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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

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (263.842 KB) | DOI: 10.6066/366

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
Karakteristik Endapan Cairan Rumen Sapi asal Rumah Potong Hewan sebagai Feed Supplement Budiansyah, Agus; Resmi, Resmi; Nahrowi, Nahrowi; Wiryawan, Komang G; Suhartono, Maggy T; Widyastuti, Yantyati
Jurnal Ilmu-Ilmu Peternakan JIIP Volume XIV No. 1 Mei 2011
Publisher : Jurnal Ilmu-Ilmu Peternakan

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (71.575 KB)

Abstract

The  aims  of  this  experiment  were  to    identify  and  characterize of sediment  product from cattle rumen liquor as a source of amino acids, minerals and vitamins. The sediment were obtained as pellets upon centrifugation of rumen liquor at 10,000 g for 10 minutes for collection of supernatant. The sediment were evaluated for amino  acids, minerals and vitamins composition and chararacterized for the pH,  solubility of dry  matter,  specific  density,  bulk  and  compacted  bulk  densities  and  angle  of  response.  Result  of  the experiment showed that sediment contained higher minerals:  Na, K and Fe compared with the commercial premix, but  lower in B-vitamins and  amino acids. The composition of mineral Na,  K, and Fe from rumen liquor of local cattle was 13.47%, 7.73 % and 14.52 %,  while Na, K, and Fe from  rumen liquor of  imported cattle was 18.40%, 10.25%, and 14.52% respectively. The sediment had pH range from 10.01-10.03, the dry matter solubility   was 35.5%  up to 39.1%. The  sediment from imported cattle had  higher specific density, bulk  and  compacted  bulk  densities  and  angle  of  response  than  that  of  local  cattle. It  is  concluded  that sediment from cattle rumen liquor contained high  Na, K  and Fe, low amino acids and B-vitamin,  high pH and low solubility.
Characterization of Phenol Degrading Bacteria from Buntal Lake of Central Kalimantan ., Nursaadah; Santosa, Dwi Andreas; Suhartono, Maggy T
Jurnal Ilmu Tanah & Lingkungan Vol 3, No 2 (2000): Jurnal Ilmu Tanah dan Lingkungan
Publisher : Jurnal Ilmu Tanah & Lingkungan

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (71.575 KB)

Abstract

Three bacterial isolates (ICBB 1168, ICBB 1169, ICBB 1170), being capable to utilizing phenol as sole carbon and energy source, were isolated from Buntal Lake of Central Kalimantan. Growth of all isolates were optimum at 37OC. Optimum pH for growth and phenol degradation of ICBB 1168, lCBB 1169, and ICBB 1170 were 7-8, 6, and 6-7, respectively. Among the isolates, ICBB 1170 showed best phenol degrading activity. ICBB 1170 able to degrade 16 mM phenol to 0.71 mM in 4days. Phenol degradation ability of ICBB 1170 could be increased by adding 0.01-0.1% yeast extract. Addition of 0.05-0.1% glucose in medium inhibited phenol degradation by ICBB 1170. Cell-free extracts of ICBB 1170 had specific activity 2.92 Ulmg. Degradation of benzoate by ICBB 1170 wasstudied. However, the ability to degrade phenol were higher than that of benzoate.
Chemoreaction Drying of Saccharomyces cerevisiae Culture with CaO and the Influence of Moisture Sorption Upon Stress and Death of the Dried Culture ., Novelina; Soekarto, Soewarno T; Jenie, Betty Sri Laksmi; Saono, Susono; Suhartono, Maggy T
Jurnal Teknologi Dan Industri Pangan Vol 16, No 1 (2005): Jurnal Teknologi dan Industri Pangan
Publisher : Departemen Ilmu dan Teknologi Pangan, IPB Indonesia bekerjasama dengan PATPI

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (467.664 KB) | DOI: 10.6066/473

Abstract

The aim of the research was to study chemoreaction drying of Saccharomyces cerevisiae culture and to a analyze the influence of moisture sorption on pattern of viability, stress and mortality of the dried starter culture. The cell culture of S. cerevisiae was produced in glucose media with aerobic process in a fermentor at 30C. The cell obtained was dried in a using chemoreaction process. Drying was conducted by coating with various thickness of coating and various ratio of CaO and calcium oxide ratio used in drying. Dry culture of S. cerevisiae and protective agent of CMC and jelly were added to the dry culture. The mixtrure then was stored at in decicators at R H 11 up to 97% until water equilibrium was achieved. Later the patterns of stress from each sample of dry culture at various conditions of moisture was analyzed. The result of the research showed that the best drying method was using coat thickness of 1.3 mm and calcium oxide 10 times the weight of dried sample. The condition of dry culture at low minimal water from 5 % were mostly in dormant state, the highest viability was at the rate of moisture of 5 % - 8 %, but a lot portion of the cells were inactive, and at the moisture higher than 8%, dead cells were observed. Addition 2% jelly of 2% CMC did not protect cells from stress. Key words: Saccharomyces cerevisiae, chemoreaction drying, moisture sorption, dry stresses
Anti Cancer Activity of Chitooligomers Wahyuni, Sri; Zakaria, Fransisca; Witarto, Arief B; Syah, Dahrul; Suhartono, Maggy T
Jurnal Teknologi Dan Industri Pangan Vol 17, No 1 (2006): Jurnal Teknologi dan Industri Pangan
Publisher : Departemen Ilmu dan Teknologi Pangan, IPB Indonesia bekerjasama dengan PATPI

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (532.143 KB) | DOI: 10.6066/406

Abstract

The chitin obtained from the crab industries can be used as a source for production of chitooligomers which has an important biological activity. The aims of this research was to evaluate anti cancer activity of the chitooligomers obtained from enzymatic hydrolysis using chitosanase from thermophilic bacterium Bacillus licheniformis MB2 isolated from Tompaso Manado. Media for producing the enzyme contained colloidal chitosan 1% and the enzyme was harvested after seven days of incubation at 550C. The heat stable protein enzyme was coagulated with 80% saturated ammonium sulphate and purificated using hydrophobic interaction chromatography with butyl sepharose gel. Enzyme of 0.005, 0.0085, 0.10 dan 0,17 IU/mg chitosan on soluble chitosan 1% substrate with 85% degree of deacylation were used to produce chitooligomers through incubation for one and three hours. The reaction products were analyzed (and fractionated) using HPLC. The effect of this samples on cancer cells was evaluated using K562 cells (chronic myelogenous leukemia) and investigated after being treated with MTT (3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide). In general, hydrolysates and fractionated chitooligomers showed better anti cancer activity than the 2- Bromo deoxy uridine used as positive control at similiar concentration (17 ?g/ml). Both of hydrolysates and fractionated chitooligomers (trimer to hexamer) inhibited proliferation of human K562 cancer cells line in vitro about 20.57% and 15.68% respectively.The apoptosis phenomena was found on K562 cells treated with chitooligomer hydrolysate which can be examined by Hoechts staining fluorescent method. Chitooligomers hydrolysate showed anti metastatic potential, the chitooligomers were found also as potent protease inhibitor. Keywords : chitooligomers, chitosan, anticancer
Cloning, Sequencing and Characterization of The Xylan Degrading Enzymes from Geobacillus thermoleovorans IT-08 Puspaningsih, Ni Nyoman Tri; Suwanto, Antonius; Suhartono, Maggy T
Jurnal ILMU DASAR Vol 9, No 2 (2008)
Publisher : Fakultas MIPA Universitas Jember

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (305.475 KB)

Abstract

Geobacillus thermoleovorans IT-08 is a Gram positive, thermophilic bacterium that can utilize xylan as a sole source of carbon. This strain was isolated from Gunung Pancar hot spring, Bogor, West Java, Indonesia. A plasmid genomic library in Escherichia coli DH5α was constructed and screened for xylanase activity. One positive clone, namely DH5α (pTP510) has been isolated, sequenced and showed putative exo-xylanase (exo-xyl), β-xylosidase (xyl), and α-L-arabinofuranosidase (abfa) genes (Genebank Accession No.DQ387047, DQ345777 and DQ387046 respectively). Each gene encoded 604, 511 and 502 amino acids, respectively. The BLAST search for protein database revealed that Abfa was high similar with GH51 family Abfa of Geobacillus stearothermophilus T6, but Xyl and Exo-Xyl were slight similar with GH43 family (25-34%) respectively. The deduced protein had a molecular weight of about 70 kDa (Exo-Xyl), and 60 kDa (Xyl and Abfa). These showed good accordance with the calculated molecular weight of each protein (68.64 kDa for Exo-xyl, 57.99 kDa for Xyl and 57.03 kDa for Abfa) from deduced amino acid sequence.Keywords : Thermophile xylanases, Geobacillus thermoleovorans IT-08, pTP510
Karakteristik Endapan Cairan Rumen Sapi asal Rumah Potong Hewan sebagai Feed Supplement Budiansyah, Agus; Resmi, Resmi; Nahrowi, Nahrowi; Wiryawan, Komang G; Suhartono, Maggy T; Widyastuti, Yantyati
Jurnal Ilmu-Ilmu Peternakan JIIP Volume XIV No. 1 Mei 2011
Publisher : Jurnal Ilmu-Ilmu Peternakan

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (71.575 KB)

Abstract

The aims of this experiment were to  identify and characterize of sediment product from cattle rumen liquor as a source of amino acids, minerals and vitamins. The sediment were obtained as pellets upon centrifugation of rumen liquor at 10,000 g for 10 minutes for collection of supernatant. The sediment were evaluated for amino acids, minerals and vitamins composition and chararacterized for the pH,  solubility of dry matter, specific density, bulk and compacted bulk densities and angle of response. Result of the experiment showed that sediment contained higher minerals:  Na, K and Fe compared with the commercial premix, but lower in B-vitamins and amino acids. The composition of mineral Na, K, and Fe from rumen liquor of local cattle was 13.47%, 7.73 % and 14.52 %,  while Na, K, and Fe from  rumen liquor of  imported cattle was 18.40%, 10.25%, and 14.52% respectively. The sediment had pH range from 10.01-10.03, the dry matter solubility  was 35.5% up to 39.1%. The sediment from imported cattle had higher specific density, bulk and compacted bulk densities and angle of response than that of local cattle. It is concluded that sediment from cattle rumen liquor contained high Na, K and Fe, low amino acids and B-vitamin, high pH and low solubility.
Isolasi dan kloning fragmen cDNA dari tanaman karet dengan sifat resisten dan rentan terhadap Corynespora cassiicola menggunakan metode cDNA-AFLP Isolation and cloning of cDNA fragments from rubber plant with resistant and susceptible characters to Corynespora cassiicola using cDNA-AFLP method NURHAIMI-HARIS, .; SUWANTO, Antonius; SUHARTONO, Maggy T; ASWIDINNOOR, Hajrial
E-Journal Menara Perkebunan Vol 78, No 1: Juni 2010
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (486.561 KB) | DOI: 10.22302/iribb.jur.mp.v78i1.78

Abstract

AbstractLeaf fall disease caused by Corynespora cassiicola fungi is one of the most important diseases in rubber plant. Rubber clone AVROS 2037 is considered resistant to this pathogen while clone PPN 2444 is susceptible. These two rubberclones were used to identify genes or transcripts differentially expressed during interaction between rubber plants and the fungi, using cDNA-Amplified Fragment Length Polymorphism (cDNA-AFLP) method. Induced genes/transcripts expression was examined to compare differencies between plants uninfected and infected with C. cassiicola at 24, 36, 48 and 72 hours after inoculation. cDNA-AFLP analysis was performed using restriction enzyme VspI and TaqI so the adapters and primers also have the recognition site of these enzymes. By using 29 specific primers, 35 out of approximately 1450 fragments were differentially expressed between AVROS and PPN 2444 clones. All of these fragments were cloned and sequenced. The homology-based grouping of these sequences resulted in 19 contigs and nine individual sequences. Among these, 10 contigs and five sequences have significant sequence homology with known genes in gene bank data base, such as Ran binding protein, protein transporter and transcriptional regulators of some organisms; arginase, GTP-binding protein, heat shock protein (HSP) and aconitase. Ran binding protein, GTPbinding protein and protein transporter were known as membrane proteins while arginase and HSP usually expressed as a response to wounding or toxin treatment. The present of arginase is usually related to the availability of nitric oxide (NO) in plant tissue. NO is well known as a signal molecule on plant defense response. AbstrakPenyakit gugur daun yang disebabkan oleh fungi Corynespora cassiicola merupakan salah satu penyakit penting tanaman karet. Klon karet AVROS 2037menunjukkan sifat resisten terhadap patogen tersebut sedangkan klon PPN 2444 merupakan klon yang rentan. Kedua klon karet tersebut digunakan untuk mengidentifikasi gen atau transkrip yang diekspresikan secara diferensial selama terjadi interaksi antara tanaman karet dengan C. cassiicola menggunakan teknik cDNA-Amplified Fragment Length Polymorphism (cDNA-AFLP). Ekspresi gen/transkrip dipelajari untuk membandingkan perbedaan antara tanaman yang tidak diinfeksi dengan yang diinfeksi patogen pada waktu 24, 36, 48 dan 72 jam setelah inokulasi. Analisis cDNA-AFLP dilaksanakan dengan menggunakan pasangan enzim restriksi VspI dan TaqI sehingga adapter dan primer memiliki sekuen pengenalan kedua enzim restriksi tersebut. Dengan menggunakan 29 pasang primer spesifik, sebanyak 35 dari sekitar 1450 fragmen memiliki ekspresi berbeda antara klon AVROS 2037 dan PPN 2444. Semua fragmen yang berbeda tersebut kemudian diklon pada vektor kloning dan disekuen. Hasil sekuensing dikelompokkan berdasarkan homologi sekuennya dan menghasilkan 19 contigs serta sembilan macam sekuen yang tidak mengelompok. Sebanyak 10 dari 19 contigs dan lima dari sembilan sekuen tersebut memiliki homologi dengan produk gen yang telah dikenal yang terdapat di pangkalan data GenBank, seperti putative Ran binding protein, protein transporter, regulator transkripsi, arginase, GTP-binding protein, heat shock protein (HSP) dan aconitase. Beberapa di antaranya seperti putative Ran binding protein, protein transporter dan GTP-binding protein dikenal sebagai protein membran, sedangkan arginase dan HSP merupakan protein atau enzim yang ekspresinya pada tanaman antara lain dipengaruhi oleh adanya pelukaan dan perlakuan toksin. Keberadaan arginase sering berhubungan dengan ketersediaan nitric oxide (NO) pada jaringan tanaman. NO dikenal sebagai salah satu sinyal molekul dalam mekanisme pertahanan tanaman.
Molecular analysis of Hemagglutinin Gen of Highly Pathogenic Avian Influenza of H5N1 Subtype Isolated from Waterfowls Susanti, R; Soejoedono, Retno D; K Mahardika, I Gusti Ngurah; T Wibawan, I Wayan; Suhartono, Maggy T
Indonesian Journal of Animal and Veterinary Sciences Vol 13, No 3 (2008)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14334/jitv.v13i3.584

Abstract

Avian influenza viruses (AIV) subtype H5N1 isolated from waterfowls in West Java pose the known characteristic of highly pathogenic strains, with polybasic amino acid sequence of cleavage site QRERRRKKR and QRESRRKKR. This research aimed to analyze the important domains of hemagglutinin (HA) gene of those isolates. Fragment of HA gene was amplified using RT-PCR method with specifically-designed primer pairs and sequenced using dideoxy termination method with ABI automatic sequencer (Applied Biosystems). Multiple alignment of nucleotide and deduced amino acid sequences were analyzed using ClustalW of MEGA-3.1 program. Some of biological domains of HA, i.e. antigenic sites, receptor binding pocket, and glycosylation sites of the isolates were polymorphic. The viruses also pose conserved glutamine (Q) and glisin (G) residues at the known receptor binding site, at the position 222 and 224 respectively. This findings clearly show that all AIV subtype H5N1 isolaed from waterfowl preservesthe α-2,3NeuAcGal avian receptor specificity. Key Words: Antigenic Sites, Glycosylation Sites, Receptor Binding Pocket, AIV H5N1, Waterfowls
Cloning, Sequencing and Characterization of The Xylan Degrading Enzymes from Geobacillus thermoleovorans IT-08 Puspaningsih, Ni Nyoman Tri; Suwanto, Antonius; Suhartono, Maggy T
Jurnal ILMU DASAR Vol 9 No 2 (2008)
Publisher : Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Jember

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (305.475 KB)

Abstract

Geobacillus thermoleovorans IT-08 is a Gram positive, thermophilic bacterium that can utilize xylan as a sole source of carbon. This strain was isolated from Gunung Pancar hot spring, Bogor, West Java, Indonesia. A plasmid genomic library in Escherichia coli DH5α was constructed and screened for xylanase activity. One positive clone, namely DH5α (pTP510) has been isolated, sequenced and showed putative exo-xylanase (exo-xyl), β-xylosidase (xyl), and α-L-arabinofuranosidase (abfa) genes (Genebank Accession No.DQ387047, DQ345777 and DQ387046 respectively). Each gene encoded 604, 511 and 502 amino acids, respectively. The BLAST search for protein database revealed that Abfa was high similar with GH51 family Abfa of Geobacillus stearothermophilus T6, but Xyl and Exo-Xyl were slight similar with GH43 family (25-34%) respectively. The deduced protein had a molecular weight of about 70 kDa (Exo-Xyl), and 60 kDa (Xyl and Abfa). These showed good accordance with the calculated molecular weight of each protein (68.64 kDa for Exo-xyl, 57.99 kDa for Xyl and 57.03 kDa for Abfa) from deduced amino acid sequence.