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Erda Eni Rame Hau
Politeknik Pertanian Negeri Kupang
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OPTIMAL PROTEIN Cj0391c EXPRESSION AND PURIFICATION METHODS FOR PROTEIN IDENTIFICATION Erda Eni Rame Hau
Partner Vol 24, No 1 (2019): Edisi Juli
Publisher : Politeknik Pertanian Negeri Kupang

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35726/jp.v24i1.331

Abstract

Identifikasi mekanisme virulensi yang terkait dengan Campylobacter jejuni adalah bidang yang penting karena merupakan target potensial untuk vaksin masa depan. Contoh protein yang disekresikan oleh C. jejuni yang telah dipelajari baru-baru ini adalah Cj039lc.  Analisa bioinformatika menemukan bahwa protein ini bisa menjadi ?-helical pore forming protein (PFP).  Untuk mengetahui lebih lanjut mekanisme virulensi dari protein ini maka perlu dilakukan identifikasi lebih mendalam tentang protein.  Oleh karena itu penelitian ini bertujuan untuk mengidentifikasi metode ekspresi dan purifikasi yang optimal untuk menghasilkan protein Cj3091c yang murni dan cukup, yaitu 1 mg/ml Cj0391c murni.  Pemurnian dilakukan dengan Iminobilized metal Affinity Chromatography (IMAC) kolom yang digunakan untuk memulihkan protein rekombinan. SDS-PAGE dan Western blotting digunakan sebagai metode untuk menentukan kondisi optimum untuk elusi protein rekombinan Cj039lc.  Hasil dari ekspresi dan pemurnian pada percobaan ini menyimpulkan bahwa konsentrasi optimum Imidazole dalam buffer pencuci untuk menghasilkan konsentrasi tinggi protein Cj039lc murni adalah kombinasi dari langkah awal 20 mM buffer pencuci Imidazole diikuti dengan dua kali mencuci menggunakan 40 mM Imidazole. Konsentrasi akhir l mg/mL Cj039l c kemudian siap untuk digunakan untuk Small-angle X-ray scattering (SAXS) dan Dynamic light scattering (DLS).
PROTEIN Cj0391c INTERACTION WITH LIPID BILAYER MEMBRANE IDENTIFICATION USING SMALL ANGLE X-RAY SCATTERING (SAXS), WAXS AND DYNAMIC LIGHT SCATTERING (DLS) Erda Eni Rame Hau
Partner Vol 24, No 2 (2019): Edisi November 2019
Publisher : Politeknik Pertanian Negeri Kupang

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35726/jp.v24i2.379

Abstract

Compylobacter jejuni merupakan salah satu bakteri utama penyebab gastroenteritis akut di seluruh dunia dengan tingkat morbiditas dan mortalitas yang tinggi, terutama di negara-negara berkembang. Oleh karena itu sangat penting untuk mulai mengidentifikasi faktor virulensi dari C. jejuni, salah satunya adalah dengan mengidentifikasi protein yang dikeluarkan dari patogen ini.  Cj039lc merupakan salah satu protein yang telah mulai dipelajari dan dapat terindentifikasi sebagai ?-Helical pore toning protein (PFP) dan menyebabkan apoptosis pada sel makrofag ayam.  Karena diduga bahwa mekanisme apoptosis mungkin mirip dengan mekanisme cytotoxicity dari ?-Helical yang lain maka dalam penelitian ini protein Cj039lc ini kemudian diekspresikan, dimurnikan dan dianalisis untuk melihat interaksinya dengan membran lipid bilayer. Tujuan penelitian utama penelitian ini adalah untuk menentukan apakah Cj0391c bisa menembus dan mengganggu membran sebagai ?-PFP atau tidak.  Potensial interaksi protein dan membran dievaluasi menggunakan Small Angle X-Ray Scattering (SAXS), Wide Angle X-Ray Scattering (WAXS), dan Dynamic Light Scattering (DLS).  Meskipun hasil tidak menunjukkan indikasi penyisipan protein ke dalam membran, namun penelitian ini menjadi rujukan metode ekspresi dan pemurnian yang optimal untuk Cj0391c dan gambaran bentuk struktur Cj0391c berdasarkan hasil analisis SAXS.Keywords: Campylobacter jejuni, Cj039lc, pore forming protein, apoptosis, ?-PFP, lipid bilayer membrane, SAXS, WAXS, DLS.
GAMBARAN HISTOPATOLOGI GINJAL TIKUS (Rattus rattus) PADA PEMBERIAN LAMTORO MERAH (Acacia villosa) ADAPTASI DAN TANPA ADAPTASI Erda Eni Rame Hau
Partner Vol 21, No 1 (2016): Edisi Juli
Publisher : Politeknik Pertanian Negeri Kupang

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35726/jp.v21i1.142

Abstract

ABSTRACTIncubation of red acacia (Acacia villosa) leaf in preadapted rumen fluid of goat: Its effects on histopathologycal changes of kidney of rat (Rattus-rattus). This study was conducted to examine the detoxification potential of rumen microbes upon secondary compounds of Acacia villosa.  Leaf powder of A. villosa incubated in rumen fluid of goat pre-adapted with A. villosa was fed on rats used as experimental animals. Eigthteen rats were divided into 3 groups, consisting of control, group of rats fed with A. villosa incubated in pre-adapted rumen fluid (adaptation group) and group of rats fed with A. villosa incubated in non adapted rumen fluid (non-adaptation group).That results of histopathologycal examination showed a significant higher gromerular and tubular changes in the kidneys of the non-adaptation group compared to those in the adaptation and control groups. The changes in glomerulus were edema characterized by deposition of protein in the mesangial and Bowman space, and enlargement of the Bowman space. The changes in tubules were consisting of hydropic degeneration, deposition of protein in the tubules lumen and necrosis.Key words: Acacia villosa, histopathologycal changes, adapted rumen fluid.
GAMBARAN TOTAL PLATE COUNT (TPC) PADA DAGING SAPI YANG DIAMBIL DI RUMAH POTONG HEWAN (RPH) KOTA KUPANG Jois Moriani Jacob; Erda Eni Rame Hau; Yanse Yane Rumlaklak
Partner Vol 23, No 1 (2018): Edisi Juli
Publisher : Politeknik Pertanian Negeri Kupang

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35726/jp.v23i1.291

Abstract

The Increasing of beef consumption in Kupang City, shows that there is an increase in people's need for animal protein. However, high meat consumption can also be a threat to health if contaminated by microorganisms.This study aims to calculate the amount of microbial contamination in beef samples taken from Kupang City Slaughterhouse. 30 longissimus dorsi muscle samples were taken from RPH and taken to the Kupang City Veterinary Engineering Unit to be examined using the Total PlateCaount Test (TPC), then the bacterial colonies were calculated using a colony counter.The results of the TPC analysis show that 63.33% of the samples have TPC values that are above the normal standard, while only 36.67% are at normal values.
PENGARUH KONDISI DAN LAMA PENYIMPANAN PADA SUHU RUANG DAN REFRIGERATOR TERHADAP ANGKA TOTAL PLATE COUNT (TPC) SAMPEL SEI BABI DARI 4 TOKO DI KOTA KUPANG Erda Eni Rame Hau; Eny Rohyati
Partner Vol 23, No 2 (2018): Edisi November
Publisher : Politeknik Pertanian Negeri Kupang

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35726/jp.v23i2.328

Abstract

This study aims to evaluate the effect of different storage conditions and duration to the growth and development of microorganisms and pathogens up to a certain interval time. Therefore, the research will look at how the growth of bacteria in Se’i pork will be seen through the Total Total Count (TPC) in the condition of room temperature and refrigerator up to 5 days of storage time intervals. By treating the room temperature and temperature of the refrigerator with a storage time of 0, 1, 2, 3, 4, and 5 days, the microorganism numbers will be obtained in these two different storage conditions for 5 days.  The results showed clear result of the growth rate and the number of bacteria between Se’I stored at therefrigerator temperature storage with Se’I at room temperature every day.  It is clear from the results that the growth of microorganisms greater in Se'i at room temperature compared to those refrigerated. Other results found that samples from one shop have already shown TPC number above the maximum standard for the total number of microorganisms allowed by Standar Nasional Indonesia (SNI) in smoked meat.
INTERACTIONS OF OLIVE OIL POLYPHENOLS WITH AB OLIGOMERS Jois Moriani Jacob; Erda Eni Rame Hau
Partner Vol 24, No 2 (2019): Edisi November 2019
Publisher : Politeknik Pertanian Negeri Kupang

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35726/jp.v24i2.387

Abstract

Neurotoksisitas terutama mempengaruhi patogenesa penyakit Alzheimer (AD). Mekanisme yang diprediksi adalah pembentukan dan pengendapan plak beta amiloid (AB) dalam keadaan agak terlarut dalam jaringan otak. Senyawa polifenol yang dihasilkan oleh minyak zaitun telah dihubungkan dengan treatment dari pembentukan plak AB dengan cara mengikat dan menghambat pembentukan agregat oligomerik beracun yang terlarut.  Studi ini bertujuan untuk menganalisis afinitas pengikatan AB (11-42)- oligomer dengan beberapa senyawa polifenol dan menentukan senyawa yang paling mampu untuk menghambat agregasi AB-oligomer. Metode komputasi digunakan untuk memprediksi kemampuan senyawa polifenol untuk berikatan dengan AB (11-42)-oligomer.  Metode ini termasuk docking, visualisasi 3D, dan analisis struktural.  Hasil menunjukkan hubungan positif yang kuat antara server docking dan skor afinitas pengikatan yang dihasilkan. Hasil lebih lanjut diilustrasikan polifenol dengan kelompok O-H tinggi dari struktur kimia mereka berkorelasi dengan ikatan afinitas yang lebih tinggi untuk AB (11-42)-oligomer.  Hesperidin memiliki jumlah kelompok O-H tertinggi dalam studi dan juga memiliki salah satu nilai ikatan afinitas tertinggi di ketiga server docking. Hal ini menunjukkan hubungan timbal balik antara struktur polifenol dan ikatan afinitas yang dihasilkan.
AKTIVITAS ANTIBAKTERI NIRA LONTAR TERFERMENTESI DENGAN VARIASI LAMA WAKTU FERMENTASI TERHADAP BAKTERI GRAM POSITIF (Staphylococcus aureus) DAN GRAM NEGATIF (Escherichia coli) Erda Eni Rame Hau; Eni Rohyati
JURNAL KAJIAN VETERINER Vol 5 No 2 (2017): Jurnal Kajian Veteriner
Publisher : FAKULTAS KEDOKTERAN HEWAN UNIVERSITAS NUSA CENDANA

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35508/jkv.v5i2.957

Abstract

This study aims to determine the effect of spontaneous fermented nira with different treatment time to gram positive and gram negative bacteria. The research was conducted in November 2017 at the Animal Health Laboratory, Kupang State Agricultural of Polytechnic and Veterinary Public Health Laboratory, Faculty of Veterinary Medicine, Nusa Cendana University, Kupang, NTT. Samples of bacteria used in this study were Staphylococcus aureus and Escherichia coli bacteria. The fermented of nira lontar from 4 fermentation time (1 day (N1), 2 days (N2), 3 days (N3), and 4 days (N4)) were then collected to test the antimicrobial activity in inhibiting the growth of the microorganism. Fermentation was carried out for 4 days because previous research showed that the optimum growth of lactic acid bacteria in fermented palm was on the 4th day. This study used the Kirby-Bauer disc method, in which antibacterial activity was described by the diameter of the inhibition zone. The results showed that the mean diameter of the inhibitory zone of S. aureus with N1 material was 8.15 mm, while N2, N3, and N4 had the same diameter of 6.4 mm. There was a significant difference between the diameter of the N1 zone and the rest of the samples, as well as all treatments with Ciprofloxacin positive control antibiotics. Different trends were indicated by E. coli, where there were no significant differences between N1, N2, and N3 diameters with values of 9.2 mm, 8.95 mm, and 8.1 mm, respectively, but they were significantly different from the diameter N4 of 6.4mm, and all treatments were significantly different with Ciprofloxacin positive control antibiotics. It can be concluded that spontaneous fermented nira lontar at four different fermentation time intervals have antimicrobial activity against gram-positive and gram-negative bacteria, with statistically significantly different inhibitory activity among different fermentation days. Moreover, fermented nira lontar has higher inhibitory activity on gram-negative bacteria, E. coli compared to gram-positive bacteria, S. aureus.
PENGARUH KEMASAN SERTA KONDISI DAN LAMA PENYIMPANAN TERHADAP PERTUMBUHAN Staphylococcus aureus PADA DAGING SEI BABI Erda Eni Rame Hau
JURNAL KAJIAN VETERINER PROSIDING SEMINAR NASIONAL KE-7
Publisher : FAKULTAS KEDOKTERAN HEWAN UNIVERSITAS NUSA CENDANA

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35508/jkv.v0i0.1596

Abstract

In addition to be sold freshly for consumption after being processed, se’í babi which is a popular meat product of Kupang City has also been given a vacuum packing processing with the aim to extend the shelf life of meat. This research aims to determine the effect of the packaging and storage conditions of seí babi from four meat shops in the Kupang to the growth and number of pathogenic microorganisms Staphylococcus aureus up to the some certain time. S. aureus is one of foodborne disease bacteria that can contaminate meats. Bacterial contamination can come from the environment or bacterial infections in farm. Total of 16 se’i babi samples were used in this research, 2 samples from each meat shops. Two different packaging samples from all shops were then stored in two different conditions (room temperature and refrigerator). Samples were then being proceeded for S. aureus microbial evaluation every day since day-0 to day-5. Samples were cultured on Baird Parker medium and Mannitol Salt Agar and followed by bacteria identification by Gram stain, catalase test, oxidase test, and coagulase test. The number of S. aureus colonies was calculated and the result was compared to the limit of microbial contamination based on Standar Nasional Indonesian (SNI). The number of S. aureus from day 1 until day 5 of all samples were > 1 × 102 CFU/g, more than SNI (7388:2009). The results showed differences in the total number of bacteria between meats on refrigerator temperature storage, which is in general is lower compared to meat in room temperature storage. The same trend also occurs in vacuum packed meats, they had lower number of S. aureus compared to samples without vacuum packed.
Co-Authors Eni Rohyati, Eni Eny Rohyati Jois Moriani Jacob Rumlaklak, Yanse Yane