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Pengaruh Variasi pH dan NaCl terhadap Produksi Inhibitor Protease yang Dihasilkan oleh Acinetobacter baumanii (Bakteri yang Berasosiasi dengan Spons Plakortis nigra) Tati Nurhayati; Maggy T. Suhartono; . Desniar; Rini Suwardinni
Jurnal Pengolahan Hasil Perikanan Indonesia Vol 9 No 2 (2006): Buletin Teknologi Hasil Perikanan
Publisher : Department of Aquatic Product Technology IPB University in collaboration with Masyarakat Pengolahan Hasil Perikanan Indonesia (MPHPI)

Mengingat semakin jelasnya keterlibatan protease dalam penyebab penyakit, maka diperlukan suatu senyawa untuk menghambat aktivitas enzim tersebut, yaitu inhibitor protease. Senyawa tersebut dapat dihasilkan oleh makro dan mikroorganisme laut. Untuk memproduksi inhibitor protease diperlukan suatu kondisi produksi yang tepat, termasuk NaCl dan pH media. Oleh karena itu tujuan penelitian ini adalah menentukan konsentrasi NaCl dan pH yang tepat dalam media untuk menghasilkan inhibitor protease dengan aktivitas tertinggi. Metode yang digunakan untuk mencapai tujuan tersebut adalah dengan menumbuhkan Acinetobacter baumanii dalam media marine broth+glukosa 0,05 %(w/v) dengan melakukan variasi NaCl (0 %,2 %, dan 4 %(w/v)) dan juga pH (6,7, dan 8). Berdasarkan hasil penelitian didapatkan bahwa konsentrasi NaCl yang tepat untuk produksi maksimum adalah 2 %(w/v) dan pH media 8 dengan waktu inkubasi 20-28 jam. Aktivitas tertinggi yang dihasilkan adalah 1,64-1,93 U/ml.Kata kunci: Acinetobacter baumanii, inhibitor protease

Modifikasi Media Marine Broth pada Produksi Inhibitor Protease dari Bakteri Acinetobacter baumanni yang Hidup Bersimbiosis dengan Sponge Plakortis nigra . Desniar; Tati Nurhayati; Maggy T. Suhartono; Eko Muhammad Isa
Jurnal Pengolahan Hasil Perikanan Indonesia Vol 9 No 1 (2006): Buletin Teknologi Hasil Perikanan
Publisher : Department of Aquatic Product Technology IPB University in collaboration with Masyarakat Pengolahan Hasil Perikanan Indonesia (MPHPI)

Enzim protease memiliki peranan penting dalam proses metabolisme bakteri patogen. Protease bakteri tersebut dapat dihambat aktivitasnya oleh inhibitor protease. Iinhibitor protease dapat diproduksi dari bakteri Acinetobacter baumanni yang hidup bersimbiosis dengan sponge laut (Plakortis nigra). Agar produksi inhibitor protease dapat lebih optimal maka perlu diketahui komposisi media marine broth yang baik. Perlakuan yang digunakan pada penelitian ini adalah modifikasi konsentrasi peptone (0,5% dan 1%) dan modifikasi konsentrasi yeast extract (0,1%; 0,5% dan 1%). Peningkatan konsentrasi yeast extract memperpanjang waktu propagasi bakteri A. baumanni dan memperlambat awal produksi inhibitor. Peningkatan konsentrasi pepton cenderung menghasilkan pertumbuhan yang lebih baik. Aktivitas penghambatan inhibitor protease tertinggi terjadi pada media dengan kombinasi konsentrasi pepton dan yeast extract masing-masing 0,5%.Kata kunci: inhibitor protease, Acinetobacter, simbiosis sponge, produksi

Karakteristik Protease dari Bakteri Patogen Staphylococcus epidermidis Ace Baehaki; Tati Nurhayati; Maggy T. Suhartono
Jurnal Pengolahan Hasil Perikanan Indonesia Vol 8 No 2 (2005): Buletin Teknologi Hasil Perikanan
Publisher : Department of Aquatic Product Technology IPB University in collaboration with Masyarakat Pengolahan Hasil Perikanan Indonesia (MPHPI)

Beberapa bakteri patogen memproduksi enzim hidrolitik seperti protease dan hialuronidase yang berfungsi untuk mendegradasi komponen matrik ekstraseluler sehingga dapat merusak struktur jaringan inang. Tujuan penelitian ini adalah untuk melakukan karakterisasi protease dari bakteri Staphylococcus epidermidis yang diisolasi dari koleksi Rumah Sakit Pertamina Jakarta. Bakteri ditumbuhkan pada media Luria broth (LB) yang mengandung tryptone 1 %, NaCl 1 % dan yeast extract 0,5 %. Hasil karakterisasi menunjukkan bahwa protease S. epidermidis ini memiliki pH dan suhu optimum 8 dan 50 oC. Ion logam Mn2+ (5 mM) dan Ba2+ (5 mM) merupakan aktivator kuat protease S. epidermidis yang dapat meningkatkan aktivitas protease masing-masing 3 dan 2 kali lipat dari protease kontrol, sedangkan Na+ (1 mM), K+ (1 mM), Fe3+ (1 dan 5 mM), Zn2+ (5 mM), dan Ca2+ (1 mM) merupakan inhibitor ion logam yang kuat. Protease S. epidermidis digolongkan ke dalam serin metaloprotease karena dapat dihambat secara sempurna oleh PMSF dan EDTA. Protease tersebut mempunyai berat molekul sekitar 35 kD.Kata Kunci: bakteri patogen, karakterisasi, protease, Staphylococcus epidermidis.

Penapisan Inhibitor Protease yang Dihasilkan oleh Sponge Asal Kepulaun Seribu Tati Nurhayati; Pipih Suptijah; Maggy T. Suhartono; Irman Febrian
Jurnal Pengolahan Hasil Perikanan Indonesia Vol 7 No 2 (2004): Buletin Teknologi Hasil Perikanan
Publisher : Department of Aquatic Product Technology IPB University in collaboration with Masyarakat Pengolahan Hasil Perikanan Indonesia (MPHPI)

Several medicines showing protrease inhibitor mechanism are widely available in the markets. Among marine organisms, sponge is the big producer of bioactive compound, include protease inhibitor. The purpose of this research was to screen of protease inhibitor produced by sponge. Screening was be conducted using agar diffusion methods on ten of sponge which the extracted with methanol and distilled water. As test bacteria were used three species of pathogenic bacteria that is Escherichia coli, Pseudomonas aeruginosa, and Staphyloccocus aureus. The result of research showing that sponge which be extracted using methanol have low protease inhibitory activity (≤ 30%), while that which be extracted using distilled water have high protease inhibitory, that is extract of Jaspis stellifera and extract of Plakortis nigra (≥ 50%). The extract of Jaspis stellifera was inhibited of protease from Escherichia coli and Staphyloccocus aureus by minimum inhibitory concentration (MIC) 0.08%, while that extract of Plakortis nigra was inhibited protease from S. aureus by MIC 0.12%. Ethymenediamine tetraacetic acid (EDTA) was inhibited S. aureus and E. coli by MIC 0.16%. Based on the data, can be concluded that both the extract of sponge were potential as protease inhibitor.Keywords: Bacteria, protease inhibitor, screening, sponge

Karakterisasi Protease dari Bakteri Aeromonas hydrophila Ace Baehaki; Tati Nurhayati; Maggy T. Suhartono
Jurnal Pengolahan Hasil Perikanan Indonesia Vol 7 No 2 (2004): Buletin Teknologi Hasil Perikanan
Publisher : Department of Aquatic Product Technology IPB University in collaboration with Masyarakat Pengolahan Hasil Perikanan Indonesia (MPHPI)

In the last decade, a concern on protease as medicinal target for overcoming bacterial and viral diseases has been rapidly increased because of the obvios involvement of this enzyme in the molecular of the diseases mechanism. The porpuse of this research was to characterize proteases from fish pathogenic bacteria Aeromonas hydrophila. The bacteria were grown in media containing triptone 1%, NaCl 1% and yeast extract 0,5%. The optimum production time of A. hydrophila was 48 h, the optimum pH was 7,5, the optimum temperature was 50oC. Study on the effect of metals ion and spesific inhibitors indicated that protease from A. hydrophila was serin metaloprotease.Keywords: protease, characterization, pathogenic bacteria

Penapisan Senyawa Bioaktif pada Siput Laut Gonggong (Laevistrombus turturella) Asal Bintan : Bioactive Compound Screening of Gonggong Snail (Laevistrombus turturella) Lily Viruly; Nuri Andarwulan; Maggy T. Suhartono; Mala Nurilmala
Jurnal Pengolahan Hasil Perikanan Indonesia Vol 23 No 2 (2020): Jurnal Pengolahan Hasil Perikanan Indonesia 23(2)
Publisher : Masyarakat Pengolahan Hasil Perikanan Indonesia (MPHPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.17844/jphpi.v23i2.32041

Siput laut gonggong asal Bintan merupakan salah satu gastropoda laut yang belum dimanfaatkan secara optimal. Siput ini merupakan makanan laut khas Bintan dan harganya sangat mahal. Secara empiris, siput ini dipercaya dapat meningkatkan stamina dan vitalitas. Tujuan penelitian ini adalah untuk melakukan penapisan senyawa bioaktif pada siput laut gonggong asal Bintan yaitu aktivitas antioksidan dan aktivitas antimikroba. Aktivitas antioksidan dianalisis menggunakan metode DPPH dan aktivitas antimikroba dianalisis menggunakan metode difusi sumur. Aktivitas antioksidan ekstrak gonggong bercangkang tipis dengan IC50 1.433,08±0,01 ppm lebih tinggi daripada gonggong bercangkang tebal dengan IC50= 2.051,55±0,10 ppm. Aktivitas antimikroba pada ekstrak gonggong lebih baik pada bakteri Gram positif daripada bakteri Gram negatif. Ekstrak gonggong rebus bercangkang tebal memiliki aktivitas antimikroba yang paling tinggi dengan nilai rata-rata diameter daya hambat (DDH) yaitu 25,53±0,12 mm.

Silica, a polimerized silicon dioxide, is widely used as raw materials for food industries, such as food packaging, filter agent, biomarkers and biosensor for various analysis. In biological sistem such as sponge, the formation of silica structure was directed by protein known as silicatein. The aims of this research were to extract silicatein-like protein isolated from sponge live surrounding the Nias and Lombok seacost Indonesia and to study their activity to polymerize tetraethoxyorthosilic M.R.R. Lukie Trianawati; Maggy T. Suhartono; Dahrul Syah; Ekowati Chasanah
Forum Pasca Sarjana Vol. 31 No. 2 (2008): Forum Pascasarjana
Publisher : Forum Pasca Sarjana

Show Abstract | Download Original | Original Source | Check in Google Scholar

Silica, a polimerized silicon dioxide, is widely used as raw materials for food industries, such as food packaging, filter agent, biomarkers and biosensor for various analysis. In biological sistem such as sponge, the formation of silica structure was directed by protein known as silicatein. The aims of this research were to extract silicatein-like protein isolated from sponge live surrounding the Nias and Lombok seacost Indonesia and to study their activity to polymerize tetraethoxyorthosilicate (TEOS) in-vitro. Protein in silica spicule was isolated by collecting silica spicule, soaked in HF/NH4F buffer (pH5.0) for dissolving silica and releasing this protein. The protein was analysed by electrophoresis SDS-PAGE to estimate the molecular weight and the concentration was analyzed by Bradford method. The highest yield of silica spicula was 58.5% of dry weight sponge that was isolated from sponge MT37. By SDS-PAGE, the molecular weight of protein from N6 showed three bands of 32, 27, 23 kDa, while MT5 protein was 15.5 kDa, and MT37 protein was 18 kDa. The highest polymerization activity was 144 µmol/ml TEOS occurs at 12 hours, showed by protein isolated from sponge MT37 of Lombok Marine. Key words: sponge, silicatein like-protein, tetraethoxyorthosilicate

Pemurnian dan Karakterisasi Kitosanase Bacillus coagulans LH 28.38 Winda Haliza; Maggy T. Suhartono; Dahrul Syah
Jurnal Penelitian Pascapanen Pertanian Vol 2, No 2 (2005): Jurnal Penelitian Pascapanen Pertanian
Publisher : Balai Besar Penelitian dan Pengembangan Pascapanen Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jpasca.v2n2.2005.76-83

Enzim ekstrasellular kitosanase telah berhasil dimurnikan dan dikarakterisasi dari bakteri termofilik Bacillus coagulans LH 28.38 asal tanah geotermal Lahendong - Sulawesi Utara. Media pertumbuhan mengandung 0,4% koloidal kitosan sebagai sumber karbon dan diproduksi optimum pada suhu 55°C selama tujuh hari. Pemurnian enzim dilakukan dengan teknik filtrasi gel menggunakan matriks Sephadex G-IOO menghasilkan tiga puncak protein sebagai kitosanase fraksi A. B, C dan masing masing memiliki aktivitas sebesar 13.33 U/mg, 38.66 U/mg dan 88 U/111g. Kitosanase fraksi A memiliki suhu optimum pad a 60°C dan bekerja pad a kisaran pH luas 2-12. Kitosanase fraksi B memiliki suhu optimum 70°C dan bekerja pada kisaran pH 2-12. Kitosanase f'raksi C memiliki suhu optimum 60°C dan kisaran pH pH 2-9. Penentuan berat molekul enzim dilakukan dengan cara SDS-PAGE dan anal isis zimogram. Berm molekul enzim yang diperoleh 93, 91, 87, 77.6 dan 74 kDa. Enzim kitosanase ini dapat diaplikasikan untuk memperoleh oligomer kitosan. Oligomer kitosan sangat diminati oleh kalangan medis, karena mamiliki aktivitas anti tumor, anti kolesterol dan anti mikroba. Purification lind characterization of chitosanase from Bacillus coagulans LH 28.38Characterization and purification of the extracellular chitosanase produced by Bacillus coagulans LH 28.38 from geothermal soil origin from Lahendong - North Sulawesi has been successfully done. The enzyme was produced in medium containing 0.4% colloidal chitosan as sole carbon source and reached the highest level after 7"' day incubation at 55°C. The purification of enzyme was done by column chromatography using Sephadex G-IOO as matrix produced three peaks of protein chitosanase named as Chitosanase fraction A, Band C while has 13.3 U/mg; 38.66 U/mg and 88 U/mg activity, respectively. Chitosanase fraction A has optimal temperature at 60°C and work at range pH 2-12. Chitosanase fraction B has optimal temperature at 70°C and work at range pH 2-12. Chitosanase fraction C has optimal temperature 70°C and work at range pH 2-9. Molecular weight of chitosanase was determined by SDS-PAGE and zymograrn analysis. The molecular weight of enzyme was estimated to be 93, 91. 87. 77.6 and 74 kDa. This chitosanase could be applied to produce oligochitooligosaccaride. Oligochitooligosaccaride was used for medical popuse by medical, due to have the hipokolesterolemik, antimicrobial and antitumor activities.

Pemurnian Ekstraseluler Hyaluronidase Streptococcus agalactiae (Streptokokus Grup B ) (Extracellular Hyaluronidase Purification of Streptococcus agalactiae (Group B of Streptococus) Wendry Setiyadi Putranto; Sri Budiarti; Maggy T. Suhartono; I Wayan T. Wibawan; Zainatul Hayati
Jurnal Ilmu Ternak Vol 6, No 1 (2006)
Publisher : Fakultas Peternakan, Universitas Padjadjaran

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24198/jit.v6i1.2260

Hyaluronidase (EC 4.2.2.1) merupakan enzim ekstraseluler yang dihasilkan Streptococcus agalactiae (Streptokokus Grup B). Penelitian ini bertujuan untuk mengetahui aktivitas spesifik dari hyaluronidase S. agalactiae dan berat molekul proteinnya. Pemurnian enzim dengan sentrifugasi, pengendapan amonium sulfat 45% dan kromatographi kolom dengan Sephadex G-100 dan Sodiumdodecyl sulfate polyacrylainide gel electrophoresis (SDS PAGE). Enzim hyaluronidase dari S. Agalactiae memiliki aktivitas spesifik sebesar 0,0012 U/mg (ekstrak kasar), 0,0125 U/mg (pengendapan dengan 45% ammonium sulfate) and 0,032 U/mg (Gel Filtration). Berat molekul protein hyaluronidase adalah 102 - 106 kD.Kata kunci : hyaluronidase, aktivitas spesifik, pemurnian

Aktivitas IgY dan IgG Antitetanus setelah Perlakuan pada Berbagai pH, Suhu dan Enzim Proteolitik I Gusti Ayu Agung Suartini; I Wayan Teguh Wibawan; Maggy T. Suhartono; Supar -; I Nyoman Suarta
Jurnal Veteriner Vol 8 No 4 (2007)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (259.61 KB)

A study was carried out to find out an alternative method of producing antitetanus antibody (IgY) in chicken and to evaluate its activity at different levels of pH, temperatures and proteolytic enzymes. Antitetanus IgY was produced by immunization of chickens with tetanus toxoid, three times weekly at gradual doses of 100, 200, and 300 Lf, respectively. Serum samples were collected 4 weeks following the last immunization. IgY was purified by ammonium sulfat precipitation and gel filtration chromatography (Sephadex G. 120).The purified IgY was then treated at different levels of temperatures and pH as well as proteolytic enzymes. Commercial antitetanus IgG was used as control. The activities of treated IgY and IgG were tested by enzyme linked immunosorbent assay (ELISA). IgY and IgG activities were significantly reduced at 80ºC and completely destroyed at 90ºC. Treatment with pepsin significantly reduced IgY and IgG whereas trypsin slightly reduced IgY activities and has no effect on IgG activities. IgY and IgG activities were reduced significantly at pH < 3 and and only sightly reduced at pH>10. It is evident that heating at >90oC, pH at <3 and treatment with pepsin significantly reduced IgY activities and it appears that IgG was more resistent to the efect of temperatures, pH and proteolytic enzymes

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