy sauce factory has been in operation 84

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Anny Sulaswatty and. Karossi, A.T; Fermentation in Industrial Scale Country Report 1990

for more than 10 years. Many studies have been reported on the microbiological and biochemical aspects as well as the recent technology progress in soy sauce manufacturing, but unfortunately relatively little progress has been achieved in applying the relevant biochemical engineering development in the manufacturing of soy sauce in Indonesia. There may be many different reasons for this, scientific and technological, economical, social or managerial in nature (7). Although the industrial scale plants contribute most of the fermentation revenues, Indonesia is rich in the traditional ways of producing food by fermentation. Yeast is used to ferment cassava, which in turn is further processed into cake and fritter called tape. In Bali and east Java, instead of cassava, glutinous rice is used. When the juice it produced is evaporated the solid product is called brem. In Bali, however, it is the liquid that is important to become the famous balinese alcoholic drink. Throughout Java, tempe is well-established protein source; basically it is a soybean cake, a product of soybean fermentation by Rhizopus oligosporus and R. oryzae. Another product using peanut pressed cake as raw material in which Neurospora crassa is the inoculant, is known as red oncom, and when Mucoraceous inoculant is used the black oncom is produced. On the other hand, there are other numerous cottage level fermentation industries throughout the country which provide income to entrepreneur and workers. Cottage level fermentation product include vinegar, nata de coco microbial polysaccharide product in coconut water medium, white cheese fully developed in terms of raw materials, processing methods and product quality. Fermentation process development for both cottage level and large scale industries is expected to contribute significantly to national development. 85

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Anny Sulaswatty and. Karossi, A.T; Fermentation in Industrial Scale Country Report 1990

PROMISING BIOTECHNOLOGY R & D ACTIVITIES Research in old biotechnology has been executed in the universities and research institutes. Lately, tissue culture as a tool for propagating clonally a desired plant species has also been initiated such as for oil palm. Meanwhile, there is a growing interest in Indonesia to enhance research in developing national resources for food, feed, energy and medicines. In the case of antibiotic, since the isolation and description of the first antibiotics, there has arisen a whole generation of semi synthetic penicillins. This substrate is far too complex for total chemical synthesis to be competitive with fermentation. The very fact that antibiotic are subject to microbial degradation has led to the modification of natural antibiotics by microbial enzymes but only a few of such products have become economically important.
Table 4. Indonesian Export of Glutamic Acid and Mono Sodium Glutamate *)
GA Export Tonage 1,140 2,257 2,999 1,010 3,039 1,383 473 US $ 1,757 3,479 4,390 1,343 3,377 1,830 568 MSG Export Tonage 996 1,738 3,027 3,355 7,047 5,610 4,658

Year 1983 1984 1985 1986 1987 1988 1989

*) Indo Chemical, 1989

US $ 1,159 2,323 3,573 3,756 8,032 7,604 5,139

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also incorporate phenylacetic acid into benzyl penicillin (penicillin G) and other precursor into penicillin analogues. The microbial modification of antibiotics, has found success in the enzymatic hydrolysis of penicillin to 6-amino penicillanic acid (6-APA), which is an important substrate for the generation of several semi synthetic and clinically valuable penicillin analogues. Industrially 6-APA, the penicillin nucleus is produced by the hydrolysis of penicillin or benzylpenicillin using strains of microorganisms that produce penicillin acylase in high yield during the antibiotic fermentation or by using immobilized penicillin acylase. Acylase are classified according to the type of penicillin prefentially hydrolysed, some of which are reversible in nature. 6-APA has been used to prepare over fourty thousand semisynthetic penicillins and in some cases the intermediate itself does not have to be isolated, as exemplified by the transformation of benzylpenicillin to ampicillin (5). To have an overview of penicillin fermentation status in Indonesia which still on R & D activities, few authors, mainly from IUC - Biotech ITB, have some report shown in Table 8. They attempted to isolate, characterized and immobilized penicillin acylase from Escherichia coli 1 in polyacrylamide and alginate gel to produce 6-APA. Phenylacetate inducing the activity of penicillin amidase from E. coli B-130 was proven (3).

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Table 5. Alcohol Production in Indonesian *)

Year 1983 1984 1985 1986 1987 1988 Production (ton) 32,600 33,750 30,900 34,200 36,663 44,100

*) Indo Chemical, 1989

Table 6. Indonesian import of Acetic Acid *)

Year 1984 1985 1986 1987 1988 Volume (Ton) 4,279 1,903 8,167 22,692 11,788 Value (US $) 2,226 990 4,250 11,615 6,133

*) Indo Chemical, 1989

Table 7. Indonesian Import of Ethyl Acetate *)

Year 1986 1987 1988

*) Indo Chemical, 1989

Volume (Ton) 3,994 4,435 5,137

Value (Thousand US $) 1,827 2,136 4,323

Direct fermentation, in which antibiotic producing microorganisms are used to synthesize new bioactive compounds in the presence of suitable precurcors or metabolic inhibitors. Penicillium chrysogenum not only synthesize penicillin but will

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Anny Sulaswatty and. Karossi, A.T; Fermentation in Industrial Scale Country Report 1990

Table 8. Activities on the Penicillin Fermentation *)

Product
Penicillin Acylase

Substrate
Bacto Pepton Extract Yeast K2HPO4 KH2PO4 MgSO4 FeCl3 Na-L-Glutamate Phenylacetate Aquadest * Morita Iwata J. Ferment. Tech. 62.217-218,1984 Phenoxy Acetic Pepton Yeast Extract Na-L-Glutamate KH2PO4 K2HPO4 MgSO4 7H2O FeCl3 6H2O Aquadest * Demain 1957 Glucose Ammonia Sulfat * Morikawa & Suzuki Phenyl Acetate Phenyl Acetate Sulfat Glucose

Microorganisms
Escherichia coli 1

Author
Endah T. M. Wirahadikusumah

Penicillin Amidase

Escherichia coli B-130

Adria A. Dessy N. Pringgo S. S. Endah, Roosdiana, Fuafni

Penicillin Acylase

Escherichia coli ATCC9637 ATCC19988 ATCC11105

Penicillin

Penicillin Chrysogenum ATCC9480

Rita R.A. M. Wirahadikusumah

Penicillin G. (Benzyl Penicillin)

Penicillin Chrysogenum

Danu Ariono Herri S.

*) PAU-Biotek ITB, 1990

FUTURE STRATEGIES AND DEVELOPMENT There is an increasing awareness of the importance of biotechnology. Its vast potential as a developmental tool is recognized in a developing agro-based nation such as Indonesia. The Indonesian government supports and encourages biotechnological research and applications wherever appropriate. 89

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In foregoing part of this paper a brief account was given to the potential applications of biotechnology in Indonesia. If an alternative use exists for a particular raw material for which there is an added value, then there is merit in pursuing and supporting research and development that would be ensure its success. This would essentially mean that there is a necessity to increase the existing biotechnology pool in the country to anticipate any development. In conclusion, the impact of biotechnology in agricultural industrial and social development is beginning to make itself felt, and is likely to have a far-reaching consequences in Indonesia. REFERENCES Indo Chemical, PT. Capricorn Indonesia Colsult Inc., Laporan Bisnis Jakarta - Indonesia, No. 38,p.03-30, 16 September 1989. BKPM, Badan Koordinasi Penanaman Modal, List Data Proyek per sektor, sektor Industri Kimia Dasar, periode Januari 1968 - Desember 1988. PAU-Bioteknologi ITB, Kumpulan makalah, Seminar Nasional Bioteknologi Industri, Januari 1990. Proceeding of ASEAN-EEC Seminar on Biotechnology: The Challenges Ahead, The Science Council of Singapore, p. 5 - 9, 1983. I.J. Higgins, D.J. Best & J.Jones, Biotechnology: Principles and Application, Blackwell Scientific Publications, Oxford Lindon Edinburgh, p.132, 1985.

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J.M. Paturau, By-products of the cane sugar industry: an introduction to their industrial utilization, Elsevier scientific Publishing Company, Amsterdam-Oxford-New York, p. 254- 317, 1982. Proceeding On The First Asean Workshop On Biochemical Engineering, Bangkok, Thailand, 25 - 27 September, p. 242, 362., 1985. Balai Besar industri hasil Pertanian, Penelitian pembuatan Gula dengan Sumber Bahan diluar tebu, proyek Survey Aneka Industri, Bogor, 1981. P. Soebijanto, HFS dan Industri Ubi Kayu Lainnya, PT. Gramedia, Jakarta, 1986.

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