PRODUCTION AND CHARACTERIZATION OF CRUDE INTRACELLULER PHYTASE FROM RECOMBINANT BACTERIA pEAS1AMP

This research was aimed at producing a crude intracellular phytase characterized from recombinant bacteria. The recombinant bacteria (pEAS1AMP) was produced by way of transforming pET-22b(+) +pEAS1 into competent E. coli BL21 and E. coli BL21(DE3) cells. Crude intracellular phytase production was induced using 1,5 mM Isopropyl-β-D-thiogalactopyranosid (IPTG). Recombinant bacteria product and enzyme activity test followed the Sajidan method. E. coli BL21(+)pEAS1 and E. coli BL21 (DE3)(+)pEAS1 recombinant bacteria showed growth after 20 hours and 10 hours of transformation. Phytase activity of E. coli BL21 (DE3)(+)+pEAS1 showed higher than those of E. coli BL21(+)+pEAS1. Crude intracellular phytase of pEAS1AMP recombinant bacteria has an optimum activity at pH 5, 40o C, incubation period of 60 minutes, substrate concentration of 2%, molecular weight (MW) of 47.3 kDa, Km = 15.91 υM and Vm = 2.41 υM/second. Mg2+ acts as a cofactor but Fe3+ (10-4 M) acts as an inhibitor. Keywords: bacteria recombinant pEAS1AMP, competent cells, crude intracellular phytas

Improving Environmental Attitude and Etichs Trough Subject Specific Pedagogy

Improving Environmental Attitude and Etichs Trough Subject Specific Pedagog

Improving Environmental Attitude and Etichs Trough Subject Specific Pedagogy

Improving Environmental Attitude and Etichs Trough Subject Specific Pedagog

Kimpul (Xanthosoma spp.) characterization based on morphological characteristic and isozymic analysis

Nurmiyati, Sugiyarto, Sajidan. 2009. Kimpul (Xanthosoma spp.) characterization based on morphological characteristic and isozymic analysis. Nusantara Bioscience 1: 138-145. This research is aimed: (i) to know the variety of kimpul (Xanthosoma spp.) based on morphological characteristics and isozymes analysis; (ii) to know the correlation between its genetic space based on morphological characteristics and its genetic resemblance based on isozymes-banding pattern. This research results were analyzed and described by descriptive qualitative methods. Morphological observation was carried out in sub-District of Galur, Lendah and Girimulyo, Kulonprogo District, Yogyakarta. Morphological data of the kimpul plant was explored descriptively and then made dendogram. Data of isozymic banding pattern were analyzed quantitatively based on the appearance of the band on the gel, and qualitatively based on the thickness of the band formed, and then made dendogram. The correlation, between its genetic distance based on morphological characteristics and its genetic resemblance based on isozymes-banding pattern, were then analyzed grounded on coefficient correlation between product- moment and goodness of it criteria based on correlation. The results pointed out that morphologically, on eight observed samples which were consist of four different types (species), each Xanthosoma from different locations did not indicate obvious differences. Esterase was formed four different banding-patterns, Glutamate Oxaloacetate Transaminase indicated eight different banding-patterns, and Peroxidase indicated seven different banding-patterns. Correlation between morphological data and data from EST and GOT isozymic banding pattern were very good (0.967918 and 0.937113), While, the correlations between morphological data and POD isozymes were good (0.892721). Key words: kimpul, Xanthosoma, morphological characteristic, isozyme

In Vitro Stability of Phytase from Recombinant Bacteria E. Coli BL21 (DE3) EAS1-AMP

The objective of the research was to inquire the Km, Vm, activity, intracellular phytase stability exposed to pH variation, temperature variation and protease (pepsin and pancreas) in vitro. The phytase was produced from recombinant bacteria E. coli BL21(DE3) EAS1-AMP using 1.5 mM IPTG as inducer. Intracellular enzyme was extracted via freeze shock and centrifugation. Pure enzyme was acquired through NI-NTA agarose column. The enzyme was then tested for Km, Vm, phytase activity and stability against pH, temperature and protease. Treatment levels for stability against protease were P0: without protease, P1: addition of pepsin, P2: addition of pepsin and pancreas, and the data were statistically analyzed using analysis of variance of one-way Completely Randomized Design. Crude intracellular phytase had Vm 6.39 υM/sec, Km 34.82 υM, and 277 units activity. Intracellular phytas was stable at pH 4–6 and 0–550 C. Protease level influenced the activity of intracellular phytase (P<0.05). Intracellular phytase was stable against pepsin but not pancreas. Keywords: Km, Vm, activity, intracellular phytase, pH, temperature, protease

Protein expression on Cr resistant microorganism using electrophoresis method

Abstrak. Fatmawati U, Suranto, Sajidan. 2009. Ekspresi protein pada mikroorganisme resisten Cr dengan metode elektroforesis. Nusantara Bioscience 1: 31-37. Krom heksavalen (Cr(VI)) dikenal sebagai logam berat beracun, sehingga perlu direduksi menjadi Cr(III) yang lebih rendah toksisitasnya. Pseudomonas aeruginosa, Pseudomonas putida, Klebsiella pneumoniae, Pantoea sp. dan Saccharomyces cerevisiae adalah mikroorganisme resisten dan mampu mereduksi Cr(VI). Tujuan penelitian ini adalah mengetahui kemampuan mikroorganisme dalam mengurangi Cr(VI) dan mengetahui pola pita protein antara mikroorganisme resisten Cr(VI) dan mikroorganisme tidak resisten yang diinokulasi pada medium kaldu LB. SDS-PAGE digunakan untuk mengetahui ekspresi protein, sementara konsentrasi Cr(VI) diidentifikasi dengan metode 1,5 difenilkarbazid. Data kuantitatif dianalisis dengan ANAVA dua faktorial dilanjutkan dengan uji jarak berganda Duncan pada taraf 1%. Data kualitatif yaitu ekspresi protein dianalisis dengan mobilitas relatif (Rf). Hasil penelitian menunjukkan bahwa kemampuan mikroorganisme dalam mereduksi Cr(VI) pada konsentrasi awal 0.5 ppm, 1 ppm, 5 ppm dan 10 ppm berbeda-beda, persentase rata-rata kemampuan masing-masing mikroorganisme dalam mereduksi Cr(VI) adalah: P. putida (65%) > S. cerevisiae (64,45%) > P. aeruginosa (60,73%) > Pantoea sp. (50,22%) > K. pneumoniae (47,82%) > tanpa mikroorganisme (34,25%). Penambahan mikroorganisme secara nyata mempengaruhi reduksi Cr(VI). SDS-PAGE menunjukkan bahwa ekspresi protein antara mikroorganisme resisten dan tidak resisten tidak berbeda, tetapi mikroorganisme resisten memiliki lebih banyak protein (pita protein lebih tebal). Kata kunci: logam berat Cr, mikroorganisme, protein, elektroforesis

Bioinformatika: Trend dan Prospek dalam Pengembangan Keilmuan Biologi

PENDAHULUAN Tulisan ini terutama dimaksudkan untuk pengguna awal bioinformatika, termasuk mereka yang selama ini belum mengenal bioinformatika dan tertarik untuk memulainya[1]. Tapi bagi yang selama ini sudah menggeluti bioinformatika secara otodidak melalui internet terutama, cenderung mengalami kesalahan bila tidak memilki dasar-dasar yang kuat terhadap biologi molekuler atau menemui kebuntuan untuk melihat permasalahan biologi yang bisa dipecahkan dengan bioinformatika. Maka dari itu dengan penjelasan agak mendalam tentang prinsip biologi molekuler termasuk bagaimana data biologi molekuler itu didapatkan, diharapkan pembaca makalah ini bisa lebih optimal menggunakan bioinformatika khususnya menunjang pengembangan keilmuan biologi di Tanah Air[2]. [1] Witarto, A.B. Bioinformatika: Mengawinkan teknologi informasi dengan bioteknologi. Trendnya di dunia dan prospeknya di Indonesia. Disampaikan pada Seminar Seminar Teknologi Informasi diselenggarakan oleh MIFTA, Bogor, 9 Januari 2003. Bisa diunduh dari witarto.wordpress.com. [2] Witarto, A.B. Bioinformatics in Indonesia. Disampaikan pada First ASEAN-India Workshop on Bioinformatics di Center for DNA Fingerprinting and Diagnostics, Hyderabad, India, 7-11 November 2005. Bisa diunduh dari witarto.wordpress.co

Pengembangan modul discovery learning struktur tumbuhan dengan memanfaatkan potensi lokal umbul tlatar untuk meningkatkan regulasi diri dan sikap kepedulian lingkungan siswa SMA

Pemanfaatan potensi lokal merupakan salah satu alternatif yang dapat digunakan untuk membantu siswa mempelajari materi biologi. Penggunaan modul DiscoveryLearning (DL) dengan menggunakan potensi lokal Umbul Tlatar diharapkan dapat mempermudah pemahaman siswa terutama pada materi struktur tumbuhan. Tujuan penelitian ini yaitu untuk: 1) mengetahui karakteristik modul DL; 2) mengetahui kelayakan modul DL; 3) mengetahui efektivitas modul DL terhadap keterampilan regulasi diri dan sikap kepedulian lingkungan peserta didik. Metode penelitian menggunakan model penelitian dan pengembangan Borg&amp;Gall yang dimodifikasi yaitu: 1)penelitian pendahuluan dan pengumpulan informasi 2) perencanaan 3)pengembangan desain produk awal 4)uji coba kelompok kecil 5)revisi produk utama 6)uji lapangan terbatas 7)revisi produk akhir. Instrumen yang digunakan meliputi: angket, observasi, wawancara dan tes. Data yang diperoleh yaitu data hasil validasi ahli, validasi praktisi pendidikan. Hasil uji coba dianalisis secara deskriptif kuantitatif. Hasil penelitian dan pengembangan menunjukkan 1) karakteristik modul dikembangkan berdasarkan sintaks Discovery Learning  meliputi: stimulation, problem  statement, data collection, data processing,  verification  dan generalization,  2) Modul dinyatakan layak oleh ahli materi, ahli penyajian modul, ahli perangkat pembelajaran. Modul dinyatakan layak oleh guru dan siswa, 3) Hasil uji operasional terkait regulasi diri menunjukkan signifikansi 0,00&lt;0,05 dan hasil uji operasional  terkait kepedulian lingkungan menunjukkan signifikansi 0,00&lt;0,05, disimpukan modul Discovery Learning(DL) dengan potensi lokal Umbul Tlatar  bermanfaat secara efektif dapat meningkatkan regulasi diri dan sikap kepedulian lingkungan siswa SMA. Development of discovery learning module plant structures by using potential of umbul tlatar area to enhance self  regulation capability and awarness toward  environment attitude of high school students. Utilization of local potential is an alternative that can be used to help students learn biological material. The use of the Discovery Learning (DL) module by using the potential of Umbul Tlatar area is expected to facilitate students' understanding, especially in plant structure materials. The purpose of this research is to: 1) knowing the characteristics of DL modules; 2) knowing the feasibility of the DL module; 3) knowing the effectiveness of the DL module on the ability of self regulation and the Awarness toward environment of students. The research method uses a modified Borg &amp; Gall research and development model, namely: 1) preliminary research and information gathering 2) planning 3) initial product design development 4) small group trials 5) main product revisions 6) limited field tests 7) product revisions. The instruments that used include: questionnaires, observation, interviews and tests. The data obtained are the results of expert validation data, validation of education practitioners, and the results of the trial were analyzed quantitatively descriptively. The results of research and development show 1)module characteristics developed based on discovery learning syntax include: stimulation, problem statement, data collection, data processing, verification, and generalization 2) the module is feasible declared by material experts, module presentation experts, learning device experts.  The module is  declared appropriate by teachers and students 3) the operational test results related to self-regulation show a significance 0.00 &lt;0.05 and the results of operational tests related to environmental care show a significance 0.00 &lt;0.05, it can be concluded that the Discovery Learning module (DL) by using the local potential of useful Umbul Tlatar effectively in improving self-regulation and environmental care attitudes of high school students

Production and Characterization of Crude Intracelluler Phytase From Recombinant Bacteria Peas1amp

This research was aimed at producing a crude intracellular phytase characterized from recombinantbacteria. The recombinant bacteria (pEAS1AMP) was produced by way of transforming pET-22b(+)+pEAS1 into competent E. coli BL21 and E. coli BL21(DE3) cells. Crude intracellular phytaseproduction was induced using 1,5 mM Isopropyl-β-D-thiogalactopyranosid (IPTG). Recombinantbacteria product and enzyme activity test followed the Sajidan method. E. coli BL21(+)pEAS1 and E.coli BL21 (DE3)(+)pEAS1 recombinant bacteria showed growth after 20 hours and 10 hours oftransformation. Phytase activity of E. coli BL21 (DE3)(+)+pEAS1 showed higher than those of E. coliBL21(+)+pEAS1. Crude intracellular phytase of pEAS1AMP recombinant bacteria has an optimumactivity at pH 5, 40oC, incubation period of 60 minutes, substrate concentration of 2%, molecular weight(MW) of 47.3 kDa, Km = 15.91 υM and Vm = 2.41 υM/second. Mg2+ acts as a cofactor but Fe3+ (10-4M) acts as an inhibitor