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Growth kinetics and Protease Activity 36K Isolates Derived from Mangrove Ecosystem Sediment, Karimunjawa, Jepara (Kinetika Pertumbuhan dan Aktivitas Protease Isolat 36k Berasal dari Sedimen Ekosistem Mangrove, Karimunjawa, Jepara)

ILMU KELAUTAN: Indonesian Journal of Marine Sciences Vol 20, No 3 (2015): Jurnal Ilmu Kelautan
Publisher : Marine Science Department Diponegoro University

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Abstract

Budidaya perikanan laut dan payau menghasilkan sejumlah besar limbah organik yang dibuang ke lingkungan sekitarnya. Sebagian besar limbah organik ini berasal dari sisa pakan, dan feses. Faktor yang menyebabkan terjadinya penurunan produksi budidaya udang adalah munculnya penyakit dan penurunan kualitas air yang dikarenakan adanya akumulasi bahan organik. Penelitian ini bertujuan untuk mengetahui kinetika pertumbuhan dan aktivitas protease bakteri sedimen mangrove dari Karimunjawa - Jepara yang berpotensi sebagai kandidat konsursium probiotik dengan kemampuan aktifitas bioremediasi. Pertumbuhan bakteri isolat 36K memiliki Fase lag (adaptasi) pada waktu kultivasi jam ke-0 hingga jam ke-4. Sedangkan pada jam ke-6 hingga jam ke-30,  isolat 36K  telah  memasuki fase logaritmik (eksponensial) yang dicirikan dengan adanya pertumbuhan yang siknifikan. Pada waktu pertumbuhan jam ke-30 diperoleh nilai OD sebesar 1,939±0,0125 dengan berat kering biomassa bakteri sebesar 1,325±0,043 mg.ml-1, laju pertumbuhan  spesifik (µ) sebesar 0,1446 jam-1, jumlah generasi 8,7609 dan waktu generasi 4,794 jam. Selanjutnya waktu pertumbuhan jam ke-36 hingga jam ke-42, sel isolat 36K  mengalami fase pertumbuhan yang relatif tetap yaitu memasuki fase stasioner. Berikutnya adalah fase kematin pada jam ke-42 hingga jam ke-48, terjadi penurunan laju pertumbuhan yang disebabkan oleh kekurangan materi pertumbuhan seperti vitamin dan unsur mineral. Hasil pengamatan menunjukkan bahwa kadar protein diawal sampai akhir penelitian mengalami penurunan sebesar  97,365%. Nilai laju penurunan kadar protein di media kultur bakteri adalah 0,255±0,005 mg.ml-1jam-1. Produksi enzim protease isolat 36K  dilakukan  ketika  biakan berumur 2 jam hingga 6 jam dengan aktivitas optimum dicapai pada 6 jam masa inkubasi fase eksponensial pertumbuhan bakteri. Kata kunci: sedimen, mangrove, bioremediasi, pertumbuhan, protease  Marine and brackishwater culture produced large number of organic waste, mostly from leftover feed and faeces, which normally released to the nearby water system and becoming source of deseases and reducing water quality.  The study was aimed to determine the growth kinetics and protease activity of 36k isolate from mangrove sediment of Karimunjawa, Jepara which possibly developed into probiotic consortium candidate with bioremediation activity capability. The study was able to describe the growth kinetics and protease activity of 36k isolate which has lag phase (adaptation) on the 0 to the first 4 hours. Meanwhile from 6th to 30th hours isolate 36k entering logarithmic phase (exponential) characterized by significant growth. At the 30th hours period the OD value was 1,939±0,0125, bacterial biomass dry weight of   1,325±0,043 mg.ml-1, specific growth rate (µ) was 0,1446 hours, number of generation 8,7609 dan regeneration time of  4,794 hour. Between 36 to 42 hours the growth of  36k isolate cell was relatively no difference indicating the stationer phase has reached followed by mortality phase between the 42–48 hours where the growth rate was decline due to the limitation of growth material such as vitamins and minerals.  The result also show that protein consentration was significantly decline up to   97,365%. The reduction of of protein content was 0,255±0,005 mg.ml-1.hour-1. The production of enzyme in cultured bacteria was conducted between first 2–6 hours and reach the optimum activity by 6 hours incubation i.e. at the exponential growth phase. The substrate consumption by 36K bacteria isolate was indicated by the decline of reduced sugar with average rate of substrate consumption of 2,0557 g.g-1. Keywords: sediment, mangrove, bioremediasion, growth, protease

Poly-β-Hydroxybutyrate (PHB) Production By Amylolytic Micrococcus sp. PG1 Isolated From Soil Polluted Arrowroot Starch Waste

Indonesian Journal of Biotechnology Vol 19, No 1 (2014)
Publisher : Universitas Gadjah Mada

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Abstract

Poly-β-hydroxybutyrate (PHB) production from amylolytic Micrococcus sp. PG1. Poly-β-hydroxybutyrate(PHB) is an organic polymer, which synthesized by many bacteria and serves as internal energy. PHB ispotential as future bioplastic but its price is very expensive due to glucose usage in PHB industry. Thedevelopment of PHB production using starch as an alternative carbon source has been conducted to reducethe dependence of glucose in PHB production. In this study, amylolytic bacteria from arrowroot processingsite were screened quantitavely based on amylase specifi c activity and PHB producing ability. The result of thestudy showed that among of 24 amylolytic isolates, 12 isolates of them were able to accumulate PHB rangedfrom 0,68-11,65% (g PHB/g cdw). The highest PHB production from substrate arrowroot starch was PG1 andafter optimization resulted in increasing of PHB production up to 16,8% (g PHB/g cdw) 40 hours incubationtime. Based on morphological, biochemical and physiological characters, the PG1 isolate was identifi ed asMicrococcus sp. PG1. Result of the FTIR analysis of produced polymer by Micrococcus sp. PG1 was indicatedas poly-β- hydroxybutyrate (PHB)

Superoxide Dismutase of Micrococcus sp. S2 and Its Involve in Paraquat Detoxification

Indonesian Journal of Biotechnology Vol 12, No 1 (2007)
Publisher : Universitas Gadjah Mada

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Abstract

As an active ingredient of herbicide, paraquat will induce formation of superoxide radicals. The previousresearch succeeded in isolating paraquat degrading bacteria from peat soil, Micrococcus sp. S2, that tolerant to highconcentration of paraquat. An anti-oxidative enzyme, namely superoxide dismutase (SOD, EC.1.15.1.1), wasbelieved to be responsible for the paraquat tolerance. This research was conducted to study the characteristic of theSOD synthesize by Micrococcus sp. S2 and its ability on neutralize superoxide which arise from paraquat reoxidation.To observe the effect of paraquat on Micrococcus sp. S2, the bacteria was grown in 10% Luria Bertani brothmedium amended with several concentrations of paraquat, from 0 (control) up to 100 mg/ml. Within incubationtime of 72 hours, bacterial growth, activity of superoxide dismutase and paraquat residue were analyzed. Theisozymes of superoxide dismutase were distinguished using two kinds of specific inhibitor, namely HO and KCN. 2 2The results showed that paraquat significantly inhibit the growth of Micrococcus sp. S2. The higher paraquatcocentration in the medium caused the higher growth inhibition. However, the bacteria is still survive in the mediumcontaining toxic herbicide, and this ability was suggested related to superoxide dismutase activity in removing thesuperoxide radicals. Analysis using gel electrophoresis indicated that at least three types of SOD isozyme weresynthesized by Micrococcus sp. S2; they were Ferri-SOD (Fe-SOD), Mangani-SOD (Mn-SOD), and the last one wassuspected to be the Cupro Zinc-SOD (CuZn-SOD). The Mangani-SOD was suspected to play an important roles ondetoxifying superoxide which arise from paraquat oxidation.Keywords : Micrococcus sp.S2, paraquat, superoxide dismutase, isozymes

KARAKTERISTIK KONSORSIUM BAKTERI PEROMBAK DIBENZOFURAN DARI SEDIMEN MANGROVE

Jurnal Manusia dan Lingkungan (Journal of People and Environment) Vol 15, No 2 (2008)
Publisher : Pusat Studi Lingkungan Hidup Universitas Gadjah Mada

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Abstract

Dibenzofuran merupakan salah satu senyawa hidrokarbon aromatis polisiklik yang terdiri atas dua cincin benzene yang dihubungkan melalui satu atom oksigen dan satu atom karbon. Dibenzofuran yang mengalami khlorinasi bersifat lebih meracun dari pada senyawa asalnya, dibenzofuran. Penelitian ini bertujuan untuk mendapatkan konsorsium bakteri perombak dibenzofuran dari sediment mangrove serta menyelidiki sifat mikrobiologisnya. Konsorsium dikembangkan dari sediment mangrove menggunakan medium mineral cair yang diperkaya dengan 1000 mg l-1 dibenzofuran sebagai satu-satunya sumber karbon dan energi. Hasil penelitian menunjukkan bahwa konsorsium yang diperoleh mempunyai kemampuan merombak dibenzofuran yang relatif tinggi. Kecepatan perombakan konsorsium terpilih adalah 211,5 mg l-1hari-1. Dari konsorsium terpilih diperoleh lima isolat bakteri. Pertumbuhan konsorsium lebih tinggi dari pada masing-masing isolat atau kombinasinya. Jumlah bakteri dalam konsorsium terpilih mencapai 3,9x1012 CFU ml-1 sedangkan pada masing-masing isolat dan kombinasinya berkisar dari 1,0x108 sampai 7,9x1010 CFU ml-1.

ISOLASI DAN KARAKTERISASI JAMUR PENDEGRADASI ZAT PEWARNA TEKSTIL

Jurnal Manusia dan Lingkungan (Journal of People and Environment) Vol 18, No 2 (2011)
Publisher : Pusat Studi Lingkungan Hidup Universitas Gadjah Mada

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Abstract

Industri tekstil tidak saja menghasilkan sandang yang merupakan kebutuhan primer manusia, tetapi juga mengeluarkan limbah yang berpotensi sebagai penyebab pencemaran lingkungan. Komponen utama limbah industri ini adalah berbagai jenis zat pewarna tekstil. Penelitian ini bertujuan untuk memperoleh isolat-isolat jamur yang mampu mendegradasi beberapa jenis zat pewarna tekstil. Isolasi dilakukan menggunakan metode surface plating di atas medium Potato Dextrose Agar, dan seleksi kemampuan degradasi pewarna berdasarkan atas toleransi terhadap konsentrasi zat pewarna, serta besar dan kecepatan dekolorisasi beberapa jenis zat pewarna. Sebagai parameter awal digunakan enam zat pewarna tekstil. Isolat-isolat unggul kemudian diidentifikasi awal berdasar atas morfologi mikroskopis terhadap miseliumnya. Dalam penelitian ini juga digunakan beberapa kultur murni jamur pembusuk putih sebagai pembanding. Dalam penelitian ini digunakan limbah cair dan padat beberapa industri tekstil dan industri pulp & paper, tanah gambut dari Kalimantan Tengah dan Riau, tanah sekitar Tempat Pembuangan Sampah Akhir, serta tanah seresah hutan. Dari berbagai sumber tersebut diperoleh 101 isolat jamur. Uji dekolorisasi kualitatif terhadap 6 zat pewarna menghasilkan 6 isolat unggul yang mampu mendekolorisasi lebih dari tiga jenis pewarna dengan kecepatan relatif tinggi. Masing-masing isolat unggul memiliki spesifikasi dalam daya dekolorisasi terhadap ke 6 jenis pewarna. Identifikasi awal terhadap isolat unggul menunjukkan bahwa mereka berasal dari genus Aspergillus, Cladosporium, Penicillium dan Stachybotrys. Sedangkan uji terhadap kultur jamur pembusuk putih sebagai pembanding menghasilkan 2 kultur unggul, yaitu: Phanerochaete chrysosporium dan Pleurotus ostreatus. Secara umum kemampuan dekolorisasi isolat-isolat jamur kebanyakan masih di bawah kemampuan kedua kultur murni tersebut, namun beberapa isolat justru memiliki kemampuan lebih tinggi dibandingkan kultur pembanding.

ISOLASI, SKRINING DAN IDENTIFIKASI JAMUR XILANOLITIK LOKAL YANG BERPOTENSI SEBAGAI AGENSIA PEMUTIH PULP YANG RAMAH LINGKUNGAN

Jurnal Manusia dan Lingkungan (Journal of People and Environment) Vol 21, No 3 (2014)
Publisher : Pusat Studi Lingkungan Hidup Universitas Gadjah Mada

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Abstract

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.

INFLUENCE OF PARAQUAT HERBICIDE ON SOIL BACTERIA Rhizobium sp.

Jurnal Manusia dan Lingkungan (Journal of People and Environment) Vol 8, No 2 (2001)
Publisher : Pusat Studi Lingkungan Hidup Universitas Gadjah Mada

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Abstract

Pencemaran pestisida merupakan salah satu masalah lingkungan yang menyebabkan gangguan terhadap organisme tanah. Paraquat adalah bahan aktif beberapa jenis herbisida yang banyak diaplikasikan di lahan gambut dan lahan pertanian tadah hujan. Studi ini dilakukan untuk mengetahui pengaruh herbisida paraquat terhadap bakteri Rhizobium sp. Tiga puluh lima strain Rhizobium sp. telah diuji dengan menggunakan teknik difusi cakram kertas (paper disc). Sebagian strain adalah hasil isolasi dari ranah, bintil akar tanaman Ieguminosa dan inokulum leguminosa (Legin - Iegume Inoculum). Enam strain lain aclalah bakteri Rhizobium japonicum. Hasil penelitian menunjukkan hahwa paraquat memiliki daya hambat terhadap hakteri Rhizohium sp. Sebanyak 17,14% (enam strain) dari seluruh strain yang diuji, tidak mengalami penghambatan sampai konsentrasi paraquat 400 ppm. Bakteri ini memiliki prospek bagus untuk digunakan sebagai inokulum rhizobium terutama di lahan pertanian yang telah tercemari herbisida, khususnya yang mempunyai bahan aktif paraquat. Sebagian besar strain yang digunakan (82,86%) terhambat oleh 20 ppm parakuat dan daya hambat tersebut makin besar seiring dengan meningkatnya konsentrasi paraquat. Mengingat makin meluasnya pemakaian herbisida berbahan aktif paraquat di Indonesia dan peran Rhizobium dalam fiksasi nitrogen, hasil penelitian ini fiksasi memiliki arti penting, terutama bagi petani agar berhati-hati dalam pemakaian pestisida.

Herbisida Parakuat dalam Lahan Gambut: II. Pengaruhnya terhadap Pertumbuhan dan Hasil Kedelai

Jurnal Perlindungan Tanaman Indonesia Vol 7, No 1 (2001)
Publisher : Universitas Gadjah Mada

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Abstract

Utilization of peat soil for agriculture faces several constraints, such as low soil fertility and pH value. No-tillage agricultural system in peat land needs application of herbicides, for example herbicides with paraquat as active agent. This research was conducted to study the influence of paraquat on the growth and yield of soybean in peat. Peat soil from Central Kalimantan Province was obtained for this study. Due to the requirement of the plant, the soil was treated with lime and NPK-fertilizers. The results showed that paraquat significantly inhibited vegetative growth and decreased soybean production. These phenomena were found in early and advanced decomposed peat soil. Enhancement of vegetative growth and yield were detected when peat was treated with lime and or fertilizers. Liming treatment also supported the formation of root nodules. Inspite of increasing the growth of soybean, lime and fertilizers addition in peat was able to neutralize the negative effect of paraquat on the growth and yield of soybean. The best growth and yield of soybean were found in early decomposed peat soil, i.e. fibric peat soil.Key words: paraquat, peat, soybean

Paraquat Toxicity on Root Nodule Formation on Macroptiliuma tropurpureum Urb. and Its Corelation with Population of Rhizobium sp.

Jurnal Perlindungan Tanaman Indonesia Vol 10, No 2 (2004)
Publisher : Universitas Gadjah Mada

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Abstract

This study was designed to investigate the paraquat toxicity toward root nodulation by Rhizobium on Macroptilium atropurpureum as an indicator plant. The legume was grown in Thornton medium treated with several concentrations of paraquat and inoculated with R.japonicum 143 (Rj-143) or Rhizobium sp. C-1.1. These bacteria represent cross-inoculation of soybean and cover-crops legumes, respectively. Nodule formation and Rhizobium population were measured periodically. At the end of planting time, nitrogenase activity of the nodules was analysis based on ARA (Acethylene Reduction Analysis) method. The results showed that nodules in plants inoculated with Rhizobium without addition paraquat, were formed within four weeks. There was no nodulation when paraquat was added. Paraquat was toxic to the plant, causing chlorosis, stunting, drying of the plant tissues, and death. The symptoms were detected at the second week after planting time. Paraquat also decreased Rhizobium population from 10^6 to 10^2 or 10^1 CFU/mL at 40 and 100 pp, respectively. These results depicted that paraquat disturbed the plant before nodulation, and at the same time Rhizobium populatin decreased until below minimal population required for nodulation. Therefore, the process of nodulation was disturbed, and in some treatments there was nodulation. It was concluded that paraquat was toxic to both plant and the Rhizobium, which cause nodulation failure.

Paraquat Toxicity on The Growth of Rhizobium sp. in A Synthetic Medium

Jurnal Perlindungan Tanaman Indonesia Vol 8, No 2 (2002)
Publisher : Universitas Gadjah Mada

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Abstract

Toxicity of paraquat on the growth of several strains of Rhizobium sp. in Yeast Extract Mannitol medium was studied. Various concentrations of paraquat 100, ranged from O (control) to 100 ppm were applied. Qualitative examination was done using paper disc diffusion technique, and the quantitative examination was conducted based on the change in cell density in medium measured by plate count method.Qualitative data showed that effect or paraquat was species specific. Some strains of Rhizobium sp., namely Rhizobium sp. strain T-37 and QF, were tolerant to paraquat until100 ppm, but other strains were sensitive to paraquat, especially at high concentration.Quantitative examination to the sensitive strains shows that higher concentration of paraquat caused higher toxicity to the growth of Rhizobium. Rhizobium sp. strain G-69 and  G-182.paraquat addition at 100 ppm slightly decreased cell density from 10 to 10 CFU/mL. Rhizobium japonicum strains 143 and KS were tolerant to 20 and 40 ppm of paraquat; their cell density increased from 10^6 to a level of 10&7 or 10^8 CFU/mL depend on the strain. This level was not significantly lower than those in medium without paraquat, in which maximal population density reached to 10^8 or 10^9 CFU/mL. Addition of higher paraquat concentration damaged the cell of these strains, and caused population density increased specifically to a level of 10^1 CFU/mL. The data which showed that paraquat was toxic to Rhizobium sp. were important growth inhibitor, because growth inhibition of these bacteria may influence the formation of root nodule on leguminous plants, and in turn will decrease the yield. Due to widely applied paraquat in agricultural and plantation systems, and the role of Rhizobium in nitrigen fixation, these results are important for minimizing the impact of paraquat application.