Articles

Preliminary Characterization of Protease Inhibitor from Bacteria-Associated with Sponge from Panggang Island, Seribu Islands NURHAYATI, TATI; SUHARTONO, MAGGY THENAWIDJAJA; NURAIDA, LILIS; POERWANTO, SRI BUDIARTI
HAYATI Journal of Biosciences Vol 13, No 2 (2006): June 2006
Publisher : Bogor Agricultural University, Indonesia

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Abstract

Pathogenic bacteria produced protease that involved in molecular mechanism of foodborne disease. Produced protease involved in molecular mechanisms of foodborne diseases. The purpose of this research was to screen, identify and characterize the potential microorganisms associated with sponge as producer of protease inhibitor. Among 96 isolates examined, four isolates i.e 10A6, 6A3, 9A51, and 1A12 yielded protease inhibitors which were potential to inhibit protease substrates (40-90%). One of the most potential protease inhibitor producer, the bacteria isolate 6A3, was identified as Chromohalobacter sp. Chromohalobacter sp.6A3 produced protease inhibitor with optimum temperature and pH 300 C and 5, respectively. The inhibitor activity was stable when incubated at 400 C for ten minutes or at 300 C for 8 hours. Key words: Bacteria, Chromohalobacter sp., protease inhibitor, screen, sponge
Purification and Charaterization of Protease from Pathogenic Bacteria Pseudomonas aeruginosa Baekhari, Ace; Suhartono, Maggy T; Palupi, Nurheni Sri; Nurhayati, Tati
Jurnal Teknologi Dan Industri Pangan Vol 19, No 1 (2008): Jurnal Teknologi dan Industri Pangan
Publisher : Departemen Ilmu dan Teknologi Pangan, IPB Indonesia bekerjasama dengan PATPI

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Abstract

In the last decade, concern on protease as medical target for overcoming bacterial diseases and viral diseases has been rapidly increased because of the obvious involvement of this enzyme in the molecular of the diseases. The purpose of this research was to purify and characterize protease from pathogenic bacteria Pseudomonas aeruginosa. The bacteria were grown in media containing triptone 1%, NaCl 1% and Yeast extract 0.5%. protease of Pseudomonas aeruginosa was purified using column chromatography with Sephadex G-100 gel. There were three peaks of enzyme protein, which were detected on fractions 14,17 and 30. the optimum pH of the extracelluler protease from Pseudomonas aeruginosa was 8. The optimum temperature of Pseudomonas aeruginosa was 300C. Fe3+ (1dan 5 mM) was strong activator and Co 2+ was strong inhibitor. Study on the effect of metals ion and specific inhibitors indicated that protease from Pseudomonas aeruginosa was serin metaloprotease. The apparent moleculer weights, as determined by SDS-PAGE and zymogram technique, 36 kD and 42 kD. Key word : Protease, characterization, purification, pathogenic bacteria P. aeruginosa
Optimasi dan pemodelan proses recover flavor dari limbah cair industri pengolahan rajungan dengan reverse osmosis Uju, Uju; Ibrahim, Bustami; Trilaksani, Wini; Nurhayati, Tati
Jurnal Ilmu Pertanian Indonesia Vol 14, No 1 (2009): Jurnal Ilmu Pertanian Indonesia
Publisher : Institut Pertanian Bogor

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Abstract

The waste water of blue crab pasteurization has potential in environmental pollution. It contained TSS of 206.5mg 1-1, BOD 7,092.6mg.1-1 and COD of 51,000mg.1-1 on the other hand, it also contains an interesting flavor compound, which composed of 0.23°/o non protein nitrogen and 17 amino acids where the highest was glutamic acid one. In this study, pre-filtration step using filter size 0.3 1J followed by reverse osmosis has been used to reduce these pollutions load and flavor compound recovery. During pre-filtration steps, TSS was reduced to 74.8°/o so turbidity decrased reased until 31°/o. After reverse osmosis process, BOD, and COD decreased more than 99°/o, and there was no amino acids detected in permeate stream. Factors that affect performance of reverse osmosis were transmembrane pressure, temperature and pH. The higher transmembrane pressure, temperature and pH resulted the higher the flux permeate. The use of higher temperature make flux increasing, eventually increasing transmembrane pressure make the flux increased only at transmembrane pressure less than 716 kPa. The protein rejection was influenced unsignifanctly by transmembrane pressure, temperature and pH. During concentrating flux declined exponentially by time function. At concentration factor 2.75 resulted 79°/o and 12°/o of increasing protein and NPN, respectively. The amino acids content can be increased 2-23 times of the origin. Even arginin and sistin, the amino acids that were undetectable initially, but they can bedetected at concentration of 0.0360 and 0.0250 (w/v) respectively at the end of the process. Hidrolysis and fermentation process can increase the amino acid content 31-45 times. Keywords : Blue crab, flavor, recovery, reverse osmosis.
PEMURNIAN DAN KARAKTERISASI INHIBITOR PROTEASE DARI Chromohalobacter sp. 6A3, BAKTERI YANG BERASOSIASI DENGAN SPONS Xetospongia testudinaria [Purification and Characterization of Protease Inhibitor from Chromohalobacter sp. 6A3, Bacteria-associated with S Nurhayati, Tati; Suhartono, Maggy T.; Nuraida, Lilis; Poerwanto, Sri Budiarti
Jurnal Teknologi Dan Industri Pangan Vol 21, No 2 (2010): Jurnal Teknologi dan Industri Pangan
Publisher : Departemen Ilmu dan Teknologi Pangan, IPB Indonesia bekerjasama dengan PATPI

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Abstract

PEMURNIAN DAN KARAKTERISASI INHIBITOR PROTEASE DARI Chromohalobacter sp. 6A3, BAKTERI YANG BERASOSIASI DENGAN SPONS Xetospongia testudinaria [Purification and Characterization of Protease Inhibitor from Chromohalobacter sp. 6A3, Bacteria-associated with Sponge Xetospongia testudinaria]           Tati Nurhayati1)*, Maggy T. Suhartono2), Lilis Nuraida2), Sri Budiarti Poerwanto3) 1) Departemen Teknologi Hasil Perairan, Fakultas Perikanan dan Ilmu Kelautan, Institut Pertanian Bogor 2) Departemen Ilmu dan Teknologi Pangan, Fakultas Teknologi Pertanian, Institut Pertanian Bogor 3) Departemen Biologi, Fakultas Matematika dan Ilmu Pengetahuan Alam, Institut Pertanian Bogor   Diterima 26 November 2009/ Disetujui  15 Desember 2010 ABSTRACT   Various sponges has been reported to produce protease inhibitor which could inhibit protease activity of pathogenic bacteria. The previous research showed that bacteria-associated with sponge could produce bioactive compound similar to their host, including protease inhibitor. The purposes of this research were to purify protease inhibitor from Chromohalobacter sp. 6A3 and to study the characteristics of the protease inhibitor. The result showed that the protein can be extracted by 30 % (v/v) acetone, purified by gel filtration (Sephadex G-75) and finally, purified by anion exchanger (Sephadex A-50). The molecular weight of the purified protease inhibitor after gel filtration was estimated as 21,31 kDa and 17,05 kDa, but anion exchanger gave protein with estimated molecular weight of 21,31 kDa The optimum temperature and pH were 30 oC and 5 respectively. The protease inhibitor could resist heating at 30 oC for 10 minutes. Incubation of the inhibitor at 30 oC, pH 5, still retained its activity until 3 hours. The purified enzyme inhibitor was also still active after incubated at pH from 5 to 6 for 1 hour. The most susceptible substrate (enzyme) for the inhibitor was protease from P. aeruginosa. The protease inhibitor was inhibited by metal ions except Na+ 1mM. Activity of the inhibitor increased twofold by SDS 5 mM. IC 50 of the protease inhibitor was 3.48 nM. The protease inhibitor inhibited the enzyme uncompetitively.   Key words: chromohalobacter, protease inhibitor, sponge  
Aktivitas Inhibitor Protease dari Ekstrak Karang Lunak, Asal Perairan Pulau Panggang Kepulauan Seribu Nurhayati, Tati; Fikri, Muhammad; Desniar, Desniar
ILMU KELAUTAN: Indonesian Journal of Marine Sciences Vol 15, No 2 (2010): Jurnal Ilmu Kelautan
Publisher : Marine Science Department Diponegoro University

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Abstract

Beberapa komponen bioaktif dihasilkan oleh karang lunak, salah satunya inhibitor protease. Tujuan penelitian ini adalah mendapatkan karang lunak yang berpotensi sebagai penghambat aktivitas kerja enzim protease (inhibitor protease) pada beberapa bakteri patogen penghasil enzim protease serta mengetahui Minimum Inhibitory Concentration (MIC) dari ekstrak karang lunak tersebut.  Hasil penelitian menunjukkan bahwa pelarut metanol  lebih  potensial  untuk  mengekstrak  inhibitor  protease  dari  karang  lunak.  Karang  lunak  jenis Sarcophyton sp. dan Sinularia sp. mampu menghambat 100% aktivitas protease bakteri Staphylococus aureus dengan MIC 0,04% lebih kecil dari pada MIC EDTA (0,16%), sedangkan Xenia sp. menghambat protease bakteri S. aureus dengan MIC 0,08%. Karang lunak Nephthea sp. menghambat protease bakteri Pseudomonas aeruginosa dengan MIC 0,28%. Kata kunci : inhibitor protease, karang lunak, MIC Several bioactive compounds were produced by soft corals, including protease inhibitor.The aim of this study was to obtain softcorals which potency as inhibitor toward protease enzyme activity on pathogenic bacterial that produced protease enzyme and to study Minimum Inhibitory Concentration (MIC) from the softcorals. This research shown that ethanol is more potential for extracting protease inhibitor from softcorals. Sarcophyton sp. and Sinularia sp. are capable of inhibiting protease enzyme activity against Staphylococus aureus as 100% by MIC 0.04%, while that EDTA had MIC toward the protease as 0.16%.  Xenia sp. was capable of inhibiting protease from S. aureus by MIC 0.08%.  In the otherhand Nephtea sp. inhibited protease from Pseudomonas aeruginosa by MIC 0.28%. Key words: protease inhibitor, soft coral, MIC.
PEMBUATAN DAN KARAKTERISASI HIDROLISAT PROTEIN DARI IKAN LELE DUMBO (Clarias gariepinus) MENGGUNAKAN ENZIM PAPAIN Salamah, Ella; Nurhayati, Tati; Widadi, Indah Rahayu
Jurnal Pengolahan Hasil Perikanan Indonesia Vol 15, No 1 (2012): Jurnal Pengolahan Hasil Perikanan Indonesia
Publisher : Departement of Aquatic Product Technology

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Abstract

African catfish (Clarias gariepinus) is important freshwater fish commodity in Indonesia. African catfish also has high protein content, which is potential to be used as protein hydrolysate. Fish protein hydrolysate offered many advantages in food, feed, agriculture, microbiology and medicinal’s field. This research aimed to determine the optimal condition for hydrolysis of African catfish protein and to characterize protein hydrolysate from African catfish. The optimal concentration of papain used for hydrolysis of African catfish protein was 5% (w/v) for 6 hours hydrolysis time and degree of hydrolysis was 35.37%. Protein hydrolysate from African catfish revealed 21.16% of yield and water content of 5.46%; ash content of 5.71%; protein content of 53.29%; and fat content of 1.94%. Protein hydrolysate from African catfish contained 15 types of amino acids, with 9 types of essential amino acids and 6 types of non essential amino acids. The highest essential amino acid composition was lysin of 5.23%, while the highest non essential amino acid composition was glutamic acid of 7.77%. Protein hydrolysate from African catfish showed protein digestibility by in vitro enzymatic hydrolysis of 98.57%.Key words: African catfish, amino acid, fish protein hydrolysate, papain
PURIFIKASI PARSIAL DAN KARAKTERISASI ENZIM KATEPSIN DARI IKAN BANDENG (Chanos Chanos Forskall) Nurhayati, Tati; Salamah, Ella; Dynnar, nico
Jurnal Pengolahan Hasil Perikanan Indonesia Vol 15, No 2 (2012): Jurnal Pengolahan Hasil Perikanan Indonesia
Publisher : Departement of Aquatic Product Technology

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Abstract

Decomposition of protein in the enzymatic process will lead to changes in odor, texture, and appearance of fish. The enzymes that play a role in the enzymatic process is primarily proteolytic enzymes. Cathepsin enzyme is one of the proteolytic enzymes found in animal tissue that hydrolyzes peptide bonds of proteins. Information on optimal conditions of cathepsin enzyme activity is useful in the process of good handling,  to avoid environmental conditions that can increase cathepsin enzymes activity, especially in milkfish. The purposes of this study were to partial purify the catepsin  enzyme of milkfish and characterize the enzymes. Crude extract of enzyme had specific activity 0.8598 U/mg and after presipitation with ammonium sulphate 70% obtained specific activity of 4.4643 U/mg and after 6 hours of dialysis obtained specific activity 14.4404 U/mg. The enzyme cathepsin worked optimally at 40 °C and pH 4, and 3%  substrate. Divalent metal ions inhibited the activity of the enzyme, higher compared to monovalent or trivalent metals. Cathepsin enzyme identified had molecular weight of 86.67 kDa.Key words: cathepsin, characterization, milkfish, purification
Modifikasi Media Marine Broth pada Produksi Inhibitor Protease dari Bakteri Acinetobacter baumanni yang Hidup Bersimbiosis dengan Sponge Plakortis nigra Desniar, .; Nurhayati, Tati; Suhartono, Maggy T.; Isa, Eko Muhammad
Jurnal Pengolahan Hasil Perikanan Indonesia Vol 9, No 1 (2006): Buletin Teknologi Hasil Perikanan
Publisher : Departement of Aquatic Product Technology

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Abstract

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
Characterization and Recovery of Minced fish Wastewater Protein using Reverse Osmosis Uju, .; Nurhayati, Tati; Ibrahim, Bustami; Trilaksani, Wini; Siburian, Maglory
Jurnal Pengolahan Hasil Perikanan Indonesia Vol 12, No 2 (2009): Jurnal Pengolahan Hasil Perikanan Indonesia
Publisher : Departement of Aquatic Product Technology

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Abstract

Recovery of minced fish wastewater soluble protein was done by reverse osmosis membrane. Themincefish wastewater has high level polluting load. It contains COD and TSS 4.233 and 55,06 mg/lrespectively, but it is potential to utilized because of high protein and amino acids content especiallyglutamic acid. Fluxes of permeate during recovery process were kept on steady state in the beginningprocess at 15 l/m2h without prefiltration and 20 l/m2h with 0,9 micron prefiltration. During concentration,flux time decreased from 22,11 l/m2h at concentration factor 1 to 12,95 l/m2h at concentration factor 1,52.In other sides the rejection value of protein increased from 90,11% in the beginning process to 99,50% inthe end of the concentration process. At the end of the recovery stage, yield concentrate contained 0.401 g/lwhich is 21 fold greater than the original step. Therefore permeate stream delivered high quality effluentand met the standard of wastewater effluent.Keywords: minced fish, protein, recovery, reverse osmosis
Karakterisasi Protease dari Bakteri Aeromonas hydrophila Baehaki, Ace; Nurhayati, Tati; Suhartono, Maggy T.
Jurnal Pengolahan Hasil Perikanan Indonesia Vol 7, No 2 (2004): Buletin Teknologi Hasil Perikanan
Publisher : Departement of Aquatic Product Technology

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Abstract

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
Co-Authors . Desniar . Uju A. Zaenal Mustopa Ace Baehaki Ace Baekhari Agoes M Jacoeb, Agoes M Agoes Mardiono Jacoeb Alfi Hamdan Zamzami AMALIA, ELIN Andriyani, Pitria Aprilia, Bintang Efrata Aris Munandar Ary Apriland Aulia Andhikawati Azizah Nuraini Bagus Sediadi Bandol Utomo, Bagus Sediadi Bandol Barokah, Giri Rohmad Bunga, Selvanda Moreine Bustami Ibrahim Desniar Desniar Djailani, Fernandy Eko Muhammad Isa Ekowati Chasanah Ella Salamah Febrina Olivia Akerina, Febrina Olivia Fitriany Podunge, Fitriany Fransiskayana, Andika Gadi, Dewi Setiyowati Hani Novanti, Hani Ifah Munifah Ika Astiana Indah Rahayu Widadi Iriani Setyaningsih Irman Febrian Jacoeb, Agoes Komariah Tampubolon Kustiariyah Tarman LAKSMI AMBARSARI Laksono, Untung Trimo LILIS NURAIDA Made Suhandana Maggy T Suhartono Maggy T. Suhartono Maggy Thenawidjaja MAGGY THENAWIDJAJA SUHARTONO Maglory Siburian Mala Nurilmala Manteu, Shindy Hamidah masdudi masdudi, masdudi Merdekawati, Dewi Mohammad Irfan Mohammad Zaenuri, Mohammad Muhammad Fikri Muhammad Zakiyul Fikri nico Dynnar Ninik Purbosari Nugroho, Teguh Setyo Nurfajrin Nisa, Nurfajrin Nurheni Sri Palupi Nurhikma, Nurhikma Nurjanah . Nurjanah Nurjanah Nurlisa Butet, Nurlisa Nur’aenah, Nani Pasaribu, Ewi Pipih Suptijah Pratitis, Asri Putalan, Reinal Rahmawati, Mutiara Rina Rina Rini Suwardinni Roni Nugraha Roskananda, Rieska Ruddy Suwandi S Nurul Janah, S Nurul Saeful Bahri Santhy Wisuda Sidauruk Saputra, Dede Sri Budiarti Poerwanto Sri Purwaningsih Sugeng Heri Suseno Syefri Rusyadi, Syefri Taufik Hidayat Trilaksani, Win Uju Uju Uzainah Aremhas Wini Trilaksani Yenni Yenni Yulia Oktavia Yuniarti, Tatty Yusro Nuri Fawzya ZAKARIA, RIJAN