Isolasi dan Karakterisasi B-1,3-glukanase Akar Semai Tusam (pinus merkusii jungh. Et de vriese) yang Berasosiasi dengan Fungi Ektomikorisa: Isolation and characterization -1,3-glucanase from tusam seedling roots (pinus

ABSTRACT Association between tusam (Pinus merkusii Jungh. et de Vriese) seedling roots with ectomycorrhiza fungi was expected to increase the activity f3-1,3-glucanase level in the plants. This enzyme has potency to protect seedling from soil borne fungal pathogens by degrading the fungal cell walls. The objectives of this research were to isolate and characterize 0-1,3-glucanase from tusam seedling roots associated with ectomycorrhiza fungi. Crude protein was isolated with ammonium sulfat precipitation and then purified by gel filtration chromatography and characterize molecular weight, temperature and pH for optimum the activity.- The enzym frdm tusam seedling roots associated with ectomycorrhizal fungi, designated as GLUC15 have 15 kD molecular weight, 30° â 40° C temperature and pH 5 â7 for optimum activity. Key words: f3-1,3-glucanase, tusam, fungi, ectomycorrhiza INTISARI Asosiasi antara akar tusam dengan fungi pembentuk ektomikorisa diduga mengimbas adanya enzim13-1,3-glukanase. Enzim ini memberikan kontribusi terhadap ketahanan semai tusam dalam menghadapi fungi patogen melalui aktivitas enzim tersebut dalam mendegradasi dinding sel fungi yang mengandung glukan. Penelitian ini bertujuan untuk melakukan isolasi dan karakterisasi 0-1,3-glukanase akar semai tusam yang berasosiasi dengan fungi pembentuk ektomikorisa. Crude protein diisolasi melalui tahapan pengendapan ammonium sulfat 70% dan kemudian dipisahkan menggunakan kromatografi gel filtrasi. 13 - 1 ,3-glukanase yang diperoleh dikarakterisasi berat molekul, pH, dan suhu optimal aktivitas enzim. Enzim 13-1,3-glukanase dari akar semai tusam yang berasosiasi dengan fungi pembentuk ektomikorisa, yang kemudian disingkat GLUC15 merniliki berat molekul 15 kD, kurang tahan pemanasan dengan suhu optimal aktivitas 30° -40° C, bekerja pada kondisi asam dengan aktivitas pH optimal 5 -7

ISOLASI DAN SELEKSI BAKTERI AMILOLITIK PENYEBAB KEMASAMAN PADA TEPUNG SAGU BASAH HASIL PENYEDIAAN SECARA TRADISIONAL

Kemasaman pada tepung sagu basah disebabkan karena adanya asam organik hasil perombakan tepung yang dilakukan oleh bakteri. Penelitian ini bertujuan untuk mengisolasi bakteri amilolitik penyebab kemasaman berdasarkan seleksi kemampuan amilolitik dan penghasilan asam organiknya. Sumber isolat diambil dari tempat penyediaan tepung sagu basah di Jayapura dan sekitarnya. Kemampuan amilolitik ditentukan dengan uji amilolitik menggunakan reagen iodium sedangkan analisa asam organik dilakukan menggunakan perangkat gas kromatografi. Isolasi menghasilkan 114 isolat bakteri amilolitik penghasil asam organik. Sebagian besar isolat amilolitik mampu menghasilkan asam laktat dengan kadar lebih tinggi dibanding empat jenis asam organik lainnya. Meskipun demikian bau masam pada tepung sagu bukan hanya disebabkan oleh kehadiran asam laktat saja tetapi juga oleh adanya asam asetat, propionat dan butirat sebagai produk fermentasi bakteri amilolitik

Isolasi, Skrining Dan Identifikasi Jamur Xilanolitik Lokal Yang Berpotensi Sebagai Agensia Pemutih Pulp Yang Ramah Lingkungan (Isolation, Screening and Identification Xylanolytic Local Fungi That Potentially as Pulp Bleaching Agents)

Xilanase merupakan enzim yang berfungsi luas dalam bidang industri. Xilanase digunakan sebagai perlakuan awal proses pemutihan kertas di industri pulp dan kertas sehingga dapat mengurangi penggunaan senyawa klorin yang berbahaya bagi lingkungan. Xilanase yang cocok digunakan dalam industri pulp dan kertas seharusnya bebas dari aktivitas selulase. Jamur merupakan salah satu kelompok mikrobia yang mampu menghasilkan xilanase. Penelitian ini bertujuan untuk memperoleh isolat jamur unggul lokal penghasil xilanase dari tanah yang diasumsikan memiliki kandungan xilan tinggi. Tanah di sekitar industri pulp dan kertas; hutan akasia di Kab. Muara Enim dan Ogan Ilir, Sumatera Selatan; hutan Wanagama, Yogyakarta; penggergajian kayu di kota Palembang dan Yogyakarta serta TPA Palembang digunakan sebagai sumber isolat jamur. Berdasarkan skrining awal dalam media basal xilan agar diketahui bahwa dari 111 isolat jamur yang diperoleh, sebagian besar mempunyai potensi menghasilkan xilanase, akan tetapi hanya 12 isolat yang mempunyai kemampuan xilanolitik tinggi. Skrining selanjutnya dilakukan pada media basal xilan cair menunjukkan bahwa jamur yang diidentifikasi sebagai Chaetomium globosum, Penicillium simplicissimum, Aspergillus tamarii dan Monocillium sp. berpotensi unggul dalam menghasilkan xilanase dibandingkan isolat lainnya berdasarkan aktivitas enzim spesifiknya. Keempat jamur tersebut diketahui juga memiliki aktivitas lignolitik dan selulolitik. Oleh karena itu, xilanase yang diproduksi ke empat jamur tersebut berpotensi dikembangkan sebagai agen pemutih pulp

Kemampuan Isolataktinomisetes Menghasilkan Enzim yang Dapatmerusak Kulit Telur Nematoda Puru-Akar Meloidogyne Spp.

Soil microbes including actinomycetes are known to produce various hydrolytic enzymes and antibiotics that can be used as biological controlling agents nematode. Therefore, surveys conducted in several areas in Yogyakarta, Central Java and East Java, to search for actinomycetes with chitinolytic, proteolytic, and chitino-proteolytic activity. Isolation of Actinomycetes produced 84 isolates, and most was obtained from shrimp head waste (26 isolates). After the selection based on their ability to hydrolyze chitines and protein in the medium, those whith the highest chitin and protein hydrolysis activity, are consecutive PSJ 27, TL 8, and TL 10 isolates. Test results of crude enzyme produced by selected isolates against root-knot nematode eggshell, showed that the isolates that have chitino-proteolytic activity (TL 10), is a highly effective isolate in damage eggshell. There are three types of damage to the nematode eggs. In the young eggs, crude enzyme preparation causing damage on vitelline and chitin layers. In the older eggs, preparation of crude enzyme cause premature hatching. Sebagian mikrobia tanah, termasuk aktinomisetes, diketahui mampu menghasilkan berbagai enzim hidrolitik dan antibiotik yang dapat dimanfaatkan sebagai agens pengendalian hayati nematoda. Oleh karena itu,survei dilakukan di beberapa daerah di Yogyakarta, Jawa Tengah, dan Jawa Timur untuk mencari aktinomisetes yang mempunyai aktivitas kitinolitik, proteolitik dan kitino-proteolitik. Isolasi aktinomisetes menghasilkan 84 isolat, dan yang terbanyak diperoleh dari limbah kepala udang (26 isolat). Setelah dilakukan seleksi berdasarkan kemampuannya menghidrolisis kitin dan protein dalam medium, yang mempunyai aktivitas hidrolisis protein, kitin, protein dan kitin tertinggi berturut-turut adalah isolat PSJ 27, TL 8, dan TL 10. Hasil uji enzim kasar yang dihasilkan isolat terpilih terhadap Perusakan kulit telur nematoda puru-akar menunjukkan bahwa isolat yang memiliki aktivitas kitino-proteolitik (TL10) merupakan isolat yang sangat efektif dalam merusak kulit telur.Terdapat tiga tipe kerusakan pada telur nematoda. Sediaan enzim kasar menyebabkan kerusakan atau terkoyaknya lapisan vitelin dan lapisan kitin pada telur muda. Pada telur yang sudah tua, sedíaan enzim kasar menyebabkan pecahnya lapisan kulit telur yang menyebabkan penetasan yang prematu

Isolation and Lignocellulolytic Activities of Fiber-digesting Bacteria From Digestive Tract of Termite (Cryptothermes SP.)

The objectives of this study were to obtain the fiber-digesting bacteria isolates from termitedigestive tract and to determine the optimum conditions of growth and production of cellulase, xylanaseand ligninase enzyme of isolate. The first study was conducted to isolate and select the fiber-digestingbacteria from the digestive tract of termites based on the highest activity of cellulolytic (S), xylanolytic(X) and lignolytic (L). The second study was optimation of the growth conditions of bacteria and theenzyme production due to effect of rice straw substrate and nitrogen. The material used were dry woodtermites, rice straw, and culture medium. The design used was a completely randomized factorial design,in which the first factor was rice straw substrate (1, 2, and 3% W/V), while the second factor wasnitrogen (0.1, 0.2 and 0.3% W/V). Variables measured were cellulase, xylanase and ligninase activities.Results of the first sudy showed that the isolates obtained consisted of 3 types, those were cellulolyticbacteria (S1, S2, and S3), 3 types of bacteria xylanolytic (X1, X2, and X3) and 3 types of bacteria lignolytic(L1, L2, and L3). Meanwhile, results of the second study showed that isolates of S2, X3, and L1 had thehighest activity, those were 1.894 U/mL, 1.722 U/mL and 0.314 U/mL, respectively. In conclusion, the addition of 1% level of rice straw substrate and 0.3% of nitrogen showed the highest enzyme activity oncellulase, xylanase and ligninase

Skrining bakteri asam laktat penghasil bakteriosin dari daging dan produk olahannya=(screening of bacteriocin producer lactic acid bacteria from meat and its products)

ABSTRACT Lactic acid bacteria (LAB) which naturally occur in meat and meat products have been isolated and screened for their ability to produce bacteriocin. The objective of this research was to obtain the potential bacteriocin producer of lactic acid bacteria which could be used as food bio-preservative. Source of lactic acid bacteria used in this study were beef chicken flesh, \u27vacuum packaged\u27 sausage and sliced meat obtained from traditional market or department store. Ten grams of each samples was put onto five different enrichment media, i.e., TGE (tryptoneglucose-yeast extract) pH 5 plus 3% NaClMRS (deMan Rogose Sharpe) pH 5,5TGE broth pH 5,5TGE buffer broth pH 5,5and TGE broth plus Tween 80 & 1% Naazida pH 6,0, incubated for 24-71 hours to stimulate the growth of lactic acid bacteria. Different enrichment media were used to stimulate the growth of strains belong to each genus, since the nutritional and environmental requirement for optimum growth were suggested to be genera-dependent. Screening of LAB bacteriocin producer was carried out by dilution -pour plate methods (culture from each enrichment medium) followed by overlay using the indicator strains. Indicator strains used in this study were Lactobacillus plantarum NCDO 955, Pediococcus acidilactici LB-42, Leuconostoc mesenteroides LY, and Enterococcus faecalis MI. Colonies showing growth inhibition to indicator (indicated by clear zone) were isolated and purified. Isolates were then characterized based on Gram, catalase, shape and arrangement of cell, type of fermentation, effect of temperature to the growth and acid production from several carbon sources. From the primary screening (dilution - pour plate âoverlay), 30 strains belong to Lactobacillus, Leuconostoc, Streptococcus and Enterococcus which suspected to produce antimicrobial substance were obtained. However, based on the confirmation lest (dUsion method), only three (3) strains were identified to produce bacteriocin. i. e. Leuconostoc mesenteroides SM 22, SM 32, and SM 46. In this study, Leuconostoc mesenteroides SM 22 was selected for food application. Bacteriocin of Leuconostoc mesenteroides SM-22 was able to inhibit the growth of psychrophilic bacteria naturally occur in meat and shrimp kept at refrigerator. Microbial population of raw meal with the initial number of about 3x104 CFU/g decreased one log cycle after treated with bacteriocin, and this number maintained less than 105 CFU/g after storage raw meat at refrigerator for five days. On the other side, microbial population of raw meat with no bacteriocin treatment increased to 106 CFU/g after 4 days kept at refrigerator. In the case of shrimp, washing raw shrimp with cold water could reduce the population of bacteria about one log cycle, followed treatment with bacteriocin, this populationincreased very slowly and still less than 105 CFU/g after 5 days storage at refrigerator. While without any treatment, microbial population of raw shrimp which initially about 3x105 CFU/g rapidly increased to 106 CFU/g after 3 days. This data showed that Leuconostoc mesenteroides SM-22 was a potential bacteriosin producer and can be applied as bio-preservative for cold storage fbod. Keywords : Bakteri Asam Laktat, Daging, Bakteriosi

Isolation and Selection of Rhizobium Tolerant to Pesticides and Aluminum From Acid Soils in Indonesia

Application of Rhizobium as inoculum in acid soil requires specific characters, namely high tolerance to pesticide residues, soil acidity, and high concentration of Aluminum. This study was conducted to isolate Rhizobium having these characters. Inspite of acid soils from Kalimantan, Sumatra, Sulawesi and Java; root nodules of legumes planted in those regions were used as source of isolates. Rhizobial isolation was done using direct isolation andenrichment technique. A paper disc diffusion technique was used in selecting tolerance to pesticides. The selected isolates were examined the tolerance to pH, Al, and ability to form root nodule with soybean. From soil analysis, it could be seen the correlation between pH value and Al concentration. It means that the lower pH value the higher Al concentration. The number of Rhizobium isolates and its tolerance to paraquat was depended on soil type. From 173 strains of isolated Rhizobium, 24 strains were tolerance to pesticides and Aluminum. They were able to grow in wide range of pH, namely 3 – 8, or some of them in 5 - 8. Around 92% of the selected bacteria could form root nodules with soybean plant in different number and size. Hopefully, these isolates can be applied in the pesticide polluted agricultural lands, especially in acid soils with high concentration of Al, and it can also increase soybean production

Isolasi dan Karakterisasi Kitinase Akar Tusam (Pinus merkusii Jungh. et de Vriese) yang Bersimbiosis dengan Fungi Ektomikorisa: Isolation and characterization chitinase in tusam (Pinus merkusii Jungh. et de Vriese) roots

ABSTRACT The experiment was aimed to detect the activity and characterize the chitinase of tusam root during ectomycorrhizal symbiosis. Tusam inoculated with tusam stands soil from Kaliurang as fungi inocula. Crude proteins were isolated from 4, 6, and 8 weeks age of Chitinase activity and isoform were detected using glycol chitin as a subtrat. The .enzyme was purified by ammonium sulfa( precipitation, dialysis, followed by gel filtration chromatography. The results showed that tusam produced chitinase with a molecular weight of approximately 52 kDa. The optimum activity was at pH 5 and temperature of 30°C. Key words: isolation, characterization, chitinase, ectomycorrhizal, tusam INTISARI Penelitian ini bertujuan untuk mengetahui akti vitas dan karakterisasi kitinase yang terjadi waktu akar tusam bersimbiosis dengan fungi ektomikorisa. Semai tusam diinokulasi dengan fungi ektomikorisa, yang berasal dari tanah dibawah tegakan di Kaliurang. Protein kasar diisolasi dari semai tusam pada umur 4, 6, dan 8 minggu. Aktivitas kitinase dan isoform dideteksi menggunakan substrat glikol kitin. Pemisahan protein diawali pengendapan dengan ammonium sulfat, dialisis, dilanjutkan dengan kromatografi gel filtrasi. Hasil penelitian menunjukkan bahwa tusam memproduksi kitinase yang mempunyai berat molekul 52 kDa, suhu 30°C, dan pH optimum 5. Kata kunci: isolasi, karakterisasi, kitinase, ektomikorisa, tusa

Polyphasic identification of amylolytic bacteria producing bioplastic Poly-β-hydroxybutyrate (PHB)

The final goal of this study is to make a modern systematic-based inventory of amylolytic bacterial isolates producing of bioplastic Poly-β-hydroxybutyrate (PHB) from sago starch substrate. The identity of three local bacterial isolates was examined in this study, using a polyphasic approach. A data set based on phenotypic characteristics, namely morphological, physiological, biochemical and chemical character, namely whole cells protein profiles using SDS-PAGE method, together with phylogenetic studies based on 16S rRNA sequences was used to identified by polyphasic approach. Phenotypic characteristics of 3 local bacterial isolates and 4 reference strains to members of genus Bacillus was analyzed by numerical analysis using MVSP 3,1 program to determine the value of similarity. Based on the preliminary characterization of the profile matching method showed that the three isolates of bacteria producing PHB namely PSA10, PPK5 and PPK6 are members of the genus Bacillus. The results of numerical analysis based on phenotypic characteristic and chemical character of the three bacterial isolates producing PHB with reference strains showed that the PSA10 isolate bacterial identical with Bacillus megaterium, PPK5 isolate identical with Bacillus subtilis and PPK6 isolate identical with Bacillus cereus, and these results also support by the molecular phylogenetic analysis. Therefore, the polyphasic taxonomy is an effective approach to uncover the identity of the novel bacterial isolates