Proceeding The 3rd International Conference of Indonesian Society for Lactic Acid Bacteria (3rd IC-ISLAB) : Better Life with Lactic Acid Bacteria: Exploring Novel Functions of Lactic Acid Bacteria.
Table of Contents
List of Supporting Institution...... i Table of Contents...... iii Introduction...... ix Preface from the Chairman of Organizing Committee...... x Preface from the Chairperson of ISLAB...... xi Preface from the Dean of Agricultural Technology Faculty...... xii Summary of Conference...... xiii
Summary Presentation from Invited Speaker The Roles of Probiotics in Gut-brain Axis Communication Leading to Health and Diseases Yuan Kun Lee...... 1 The Advantages and Disadvantages of Lactic Acid Bacteria: Case Study on Tempoyak Fermentation Rindit Pambayun...... 2 Changes in Human Fecal Microbiota Due to Ageing, Nutrition and Chemotherapy and Effects of Probiotic Intervention Alexander.G. Haslberger...... 3 Regulatory Perspectives: Control and Development of Probiotic Foods in Indonesia Roy Sparringa...... 4 Intestinal Microflora and Health in Infants and Children Badriul Hegar Syarif...... 5 The Promising Lactic Acid Bacteria Indigenous Strains for Improving The Quality and Safety of Indigenous Food Product Nyoman Semadi Antara...... 6 Current Methods for the Classification and Identification of Lactic Acid Bacteria Koichi Watanabe...... 7 The Roles of biological resource centers for international cooperation in biotechnology Ken-ichiro Suzuki...... 9 A Functional Probiotic from Kimchi for Atopic Eczema-Dermatitis Syndrome Park Yong Ha...... 11 Diversity of Lactic Acid Bacteria: Biology and Function Fusao Tomita...... 12
Technical Session Papers A1-2 Detection of Bacteriophage Infected-Cell of Lactococcus lactis ssp. lactis C2 Using Acoustic Emission Technique Agustin Krisna Wardani John M. Stencel, and Clair Hicks...... 13 A1-3 Kombucha as Anti Hypercholesterolemic Agent (in Vitro Study using SD rats) Nanik Suhartatik, M. Karyantina, Y. Marsono, Endang S. Rahayu, and Kapti R. Kuswanto...... 22 Full A1-4 Characterization of Bacteriocin From Pediococcus acidilactici F-11 Paper Not Tri Marwati, Eni Harmayani, Nur Richana , and Endang S. Rahayu...... Available A1-5 Biopreservative Study of Starter Culture Pediococcus acidilactici 0094: iii Proceeding The 3rd International Conference of Indonesian Society for Lactic Acid Bacteria (3rd IC-ISLAB) : Better Life with Lactic Acid Bacteria: Exploring Novel Functions of Lactic Acid Bacteria. A1-6 Lactobacillus Fermentum-Like Bacteria of Human Intestine: Structural and Full Functional Properties Paper Not Achmad Dinoto...... Available A2-1 Analysis of bsh Gene From Enterococcus faecium FAIR-E 345 Full Agus Wijaya, Ingrid Specht, Wilhelm H. Holzapfel Paper Not and Charles M.A.P. Franz...... Available A2-2 Survival of L. bulgaricus and S. thermophilus in Coconut Water Based Medium During Frozen Storage Indah Kuswardani, Netty Kusumawati, M. I. Sabrina...... 42 A2-3 Total Lactic Acid Bacteria, Acidity, and Preferences Test of Yoghurt Rice Polish Full Added During 15 Days Refrigerated Storage Paper Not Yoyok Budi Pramono, Dyah Laksmito Rukmi, Heni Rizqiati...... Available A2-4 Microbes Contamination on Organic Vegetables and the Radioresistant of Some Pathogenic Bacteria Harsojo...... 52 A2-5 Survival of Bifidobacteria and Other Selected Intestinal Bacteria in TPY Medium Supplemented With Curcumin as Assessed In Vitro Full , Paper Seyed Davoud Jazayeri, Shuhaimi Mustafa Nassim Naderi, Mohd Yazid Manap, Not Abdul Manaf Ali, and Amin Ismail...... Available A3-1 Fermentation of Peanut Milk by Lactobacillus acidophilus SNP-2 for Production of Non-Dairy Probiotic Drink Giyarto, Titik F Djaafar, Endang S. Rahayu, Tyas Utami...... 58 A3-2 Stability of Lactic Acid Bacteria (Leuconostoc sp) Isolated From Rucah Fish in Liquid Medium) Sri Sumarsih, C. S. Utama, and B. Sulistiyanto...... 67 A3-3 Determination Amylolitic Characteristic of Predominant Lactic Acid Bacteria Isolated During Growol Fermentation, in a Different Starch Medium Composition Widya Dwi Rukmi Putri, Haryadi, Djagal W.M, M Nur Cahyanto...... 72 A3-4 Isolation and Identification of Lactic Acid Bacteria from Local Starter Culture of Full Tape (Ragi Tape) Paper Not Ivannela Kartika, Lindayani, Laksmi H...... Available A3-5 Isolation and Screening Lactic Acid Bacteria From Sayur Asin as Starter for Probiotic Beverages Margaretha Evelyne, Lindayani, Laksmi H...... 82 A3-6 Bacterial Populations during Sorghum Fermentation by Natural Fermentation Full and Using Lactic Acid Bacteria Paper Not Zulianatul Hidayah, Yudi Pranoto, Tyas Utami...... Available B1-1 Dietary Inclusion of Lactobacillus Based Prebiotics, a Natural Solution to Poultry Full Diseases Paper Not Habib Ur Rehman...... Available B1-2 Dietary Supplementation of Lactobacillus-Based Probiotic Help to Alleviate The Full Effects of Heat Stress in Broilers Paper Not Hafsa Zaneb, Saima Ashraf, Umer Sohail, M. S. Yousaf...... Available B1-3 Effect of Lactic Acid Bacteria Probiotics and Antibiotic on Broiler Performance Full Sri Harimurti and Endang S.Rahayu Paper ...... Not Available B1-4 Antibacterial Activity of Kenikir (Tagetes patula L.) Leaf Extracts Against Pathogenic Bacteria and Lactic Acid Bacteria Isolated From Broiler Chickens Septi Nur Hayati, Ema Damayanti, Ahmad Sofyan, dan Hardi Julendra...... 89 iv Proceeding The 3rd International Conference of Indonesian Society for Lactic Acid Bacteria (3rd IC-ISLAB) : Better Life with Lactic Acid Bacteria: Exploring Novel Functions of Lactic Acid Bacteria. B1-6 Isolation and Identification of Lactic Acid Bacteria and Saccharomyces cerevisiae from Natural Sources as Feed-silage Inoculants 97 A. Sofyan, R. Utomo, L.M. Yusiati and Y. Widyastuti...... Full B2-1 Bacterial Lactic Acid Production of Biomass Feedstocks Paper Not Koesnandar...... Available B2-2 Effect of Dietary Pellet Containing of Lactic Acid Bacteria to The Microbial Performance of The Intestine of Broiler Chicks Bambang Sulistiyanto*, Sri Sumarsih and C.I. Sutrisno...... 108 B2-3 The Growth of Lactobacillus salivarius Isolated from Quail Intestine in Soybean Meal Medium Osfar Sjofjan, Umi Kalsum, and Tri Ardyati...... 113 B2-4 The Growth of Lactobacillus Salivarius Isolated from Quail Intestine in Rice Bran Medium Umi Kalsum, Osfar Sjofjan, and Tri Ardyati...... 120 B2-5 Identification of Lactic Acid Bacteria from Quail (Coturnix japonica) Tractus Digestivus and Their Potency to Inhibit Growth of Salmonella typhimurium Tri Ardyati, Osfar Sjofyan, Nur’aini Kartikasari, Pupimadita Tizar A...... 125 B3-1 Development of Indonesian Indigenous Lactobacilli as Probiotic for Diarrheagenic Escherichia coli Full Paper I N. Sujaya, N.P. Desy Aryantini, W. Nursini, K. A. Nocianitri, Y. Ramona, and W Not Redi Aryanta...... Available B3-2 Production of Short Chain Fatty Acid (SCFA) of Sprague Dawley Rats Feeding with Sorbitol Oleic Polyester (SOPE) for Fat Substitute in Diet Agnes Murdiati...... 132 B3-3 Effect of Soygurt Supplemented by Lactobacillus casei subsp rhamnosus on Digesta Profile in Mice Nur Kholis, Kafis Andari Gusti, Jhauharotul Muchlisiyah, Iffah Farisah, Dian Widya Ningtiyas...... 138 B3-4 Evaluation of Indigenous Probiotic Isolate From Rice Bran L. plantarum B2 and Commercial Isolate L. casei in Fermented Rice Bran Media Using In Vivo Full Paper Methode Not Elok Zubaidah, Siti Narsito Wulan, and Anita Kusuma...... Available B3-5 Potency of Lactic Acid Bacteria Isolated From Breast Milk to Prevent Diarrhea Full Caused By Infection of Epec K1.1 Paper Not Lilis Nuraida, Apriliana W. Hartanti, Hana, and Endang Prangdimurti...... Available B3-6 Functions of GI-microbiota and The Quantification of Butyryl-CoA CoA Transferase Genes which Indicate The Butyrate Production Capacity in Full Paper Individuals of Different Diet and Age. Not Berit Hippe, Alexander G. Haslberger...... Available Poster Session Papers P-1 The Production of Soya-Yoghurt Powder by Spray Drying Full Andi Yuni Pratama, Chusnul Hidayat, Indyah S Utami, Ari Wibowo, Lely Novi Paper Not Andriani , Endang S. Rahayu...... Available P-2 Study of The Production of Yogurt Powder by Spray Drying Full M.N.Cahyanto, Eli Mudrik Mafazah, Dedy Widayanto, Irliek Irnastiti, and Endang S. Paper Not Rahayu...... Available P-4 The Use of Carrageenan as A Stabilizer in The Fermentation of Peanut Milk Drink by Lactobacillus acidophilus SNP-2 Palupi Melati Pangastuti1*, Endang S Rahayu2, Tyas Utami...... 147 v Proceeding The 3rd International Conference of Indonesian Society for Lactic Acid Bacteria (3rd IC-ISLAB) : Better Life with Lactic Acid Bacteria: Exploring Novel Functions of Lactic Acid Bacteria. P-5 The Effect of Sugar on Chemical and Sensoris Properties of Keradang (Canavalia virosa) Yogurt 157 Titiek F. Djaafar dan Yeyen Prestyaning Wanita...... P-6 Effects of Tempeh’s Types, Blaching and Skim Addition on Consumer Acceptance of Nuts Tempeh Yoghurt Yeyen Prestyaning Wanita and Siti Rahayu...... 164 P-7 The Characteristics and Sensory Evaluation of Functional Food Based on Cassava Siti Rahayu and Yeyen Prestyaning Wanita...... 171 P-8 Effect of Pediococcus acidilactici F-11 Addition on Antioxidant Activity and Sensory Properties of Probiotic Sweet Potato Ice Cream Retno Utami H and Siti Rahayu...... 179 P-9 Total Count of Lactic Acid Bacteria, Acidity and Total Solid of Banana (Musa paradisiaca sp) Enriched Probiotic Ice Cream B. E. Setiani, S. Mulyani, and Nurwantoro...... 188 P-10 Viability of Lactobacillus plantarum 12A2 during Processing of Probiotic Sweet- corn Ice Cream Netty Kusumawati, Indah Kuswardani, Ignatius Srianta and Eva Setiady...... 194 Full P-11 Tape Ketan Fermentation with the Supplementation of Probiotic Bacteria Paper Not Tenagy, Nurwulan Purnasari, Suparmo, and Endang S. Rahayu...... Available P-12 The Effect of Lactic Acid Bacteria Starter Cultures on the Quality of Yoghurt and Full Gastrointestinal Tract Ecosystem in Mice Paper Not Lovita Adriani and Hendronoto Arnoldus W. Lengkey...... Available P-13 Identification of Lactic Acid Bacteria Strains Expressing Trypsin-Like Activity Full Using 16s rDNA Sequence Paper Not Ika Rahmatul Layly, Is Helianti, and Astutiati Nurhasanah...... Available P-14 The Effect of Growth Medium Removal Prior to Transfer to Production Medium Full towards Medium pH and Trypsin Activity Produced by Lactic Acid Bacteria Paper Not Astutiati Nurhasanah, Dyah Wulansari, Trismilah, and S. Budiasih Wahyuntari...... Available P-15 Isolation and Identification Lactic Acid Bacteria from Raw Beef Meat Full Hendronoto Arnoldus W. Lengkey, Lovita Adriani, Nooremma Sophianie, and Paper Not Roostita L. Balia...... Available Full P-16 Antibacterial Effect of Spices on Fermented Meat Paper Not Purnama Darmadji...... Available P-17 Inhibition Activity of Lactic Acid Bacteria from Pekasam Puka on the Growth of Pathogenic Bacteria Rita Khairina, Edwin Noor Fikri, and Marliana...... 205 P-18 Antimicrobial Activity of Lactic Acid Bacteria Isolated From Bakasang Against Pathogenic Bacteria and Spoilage Bacteria Helen J. Lawalata, Langkah Sembiring, and Endang S. Rahayu...... 212 P-19 Cultivable Lactic Acid Bacteria Isolated from Bekasam (Indonesian fermented fish) and Their Proteolytic and Angiotensin Converting Enzyme Inhibitory Full Paper Activities Not Prima Retno Wikandari, Suparmo, Y. Marsono, Endang, S. Rahayu...... Available P-20 The Quality Improvement of Rusip Product Full Using Starter Culture of Pediococcus acidilactici F-11 Paper Not Arifah Kusmarwati, Tyas Utami, Endang Sri Heruwati, and Endang S. Rahayu...... Available P-21 Effect of Lactobacillus plantarum FNCC 0364 Culture on Microbiological Full Characteristic of Kembung Fish “Peda” Paper Not Murtiari Eva...... Available vi A3-1 Proceeding The 3rd International Conference of Indonesian Society for Lactic Acid Bacteria (3rd IC-ISLAB) : Better Life with Lactic Acid Bacteria: Exploring Novel Functions of Lactic Acid Bacteria. FERMENTATION OF PEANUT MILK BY LACTOBACILLUS ACIDOPHILUS SNP-2 FOR PRODUCTION OF NON-DAIRY PROBIOTIC DRINK Giyarto1, Titik F Djaafar2, Endang S Rahayu3 , Tyas Utami3 1Department of Agricultural Product Technology, Faculty of Agricultural Technology, Jember University, Jember, Indonesia 2Assesment Institute of Agricultural Technology, Yogyakarta, Indonesia. 3Department of Food and Agricultural Product Technology, Faculty of Agricultural Technology, Gadjah Mada University, Yogyakarta, Indonesia. ABSTRACT Production of fermented peanut milk drink by Lactobacillus acidophilus SNP2 using equipments that can be applied in small scale industry was investigated. Peanut milk was extracted using equipments that belong to small scale industry of tofu. Peanut milk inoculated with Lactobacillus acidophilus SNP2 was incubated at 37C for 18 h in incubator that made for small scale industry. After fermentation time, the product contained lactic acid bacteria 9.4 x 108 CFU/ml, with 0.32% titratable acidity and pH 3.86. Sensory evaluation revealed that the panelists prefer the taste of product with addition of 6-10% sugar to that of lower concentration of added sugar. The numbers of lactic acid bacteria showed no marked change in the fermented peanut milk drinks during storage at 4C for 28 days, and still sufficiently high to exert beneficial effects in the host. After storage for 28 days, fermented peanut milk drink contained lactic acid bacteria 6.9 x 10 8 CFU/ml, 0.66 % titratable acid with the pH 3.5. Fermented peanut milk drink can be used as a probiotic carrier. Fermentation of peanut milk drink by Lactobacillus acidophilus SNP-2 for production of non-dairy probiotic drink can be applied to small scale industry. Keywords: Lactic acid bacteria, probiotic, fermented peanut milk INTRODUCTION benefits because of their extreme richness in protein, minerals and essential fatty Peanut (Arachis hypogaea L) is a major acids such as linoleic and oleic acids, source of edible oil and protein meal and which are considered to be highly valuable therefore considered to be highly valuable in human nutrition. It is extensively used in human and animal nutrition. Peanut is in India and other developing countries by also a good source of antioxidant, such as vegetarians and more recently by children p-coumaric acid, that may be contributing allergic to cow milk proteins (Kouane et factors to potential health benefits of their al, 2005). Peanut milk is the water extract consumption (Talcot et al., 2005, Duncan of peanut that is an inexpensive source of et al., 2006). Since peanut has such protein and calories for human potential health benefits, therefore its consumption. Such as soymilk, it is seen as consumption especially in the developing a low-cost substitute for dairy milk for the countries, should be increased. It is developing countries. Being free of necessary to develope of peanut cholesterol and lactose, peanut milk is also processing into other useful and edible a suitable food for lactose-intolerant products. consumers, vegetarians and milk-allergy It has been well known that peanut milk patients. and peanut milk products have nutritional ISBN : 978-979-19546-1-7 Proceeding The 3rd International Conference of Indonesian Society for Lactic Acid Bacteria (3rd IC-ISLAB) : Better Life with Lactic Acid Bacteria: Exploring Novel Functions of Lactic Acid Bacteria. Peanut milk may be produced by soaking fermented with lactic acid bacteria, lactic and grinding full-fat raw peanuts with acid fermented peanut milk may be water to get a slurry, subject to filtration suitable as a probiotic carrier to the host. Many ways of producing peanut milk have been done by various researchers (Beuchat The aims of this study were to study the and Nail., 1978, Bucker et al., 1979, preparation of peanut milk and Isanga and Zhang, 2009). The variation in fermentation of peanut milk drink by L. peanut to water ratio used for peanut milk acidophilus SNP 2 in small scale industry, extractionan affects the peanut milk and to evaluate the cell viability of composition. However, in all cases, this fermented peanut milk drink during lo w cost milk has high protein content. storage at 4C. The current interest in peanut milk and peanut milk products is motivated by the MATERIALS AND METHODS fact that dairy and dairy products are always priced too high for the low income Materials earness. Another factor, no less important, The starter culture of Lactobacillus is the growing awareness of the nutritional acidophilus SNP-2 was a colection of benefits of vegetable proteins in low Food and Nutrition Culture Collection, cholesterol diets by health conscious Center for Food and Nutrition Studies, people (Kouane et al, 2005). Gadjah Mada University, Yogyakarta, Fermentation of peanut milk may serve as Indonesia. The culture was stored in a mixture of 10% glycerol and 10% one possibility that can increase the consumption of this valuable crop, and skimmed milk (1:1) at -20C, and hence improve protein availability and maintained by sub-culturing into sterilized consumption (Sunny-Roberts et al, 2004). MRS media and stored at 4C. Several researchers have studied the The peanut seeds used (Arachis hypogaea fermentation of peanut milk by lactic acid L) were purchased from a farmer in bacteria in laboratory scale (Beuchat and Bantul, Yogyakarta, Indonesia. Care was Nail, 1978, Bucker et al., 1979, Lee and taken to ensure that good quality seeds Beuchat., 1991, Sunny-Roberts et al, 2004, were selected. and Isanga and Zhang 2009). The growth Preparation of Peanut Milk of lactic acid bacteria in peanut milk In the present study, preparation of peanut depends on a number of factors such as milk was performed using the equipment strain of lactic acid bacteria, availability of for soymilk extraction for tofu production nutrition, and fermentation temperature in a small scale industry, called “Tahu and time. Our previous study shows that Kita”. The procedure for peanut milk peanut milk can be fermented using production followed the procedure for Lactobacillus acidophilus SNP-2 to soybean milk preparation for tofu produce peanut milk drink in laboratory production with some modification. scale. L. acidophilus SNP-2 was found to Essentially, whole peanuts were first be resistant to bile salt and low pH, and washed and soaked with the peanut and could be used as probiotic agent water ratio of 1:2 (w/v) for seven hours at (Purwandhani and Rahayu, 2003). room temperature. After draining , the Consumption of tape ketan (fermented peanuts were added with water and ground glutinous rice) supplemented with L. to obtain smooth peanut slurry. The water acidophilus SNP-2 improved the fecal– added for grinding process was about three volunteer microbiota (Rahayu and to four times of the weight of peanut. The Purwandi, 2004). Like soy products slurry then was added with water, to Proceeding The 3rd International Conference of Indonesian Society for Lactic Acid Bacteria (3rd IC-ISLAB) : Better Life with Lactic Acid Bacteria: Exploring Novel Functions of Lactic Acid Bacteria. obtain peanut water ratios of 1:10 and 1: concentration of sugar were kept at cool 20 respectively and put in the extractor room (4C) for sensory evaluation. with agitation speed of 1400 rpm for ten minutes. The peanut slurry then was Determination of Titratable Acidity and moved to the centrifugal separator with a Ph doubled layered cheesecloth on it. The Titratable acidity was estimated using the peanut milk passed through the double method of AOAC (1984), by titration of layered cheesecloth to yield peanut milk. sample with 0.1 N NaOH solution with 1% Peanut milk was then dispended into the phenolpthalein as anindicator. Titratable heating tank at 95C for 5 minutes. The acidity was expressed as percent lactic volume of heated peanut milk and the solid acid, while the pH of the samples was waste were measured, and their proximate measured using a pH meter composition were determined. (ToA/Jenway). Fermentation Peanut Milk Enumeration of Lactic Acid Bacteria Heated peanut milk was added with 10% MRS with CaCO3 was used for the (w/v) sugar instead otherwise stated. It was enumeration of lactic acid bateria. One then cooled and inoculated with 1% (v/v) millilitre of appropriate serial dilutions of of 18 h starter culture of L. acidophilus each samples were pour-plated onto the SNP-2 instead otherwise stated and that media. After 48 h of incubation at incubated at 37C for 18 h in a simple and 37C, the colonies with clear zone that inexpensive incubator that developed for appeared on the plates were counted and small scale industry. Incubator with the the cfu/ml were calculated. size of 90 cm x 60 cm x 45 cm made from plywood. Thermometer, temperature Sensory Evaluation control, three 75 watt lamp and fan for Fermented peanut milk drinks were heat distribution are provided. analyzed for appearance, aroma and taste. Twenty three panelists who had After fermentation, a portion of fermented knowledge of food science and sensory peanut milk drink was distributed in 50 ml quality of fermented drink were used to UV-light sterilized plastic cups and sealed, rate samples on the basis of five points and the rest of the fermented peanut milk hedonic scale. Six samples of fermented was put in the sterile erlenmeyers. The peanut milk drink with various sugar fermented peanut milk drinks either in added concentration were presented to the sealed plastic cup or in sterile erlenmeyer panelists in three digit random number were stored at 4C for 28 days, and sealed cups containing approximately 50 determined the pH, titratable acidity ml of sample. The sensory scores included: (expressed as % lactic acid), and the viable like very much (5); Like moderately (4); counts of lactic acid bacteria. Like slightly (3), Dislike moderately (2); Dislike very much (1). The panelist were For sensory evaluation, fermented peanut also asked to make some milk drink was prepared with various comments/recommendations about the concentration of sugar. Heated peanut milk sensory of appearance and flavor of the was added with various concentration of samples. sugar (0-10% w/v), then inoculated with 1 Statistical Analysis % v/v of starter culture (L. acidophilus The mean values and the standard SNP-2), and incubated at 37C for 18 h. deviations were calculated from the data Fermented peanut milk drinks with various obtained with triplicate trials. Multiple Proceeding The 3rd International Conference of Indonesian Society for Lactic Acid Bacteria (3rd IC-ISLAB) : Better Life with Lactic Acid Bacteria: Exploring Novel Functions of Lactic Acid Bacteria. comparisons of means were carried out by peanut milk was 87.15%. In this study, Duncan multiple range test. peanut milk was prepared for production of fermented peanut milk drink instead of peanut milk yoghurt, therefore we used RESULTS AND DISCUSSION higher peanut and water ratio for preparation of peanut milk. The total solid Extraction of Peanut Milk in the peanut milk does not have to be as Preparation of peanut milk and soymilk in high as for peanut milk yogurt. laboratory scale usually carry out using blender to blend the dehulled seed in Carbohydrate in the peanut milk prepared certain ratio of seed and boiling water. from peanut and water ratio of 1:10 and Filtration is usually performed using a 1:20 were 1.55 and 0.81 respectively double-layered cheesecloth. Preparation of (Table 2). According to Bucker et al., peanut milk for application in a small scale (1979) sucrose was the major fermentable industry, should be carried out using carbohydrate in the peanut milk and simple, inexpensive equipments that are glucose was present at quite low level. readily available. In this study, peanut milk Giyarto (2009) reported that L. acidophilus was produced using equipments that are SNP-2 can grow well in plain peanut milk, available in small scale industry of tofu for but the production of acid is very low. It preparation of soymilk. The minimum means that sugar content in the peanut working volume of extractor is 25 L. For milk supports the growth of that particular peanut to water ratio of 1: 10 and 1:20, the bacteria but the amount of fermentable amounts of peanut and water were 3 kg sugar is not sufficient for the production and 30 L, and 2 kg and 40 L respectively of acid. Thus sugar must be added to (Table 1). After filtration the volume of support the production of acid. heated peanut milks were 27.7 L and 38.2 L, and the weight of solid wastes were The protein level in peanut milks prepared 2.95 kg and 1.80 kg for peanut and water from the peanut and water ratio of 1:10 ratio of 1:10 and 1:20 respectively. Peanut and 1:20 were 2.26% and 1.41% to water ratio for production of peanut respectively. The protein content 1.41% milk varies from one researcher to another. for peanut milk is not far from that of Sunny-Roberts et al.,(2004), used the commercial fermented milk drinks groundnut and water ratio of 5:7 for available in the market. preparation of groundnut milk for The fat content of peanut milks prepared production of a yoghurt–like product. from the peanut and water ratio of 1:10 Peanut milk was prepared by mixing the and 1:20 were 2.69% and 1.48% kernels with water in a ratio of 1:5 for respectively. These values were peanut milk yoghurt (Isanga and Zhang significantly lower than that of prepared by (2009). The ratio of peanut and water on other workers. Previously, other researcher the preparation of peanut milk will affect reported that the fat content of peanut the chemical composition of peanut milk. milks were 4.4% (Bucker et al., 1979), and The proximate composition of peanut milk 7.71%, (Isanga and Zhang, 2009). The is presented in Table 2. Preparation of difference in fat content in peanut milk is peanut milk with peanut and water ratio of mainly due to the difference in peanut and 1:20 produced peanut with higher moisture water ratio used for peanut milk content compared to that with peanut and preparation. The low level of fat in the water ratio of 1:10. Isanga and Zhang peanut milk may be advantage to the (2009) reported that the moisture content keeping quality of fermented peanut milk of peanut milk obtained from peanut and drink as changes of rancidity would be water ratio of 1:5 on the preparation of reduced. Proceeding The 3rd International Conference of Indonesian Society for Lactic Acid Bacteria (3rd IC-ISLAB) : Better Life with Lactic Acid Bacteria: Exploring Novel Functions of Lactic Acid Bacteria. Table 1. Peanut Milk Produced using Equipments fermentation, and the number of for Soymilk Production in Small Scale Industry of 108CFU/ml of lactic acid bacteria was Tofu obtained after 32 h. Hou et al.,(2000) used Component Peanut and Water 1%(v/v) of inoculum for soymilk ratio fermentation that was carried out at 37C 1: 10 1: 20 for 48 h. Lee and Beuchat (1991) used 2% Dry peanut (kg) 3 2 of inoculum for preparation of fermented Volume of water (L) 30 40 Volume of peanut milk (L) 27.7 38.2 peanut milk. Solid waste (kg) 2.95 1.80 The quantity of inoculum normally used is between 3 and 10% (Stanbury et al., Table 2. Chemical Characteristics of Peanut Milk 1995). However, the amount of inoculum used in industry can be lower or higher Peanut and Water ratio than that, due to some reasons. A relatively Component 1:10 1:20 large volume of inoculum culture is used Peanut Solid Peanut Solid to minimize the length of the lag phase. milk waste milk waste Moisture (%) 93.35 76.68 96.21 77.32 Another factor to be considered in the Ash (%) 0.15 0.71 0.09 0.58 determination of the volume of inoculum Protein (%) 2.26 5.57 1.41 5.42 is the economic reason. The higher the Fat (%) 2.69 7.58 1.48 5.86 volume of inoculum, the bigger the vessel Carbohydrat 1.55 9.46 0.81 10.82 size of inoculum preparation. Thus for by different further study, we used 1% inoculum for (%) fermentation of peanut milk drink. Fermentation of Peanut Milk Ratio of peanut and water in the Sugar supplemented peanut milks were preparation of peanut milk affects the inoculated with with 1%(v/v) and concentration of compound in the peanut 10%(v/v) of starter culture of L. milk as seen in Table 2. Although acidophilus SNP-2, and then incubated at extraction of peanut milk using peanut and 37C for 18 h in the simple, inexpensive water ratio of 1:20 produced lower incubator that was developed for small concentration of protein, but protein scale industry. Initially, the amounts of concentration of peanut milk is still higher lactic acid bacteria in the peanut milk than 1%. The use of peanut milk obtained inoculated with 1% (v/v) and 10% (v/v) from the extraction process using peanut 7 starter culture were 3.9 x 10 CFU/ml and and water ratio of 1:10 and 1:20 did not 6 4.8 x 10 CFU/ml respectively (Table 3). give any significant different in terms of However, at the end of fermentation, the pH and viable cells in the end of final numbers of lactic acid bacteria with fermentation (Table 4). Because the 1% (v/v) starter culture was slightly higher purpose of fermentation of peanut milk by than the one with 10% (/v) starter culture. lactic acid bacteria is to produce fermented It means that 1% (v/v) of starter culture peanut milk drink instead of peanut yogurt, was enough to generate high concentration therefore, we used peanut milk that of viable cell at the end of fermentation. In prepared using peanut and water ratio of addition, the present study shows that high 1:20 for extraction process. concentration of starter culture (10%) resulted in the lower pH of peanut milk in This study shows that peanut milk can be the initial fermentation. This condition was prepared using procedure and equipments not favorable for the growth of lactic acid for soymilk preparation in small scale bacteria. Wang et al, (2002) used 0.1% industry of tofu, and simple and (v/v) of an inoculum for soymilk inexpensive incubator can support the Proceeding The 3rd International Conference of Indonesian Society for Lactic Acid Bacteria (3rd IC-ISLAB) : Better Life with Lactic Acid Bacteria: Exploring Novel Functions of Lactic Acid Bacteria. fermentation of peanut milk by lactic acid panelists preferred the aroma, sweetness bacteria. and sourness of fermented peanut milk with addition of 6-10% of sugar. However, Sensory Evaluation panelists dislike the appearance of Table 5. shows the results of the sensory fermented peanut milk drink. When asked evaluation of fermented peanut milk drinks to comment on the appearance, most of with various concentration of added sugar. panelists did not like the appearance Beside for supporting thecell growth and because of poor suspension stability. This acid production during fermentation, findings suggest that if the suspension supplementation of sugar also give some stability could be improved, thus contribution to the sweetness of the fermented peanut milk drink could be fermented peanut milk drink. Fermented more acceptable to potential consumers. peanut milk drink with no addition of Therefore, the suspension stability sugar had the lowest scores in aroma, problem in fermented peanut milk drink sweetness, and sourness. Addition of sugar could be corrected by applying some increased the acceptabilities of panelists to appropriate commercial stablizers. aroma, sweetness, and sourness. The Table 3. pH and Viable Cell Counts of Fermented Peanut Milk with Various Concentration of Starter Culture Starter culture pH Lactic acid bacteria (CFU/ml) concentration (v/v) initial 18 h Initial 18 h 10% 4.89 3.55 3.9 x 107 3.5 x 108 1% 6.45 3.97 4.8 x 106 8.2 x 108 Peanut water ratio = 1:10 (w/v);added sugar 10% (w/v); Incubation temperature 37C Table 4. Fermentation of Peanut Milk by L. acidophilus SNP-2 Peanut water pH LAB (CFU/ml) Appearance ratio * Initial 18 h Initial 18 h 1: 10 6.45 3.97 1.2 x 107 8.2 x 108 Some solid particles 1: 20 6.59 3.86 1.2 x 107 9.4 x 108 Liquid * Peanut and water ratio for peanut milk preparation. Starter culture 1% (w/v));added sugar 10% (w/v); Incubation temperature 37C Table 5. Sensory Characteristics of Fermented Peanut Milk Drink Added sugar Aroma Sweetness Sourness Appearance (%) 0 2.52a 1.39a 1.61a 2.35a 2 2.61b 1.83b 2.17b 2.48a 4 2.83abc 2.26b 2.70c 2.61a 6 2.83abc 3.00c 2.87c 2.61a 8 3.04bc 3.48d 3.04c 2.52a 10 3.22c 3.87d 3.17c 2.65a Survival of Lactic Acid Bacteria during Fermentation of peanut milk at 37C for Storage of Fermented Peanut Milk 1 h, using 1% (v/v) inculum produced Drink fermented peanut milk drink with pH, titratable acidity and viable cell count of Proceeding The 3rd International Conference of Indonesian Society for Lactic Acid Bacteria (3rd IC-ISLAB) : Better Life with Lactic Acid Bacteria: Exploring Novel Functions of Lactic Acid Bacteria. 3.86; 0.32%, and 9.4 x 108 CFU/ml metabolism activity of lactic acid bacteria respectively. During storage the viable during storage. Giyarto (2009) reported the cells counts of L. acidophilus SNP-2 only significant increase in titratable acidity of slightly decreased from 9.4 x 108 CFU/ml sugar supplemented fermented peanut milk at day 0 to 6.9 x 108 CFU/ml and 4.3 x during storage, but not for the plain 108 CFU/ml at day 28 (Table 6). The most fermented peanut milk. Kailasapathy et al., important contributing factors for loss of (2008) also reported the decline in pH of cell viability are decreasing pH during yoghurt during storage. The highest and storage (post-acidification) and the lowest pH at the end of 35-day shelf life accumulation of organic acids as a result study was noted for the plain yoghurt and of growth and fermentation (Shah and the yoghurt with passion fruit. Addition of Jelen, 1990). fruits in the yoghurt will increase the sugar content. This study shows that correlation The initial pH (day 0) of fermented peanut between post-storage pH and titratble milk was 3.86 (Table 6). There was acidity in fermented peanut milk, and the slightly difference between pH at day 0 viability of lactic acid bacteria is affected and day 28 across fermented peanut milk by the presence of added sugar. drink in the sealed plastic cup and in the sterile erlenmeyer. The initial pH (day 0) The present study shows that survival of was 3.86, and the final pH was 3.51. lactic acid bacteria, pH and titratable However, the titratable acidity markly acidity profiles of fermented peanut milk increased during storage. The titratable drink in the sealed plastic cup and acidity in the fermented peanut milk drink erlenmeyer and in the sterile erlenmeyer at day 28 was twice of that at the day 0. during storage are relatively similar. It This increase in titratable acidity was means that sealed plastic cup can be used presumably due to continued fermentation as a packaging for fermented peanut milk by lactic acid bacteria during storage. drink that can be applied in small scale Here, peanut milk was supplemented with industry such as in tofu small scale 10% sugar prior to fermentation. High industry. concentration of sugar supports the Table 6. Characteristics of Fermented peanut milk drink in sealed plastic cup and sterile erlenmeyer during storage at 4C Storage pH Titratable acidity (%) LAB (CFU/ml) time (day) Sealed Sterile Sealed Sterile Sealed Sterile plastic cup erlenmeyer plastic cup erlenmeyer plastic cup erlenmeyer 0 3.86 3.86 0.32 0.32 9.4 x 108 9.4 x 108 3 3.67 3.76 0.50 0.48 8.8 x 108 5.3 x 108 7 3.49 3.56 0.54 0.51 7.7 x 108 4.4 x 108 14 3.85 3.86 0.54 0.54 8.6 x 108 6.8 x 108 21 3.80 3.82 0.60 0.57 7.6 x 108 6.3 x 108 28 3.51 3.51 0.66 0.66 6.9 x 108 4.3 x 108 Starter culture 1% (w/v));added sugar 10% (w/v); Incubation temperature 37C Proceeding The 3rd International Conference of Indonesian Society for Lactic Acid Bacteria (3rd IC-ISLAB) : Better Life with Lactic Acid Bacteria: Exploring Novel Functions of Lactic Acid Bacteria. Overall, fermented peanut milk drinks fermentation of peanut milk. Journal contained greater than 107 CFU/ml of of Food Science. 44, 1534-1538. viable lactic acid bacteria at the end of 28- Beuchat, L.R., and Nail, B. J. , 1978. day shelf life. Thus, at the end of storage Fermentation of peanut milk with time, the fermented peanut milk drink Lactobacillus bulgaricus and L. retained recommeded level of probiotic acidophilus. Journal of Food product. The number of probiotic bacteria Science. 43, 1109-1112. in the food should be at least 107 cfu/ml or Duncan, C.E., Gorbet, D.W., Talcott, S.T., per g at the time of consumption in order 2006. Phytochemical content and to exert beneficial effects in the host antioxidant capacity of water-soluble (Ouwehand dan Salminen, 1998). isolates from peanuts (Arachis hypogaea L.). Food Research International. 39, 898-904. CONCLUSIONS Giyarto. 2009. Production of fermented peanut milk drink by Lactobacillus Preparation of peanut milk can be carried acidophilus SNP-2: evaluation on out using simple equipments in the tofu microbiological, and chemical industry. The simple and low cost characteristics during fermentation incubator has been developed and worked and cold storage.Thesis. Department well for fermentation peabut milk by lactic of Food Science and Technology, acid bacteria for production of fermented Faculty of Agricultural Technology, peanut milk drink. Fermented peanut milk Gadjah Mada University, drink in the sealed cup retained Yogyakarta, Indonesia recommended level (108 CFU/ml) of L. Hou, J.W.,Yu, R.C., Chou, C.C. 2000. acidophilus SNP-2 after four weeks in Changes in some components of cold storage. This procedure for soymilk during fermentation with production of fermented peanut milk drink bifidobacteria. Food Research might be applied to small scale industry. International. 33:393-397. Isanga, J., and Zhang, G., 2009. Production and evaluation of some physicochemical parameters of ACKNOWLEDGEMENT peanut milk yoghurt. LWT-Food This research was financially supported by Science and Technology. 42, 1132- Agricultural Research and Development 1138. Agency, Ministry of Agriculture, Kailasapathy, K., Harmstorf, I., and Indonesia through Research collaboration Phillips, M., 2008. Survival of with University (2008). Lactobacillus acidophilus and Bifidobacterium animalis ssp. lactis in stirred fruit yogurts. LWT-Food Science and Technology. 41, 1317- REFERENCES 1322. Kouane, D., Zhang, g., and Chen, J. 2005. AOAC, 1990. Official Methods of Peanut milk and peanut milk based Analysis, 18th Edition. Association products production. A review. of Official Analytical Chemists, Critical Reviews in Food Science Washington, DC. and Nutrition. 45:405-423. Bucker Jr, E.R., Mitchell Jr, J.H., and Lee, C., and Beuchat, L.R., 1991. Changes Johnson, M.G., 1979. Lactic in chemical composition and sensory qualities of peanut milk fermented Proceeding The 3rd International Conference of Indonesian Society for Lactic Acid Bacteria (3rd IC-ISLAB) : Better Life with Lactic Acid Bacteria: Exploring Novel Functions of Lactic Acid Bacteria. with lactic acid bacteria. a. Peanut has high content of fat. Is it International Journal of Food affect L. acidophilus? Microbiology. 13 (4), 273-183. b. L. acidophilus in peanut milk Ouwehand, A.C., and Salminen, S. J., fermentation uses sugar or peanut as 1998. The Health Effects of Cultured source of Carbon? Milk Products with Viable and Non- c. What is profil sensory of fermented viable Bacteria. International . Dairy peanut milk like? Journal. 8, 749-758. Purwandhani, S.N., and Rahayu, E.S. Answer: 2003. Isolation and selection of a. Ratio water-peanut is 1:20. It means Lactobacillus potential for probiotic that the extract is very dilute, then the agent. Agritech,23: 67-74. fat content of the peanut milk will Rahayu, E.S. and Purwandhani, S.N 2004. very low too. Since it low, it will not Supplementation of Lactobacillus affect L. acidophilus. acidophilus SNP-2 on tape (fermented cassava) and its effect on b. Addition of sugar, increases 2 log human feces. Jurnal Teknologi dan cycle of cell number. It , means that L. Industri Pangan. 15: 129-134. acidophilus can use sugar as C source Shah, N.P., and Jelen, P. 1990. Survival of to growth. Besides uses sugar, L. lactic acid bacteria and their lactases acidophilus also can use carbohydrate under acidic conditions. Journal of of peanut as C source for its growth, Food Science, 55, 506-509. but not for production of lactic acid. Stanbury, P.F., Whitaker, A., Hall, S.J. c. Most panelists can accept the 1995.Principles of Fermentation fermented peanut milk. The problem Technology. Butterworth of fermented peanut milk is the Heinemann, Oxford. appearance. It is caused unstable Sunny-Roberts, E.O., Otulona, E.T., and suspension of fermented peanut milk. Iwakun, B.T., 2004. An evaluation of some quality parameters of a laboratory-prepared fermented groundnut milk. European Food Research Technology. 218:452-455. Talcot, S.T., Passeretti, S., Duncan, C.E., and Gorbet, D.W., 2005. Polyphenolic content and sensory properties of normal and high oleic acid peanuts. Food Chemistry. 90, 379-388. Wang, Y.C., Yu, R.C., and Chou, C.C., 2002. Growth and survival of bifidobacteria and lactic acid bacteria during the fermentation and storage of cultured soymilk drinks. Food Microbiology. 19, 501-508. QUESTION AND ANSWER Question: