Isolation and potency of Actinomycetes from rhizosphere of nutmeg (Myristica fragrans Houtt)

Nutmeg (Myristica fragrans Houtt) is commonly cultivated by people in the forests of Moluccas Islands. This plant grows well on relatively infertile soil types. This is presumably due to the presence of symbiotic microbes in the root of nutmeg. The research aimed to isolate, characterize and test the potential of Actinomycetes from rhizosphere of nutmeg. Soil sample were taken from the nutmeg forest in Ambon Island. The Actinomycetes isolation using humic acid vitamin, continued with yeast malt agar (YMA) media. The testing of antibacterial and antifungal activities using YMA media, while cellulolytic activity, phosphate solubilizing, and xylanolytic activity using carboxyl methyl cellulose, Picovskaya agar, and birchwood agar or oat spelt xylan agar. A total of 12 isolates of Actinomycetes were isolated and dominated by Streptomyces with various types of aerial mycelia. The substrate mycelium looks brown and cream, while the aerial mycelium looks white and gray. These isolates had the highest inhibitory power against Escherichia coli and Fusarium oxysporum with indexes of 16.5 mm and 16.0 mm, respectively. The other isolates have the ability of cellulolytic, phosphate solubilizing, and xylanolytic with indexes 3.26, 3.87, and 1.2, respectively.  The Actinomycetes isolates that were found can be used as starter to improve the biofertilizer formula for nutmeg

Using Lignosellulose Waste as a Xylanase Production Media of Mold Isolated from Rice Straw of Coastal-field

Hemicellulose is one of lignocellulose waste component, so that xylanase is one of importance enzyme of lignocellulose waste biodegradation. Molds as main decomposer lignosellulose waste has enzyme activities higher than yeast and bacteria. The aim of the research is to find mold that have xylanolitic activity using lignocellulose waste as media production. The research consist of isolations and screening mols from coastal-field of watu Ulo Jember, xylanase production using lignocellulose waste and idntification of mold which has the highes xylanase activity. A total of 66 molds isolated from rice straw in coastal-field of Watu Ulo Jember. There were screened for their xylanase activity. In semiquantitatively screen on Oat Spelt Xylan plate, the result showed that 62 have xilanolytic activities. Based on clearing zone production, isolates ESW A1 (3.2), ESW A5 (3.1), ESW C 16 (3.26), ESW D4 (3.0) and ESW D15 (3.21) have xilanase activity index higher than others. Furthermore, quantitative analysis using wheat bran, rice straw and baggase in basic salt Mandel’s modification media showed that xylanase activity of isolate ESW D4 was higher on rice straw 3% as substrate production with activity 2.66 U/mL. Isolate ESW D4 identified as Aspergillus foetidus so that called as Aspergillus foetidus ESW D4. Keywords: rice straw, coastal-field, Aspergillus foetidus ESW-

Characterization of Crude Protease Bacillus sp 31

Bacillus sp 31 was bacteria which produce protease. Characterization of protease from Bacillus sp 31 i.e. pH, temperature, influence of metal ion, enzyme kinetic and enzyme termostability is important to get optimal enzyme activity. Protease activity showed values 146.40 U/ml on pH 9 and optimal temperature 60°C by value. Protease activity increased by addition of 159.50 U/ml Fe2+, but its activity decreased by addition of Mg2+, Cu2+, Ca2+, Al2+, Zn2+ dan Mn2+. Maximal velocity (Vmax) of enzyme-catalysed reactions was 21.32 U/ml with Km 1.5x10-3 mg/ml (Michaels-Menten Kinetic). Protease was very stable at 60°C for 4 hours of incubation and 7 hours of half-time

SCREENING AND IDENTIFICATION ALKALY-CELLULOLYTIC MOLDS FROM RICE STRAW ON COASTAL-FIELD OF WATU ULO JEMBER

About 28 molds were obtained from rice straw on coastal-field of Watu Ulo Jember, were screened for their cellulolytic activities at pH alkaline. In semiquantitatively screening on CMC and Avicel plate at pH 8 showed that 27 isolates have cellulolytic activities. Based on clearing zone in CMC plate, isolates 7, 9, 14, 19 and 24 have higher cellulase (CMC-ase) activities index at pH alkaline. Further, in quantitatively examination using rice straw in Basic Salt Mandel’s modification medium showed isolate 19 (0,60 U/ml) that identified as Aspergillus terreus have higher FP-ase activity than isolate 7  (0,56 U/ml), 9  (0,55 U/ml), 14 (0,53 U/ml) and 24 (0,56 U/ml)

TRANSFORMASI GEN SOSPS1 PADA TANAMAN TEBU OVEREKSPRESI GEN SOSUT1 EVENT 2 MENGGUNAKAN AGROBACTERIUM TUMEFACIENS

Gen SoSPS1 (Saccharum officinarum sucrose phosphate synthase 1) merupakan gen pengkode enzim SPS yang berperandalam biosintesis sukrosa pada organ fotosintesis. Gen SoSUT1 (Saccharum officinarum sucrose transporter1) merupakangen pengkode protein SUT1 yang berperan pada proses transportasi sukrosa dari organ fotosintesis (source) ke organnonfotosintesis (sink). Tujuan penelitian ini adalah untuk mendapatkan tanaman tebu overekspresi ganda yaitu gen SoSPS1dan gen SoSUT1 melalui transformasi gen SoSPS1 pada tanaman tebu overekspresi gen SoSUT1 menggunakan A.tumefaciens. Hasil analisis PCR diperoleh tanaman tebu yang positif overekspresi ganda gen SoSUT1 dan gen SoSPS1sebanyak 4 tanaman pada transformasi ke-1, 3 tanaman pada transformasi ke-2 dan 4 tanaman pada transformasi ke-3.Efektivitas rata- rata transformasi gen SoSPS1 menggunakan A. tumefaciens yang mengandung konstruk plasmid pCL4-SoSPS1 dan dikendalikan oleh promoter RUBQ2 pada tanaman tebu overekspresi gen SoSUT1 sebesar 4,59%

TRANSFORMASI GEN SOSUT1 PADA TANAMAN TEBU MENGGUNAKAN AGROBACTERIUM TUMEFACIENS STRAIN GV 3101 DAN PANGKAL TUNAS TEBU IN VITRO

Gen SoSUT1 merupakan gen pengkode protein sucrose transporter yang memfasilitasi proses transpotasi sukrosa dari jaringan fotosintetik (source) ke jaringan pengguna (sink) pada tanaman tebu. Tujuan penelitian ini adalah mendapatkan tanaman tebu transforman melalui transformasi genetik menggunakan vektor Agrobacterium tumefaciens yang membawa gen SoSUT1. Eksplan pangkal tunas tebu in vitro diinfeksi dengan A. tumefaciens yang membawa konstruk pAct-SoSUT1 dan dilakukan seleksi pada media MS dengan penambahan antibiotik hygromycin.. Tanaman putatif transforman yang telah berhasil melewati proses seleksi dan diaklimatisasi, dilakukan isolasi DNA genom kemudian dianalisis PCR. Hasil analisis PCR dengan pasangan primer 1F/1R hpt II menunjukkan bahwa dari 24 tanaman tebu putatif transforman, didapatkan 15 tanaman tebu positif mengandung gen SoSUT1. Efektifitas rata-rata transformasi gen SoSUT1 menggunakan eksplan pangkal tunas tebu in vitro sebesar 6,8%. Kata Kunci: Agrobacterium tumefaciens, SoSUT1, sukrosa

Growth Pattern and Degradation Activity of Caffeine-degrading Bacteria Consortium

Caffeine-degrading bacteria can be used as agents to degrade caffeine, thereby reducing the concentration of caffeine in organic waste. The decomposition process is carried out by a single bacterium or a consortium of bacteria. Caffeine-degrading bacteria from Sempol, Bondowoso, namely Acinetobacter gerneri KAFS 47, Paracoccus denitrificans KAFS 16 and Pseudomonas plecoglossicida KAFS 34, could be used as a bacterial consortium to promote caffeine degradation. The aim of this study was to analyze associations between caffeine-degrading bacteria isolates, bacterial resistance to antibiotics, growth patterns, and caffeine degradation of a consortium of caffeine-degrading bacteria, and the correlation of bacterial growth with caffeine degradation. The research method used is an analysis of the association between isolates, the development of bacterial consortium growth patterns, and their analysis based on antibiotic resistance, patterning of caffeine degradation, and correlation test (Pearson) of bacterial growth with caffeine degradation. The result of the association test between bacteria showed that the three bacteria had the potential to be used as a consortium of caffeine-degrading bacteria. A. Gerneri, P. denitrificans, and P. plecoglossicida were resistant to the antibiotic cefixime (100 ppm), erythromycin (50 ppm), lincomycin (50 ppm), metronidazole (50 ppm), and sanprima (50 ppm). The growth of the bacterial consortium (54.779 CFU/mL) was higher than that of P. plecoglossicida (49.277 CFU/mL) and lower than that of A. gerneri (93.481 CFU/mL) and P. denitrificans (84.940 CFU/mL) in incubation time of 4 days. However, the consortium of bacteria and P. plecoglossicida were able to degrade caffeine 24 hours faster (3 days) than the other two single isolates (4 days) to degrade 2.5 g/L caffeine in media to 0%. Bacterial growth due to caffeine degradation has a perfect correlation value (>0. 950) and is negative

Identifikasi Aktinomiset Selulolitik dan Xilanolitik Indigenous

Lignoselulosa merupakan penyusun utama dinding sel tumbuhan, sehingga keberadaannya berlimpah di alam. Aktinomiset indigenous yang memiliki aktivitas selulolitik dan xilanolitik ekstraseluler berpeluang sebagai agens biokonversi limbah berlignoselulosa menjadi produk bermanfaat. Penelitian ini bertujuan untuk mendapatkan aktinomiset potensial yang memiliki aktivitas selulolitik dan xilanolitik. Penelitian ini diawali dari isolasi dan pemurnian aktinomiset indigenous asal lahan perkebunan kelapa sawit dan Taman Nasional Bukit Duabelas Jambi.Tahapan selanjutnya adalah penapisan aktivitas selulolitik dan xilanolitik dari aktinomiset, uji pertumbuhan aktinomiset pada mikrokristalin selulosa, dan identifikasi aktinomiset potensial berdasarkan karakter morfologi, fisiologi dan biokimia. Hasil penelitian menunjukkan bahwa aktinomiset isolat S2 yang diisolasi dari lahan perkebunan kelapa sawit mempunyai aktivitas selulolitik dan xilanolitik lebih baik dari keempat isolat lain. Aktinomiset isolat S2 juga mampu tumbuh secara lebih baik pada mikrokristalin selulosa. Aktivitas selulolitik, xilanolitik dan kemampuan tumbuh pada mikrokristalin selulosa dari aktinomiset isolat S2 menunjukkan potensinya sebagai agens pendegradasi material belignoselulosa. Berdasarkan identifikasi morfologi, fisiologi dan biokimia, aktinomiset isolat S2 tergolong dalam genus Streptomyces

Growth of Lactobacillus casei FNCC0900 in Media Based Umbi Porang Plant (Amorphophallus muelleri BI.)

Porang tuber (Amorphophallus muellerii BI.) Is a type of tuber that has a high enough glucomannan content of 67%. Glucomannan is very difficult to digest by humans directly so it takes the role of probiotics. L. casei bacteria FNCC0900 as a probiotic agent capable of utilizing glucomannan as a carbon source for growth. The purpose of this study was to determine the growth pattern and changes in environmental factors, namely the pH value of the probiotic bacteria L. casei FNCC0900 growth medium. The parameters in this study consisted of the highest cell density, generation time and pH value changes in Glucose Yeast Peptone Liquid Media, Porang Boiled Water Media and Porang Flour Liquid Media using the drop plate method which had 4 repeated calculations. Porang Boiled Water Liquid Media has a faster log phase period with a higher cell density than Porang Flour Liquid Media, but the shortest generation time is found in Porang Flour Liquid Media with the highest number of generations. L. casei FNCC0900 bacteria are more able to reduce the pH of Glucose Yeast Peptone Liquid Media compared to porang tuber-based media, so in this case L. casei FNCC0900 can be stated to be able to grow on porang tuber-based media with growth patterns, generation time, cell density and pH value. which varies

KARAKTERSITIK KHAMIR AMILOLITIK DARI BERBAGAI MACAM BUAH

Enam puluh juta ton amilum per tahun di Indonesia memiliki nilai jual yang rendah berdasarkan nilai ekonomi skala internasional. Peningkatan pemanfaatan amilum dapat dilakukan dengan menghidrolisis amilum menjadi glukosa secara enzimatik oleh mikrob amilolitik. Yeast amilolitik memiliki pertumbuhan yang lebih cepat, lebih efektif memecah amilum dari kapang dan dapat digunakan sebagai protein sel tunggal. Penelitian ini bertujuan untuk mendapatkan isolat yeast amilolitik yang berasal dari buah sukun, buah pisang, buah mangga dan buah durian. Hasil isolasi didapatkan 11 isolat yeast. Uji semikuantitatif dengan media SSA terhadap 11 isolat yeast, terdapat 5 isolat yang bersifat amilolitik yaitu  isolat SiJi-P3, SiJi-P5, SiJi-S1, SiJi-S2, dan SiJi-M2. Isolat SiJi-P3 dan SiJi-P5 yang teridentifikasi sebagai Saccarhomyceteae mempunyai indek amilolitik terbesar