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Purification and Characterization of Streptomyces sp. IK Chitinase Margino, Sebastian; Nugroho, Agustinus Joko; Asmara, Widya
Indonesian Journal of Biotechnology Vol 15, No 1 (2010)
Publisher : Universitas Gadjah Mada

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Abstract

Streptomyces sp. IK isolated from compost inoculants, could produce extra cellular chitinase in a medium containing 0.2% (w/v) colloidal chitin, fermented for 96 hours at 30oC. The enzyme was purified by a combination of ammonium sulphate precipitation and DEAE-Cellulose anion-exchange chromatography. On SDS-polyacrylamide gel electrophoresis analysis, the purified enzyme showed a mass of 71 kDa. Chitinase was optimally active at pH of 6.7 and at 37oC. Km value and Vmax of the protein for colloidal chitin were 2.92 mg/ml and 4.26 ìg/h, respectively.Key words : chitinase, Streptomyces, purification, characterization
Human Origin Lactobacillus casei Isolated from Indonesian Infants Demonstrating Potential Characteristics as Probiotics in vitro ., Widodo; Taufiq, Tiyas Tono; Aryati, Ety; Kurniawati, Asih; Asmara, Widya
Indonesian Journal of Biotechnology Vol 17, No 1 (2012)
Publisher : Universitas Gadjah Mada

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Abstract

The aim of this experiment was to isolate and identify Lactic Acid Bacteria (LAB) from infant faeces and subsequent evaluation of its potential probiotics. LAB was isolated from faeces of infants who consumed breast milk as the only source of diet on L-cysteine-supplemented MRS Agar, and incubated on 37oC for 48 hours. Colonies grew on this media were then identifi ed based on morphological, physiological and molecular approaches. Morphological and physiological identifi cations based on Gram staining, shape, motility, spore formation, catalase, CO2 and NH3 production, and the ability to grow on temperature at 10oC and 45oC. Molecular identifi cation based on the amplifi cation of 16S rRNA gene. The potential application of selected isolates for probiotics was evaluated based on the ability to grow on media with low pH and the addition of 0.5% bile salts, the ability to inhibit the growth of pathogenic Bacillus cereus and Eschericia coli, and in vitroadherence ability. On the basis of morphological, physiological and molecular analysis of 16S rRNA gene, it was concluded that the selected isolate 1AF was a strain of Lactobacillus casei. Evaluation of probiotic in vitro showed that 60.4% of cells were resistant to pH 2.0 for 90 minutes. Survival of isolate 1AF after growing at 0.5% bile salts was 70.8%. The selected isolate 1AF showed the ability to inhibit the growth of Eschericia coli and Bacillus cereus with inhibitory zone of 12.00±1,00 and 15.33±1.53 mm, respectively. In vitro study on the adherence value of isolate to solid plate was found at 46.5%. It is concluded that Lactobacillus casei isolate 1AF is a potential candidate as probiotics and subject to further in vivo evaluation.
Chemosystematic of Enterobacteriaceae Familia Obtained from Blood Cultures Based on Total Protein Profiles Darmawati, Sri; Sembiring, Langkah; Asmara, Widya; Artama, Wayan T.; Anwar, Syaiful
Indonesian Journal of Biotechnology Vol 18, No 1 (2013)
Publisher : Universitas Gadjah Mada

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Abstract

The purpose of this study was to determine the chemosystematic of 14 strains of bacteria in blood cultures from Semarang using 1 reference strain S. typhi NCTC 786, based on the total protein profi les with the similarity relationship analysis based on Simple Matching Coeffi cient (SSM) analysis and algorithm methodof unweighted pair group with averages (UPGMA) presented in a dendrogram. The results showed that thechemosystematic based on the total protein profi les using SDS-PAGE method can classify the member ofbacterial strains of each species. The Clusters respectively consist of 4 strains of S. typhi (similarity: 89.7%),2 strains of Ser. marcescens (similarity: 89.7%), two strains of E. coli, and one strain of Salmonella ssp, S. typhi NCTC 786 (similarity: 100%). Those three incorporated clusters had the similarity value of 75.3%. Those four strains of Ent. cloacae composed in one cluster (similarity: 100%) are incorporated in a cluster consisting of one strain of Kleb. pneumoniae (similarity: 92.9%). Both clusters were incorporated in a cluster consisting of S. typhi NCTC 786 (similarity: 67.9%).Key words: Enterobacteriaceae, chemosystematic, blood cultures, protein profile
The Development of Pathogenicity of Avian Influenza Virus Isolated from Indonesia Wibowo, Michael Haryadi; Srihanto, Agus Eko; Putri, Khrisdiana; Asmara, Widya; Tabbu, Charles Rangga
Indonesian Journal of Biotechnology Vol 18, No 2 (2013)
Publisher : Universitas Gadjah Mada

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Abstract

Highly pathogenic avian infl uenza outbreak in Indonesia has been reported in various poultry due toH5N1 subtype. The presence of multiple basic amino acids within the cleavage site of HA glycoprotein hasbeen identifi ed to be associated with the pathogenicity of avian infl uenza virus. The study was retrospectivestudy which was designed to characterize the cleavage site and fusion site region of haemagglutinin gene ofAIV isolated from various poultry in 2003 to 2013. Isolation, Identifi cation and propagation were carried outto collect viral stock. For virus detection, reverse transcriptase PCR (RT-PCR) method on H5 and N1 genefragment was performed. All of RT-PCR HA gene positive products were sequenced for further nucleotideanalysis and to determine the nucleotide composition at the targeted fragment. The results are all AIV isolateswere identifi ed as H5N1 subtype. The sequence analyses revealed some motives of basic amino acid motivethat were classifi ed as highly pathogenic avian infl uenza virus. Further analyses on fusion domain of all AIVisolated during the period 2003 to 2013 showed conserved amino acid.Keywords: avian infl uenza, haemagglutinin, cleavage site, basic amino acid, fusion site
An Actinomycetes Producing Anticandida Isolated from Cajuput Rhizosphere: Partial Identification of Isolates and Amplification of pks-I genes ., Alimuddin; Asmara, Widya; Widada, Jaka; ., Mustofa; Nurjasmi, Reni
Indonesian Journal of Biotechnology Vol 15, No 1 (2010)
Publisher : Universitas Gadjah Mada

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Abstract

Actinomycetes have been the most prolific producer of various kinds of antifungal metabolites, and many of them are described as being produced by polyketide synthetases (pks). We present strain of Actinomycetes producing anticandida isolated from rhizosphere plant for amplification of Pks-I genes. The isolate was obtained from Wanagama I Forest UGM Yogyakarta. Gene of seven isolates, from total of 173 isolates, were amplified using degenerate primer to detect the presence of pks genes. One strain that is named Streptomyces sp. GMR-22 was partialy identified as anticandida producing actinomycete. The strain shown the strongest activity against Candida albicans. Based on bioautography assay, one spot active with Rf 0.57 was appeared as bright yellow by cerrium sulphate but it was and not visible on UV254 and 366 lights. Key words : pks genes, anticandida, Streptomyces sp GMR-22, rep-PCR, cajuput rhizosphere
Effect of Copper (II) Chloride Concentration towards Copper Ions Sorption on Natural Zeolite from Wonosari Irnawati, Dyah; Widjijono, Widjijono; Wijaya, Karna; Asmara, Widya
The Indonesian Journal of Dental Research Proceeding Book
Publisher : The Indonesian Journal of Dental Research

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Abstract

Copper-containing zeolite has been developed as an antibacterial material. It is obtained by reacting zeolite and salt solution with varied concentration of copper. Natural zeolite is abundant in Wonosari, Yogyakarta. The aim of this research was to investigate the concentration effect of copper (II) chloride solutions towards copper ions sorption on natural zeolite from Wonosari. Copper (II) chloride dihydrate powders, CuCl2. 2H2O, (Merck, Germany) and natural zeolite (Wonosari, Indonesia) were used. Zeolite powder (100 mesh) was washed and dried (200oC/2 hours). Copper (II) chloride solution with different concentrations (0.05 M, 0.10 M, 0.15 M, 0.20 M, and 0.25 M) were prepared. Zeolite and CuCl2 solutions (50 g/250 ml) were reacted at 60oC for 1 hour (n=5). The solutions were filtered, washed, and dried (100oC/24 hours). Copper ions amount were measured by XRF (Canberra Inc., USA). The data were analyzed by one way Anova. Mean values (% weight) of copper ions amount were 0.441 ± 0.029 % (0.05 M group), 0.588 ± 0.027 % (0.10 M group), 0.657 ± 0.014 % (0.15 M group), 0.676 ± 0.037 % (0.20 M group), and 0.737 ± 0.039 % (0.25 M group). The Anova showed that CuCl2 concentration influenced copper ions amount significantly (p<0.01). The differences among the groups were significant, except between 0.15 M and 0.20 M groups (p>0.05). Copper (II) chloride concentration influenced the copper ions sorption on natural zeolite from Wonosari. The 0.25M CuCl2 solution gave the highest copper ions amount.
The Phytochemical Analysis of Hay Infusions and Papaya Leaf Juice as an Attractant Containing Insecticide for Aedes Aegypti Cahyati, Widya Hary; Asmara, Widya; Umniyati, S Rahmah; Mulyaningsih, Budi
Jurnal Kesehatan Masyarakat Vol 12, No 2 (2017): Jurnal KEMAS Vol.12 No.2 : January 2017
Publisher : Jurusan Ilmu Kesehatan Masyarakat Fakultas Ilmu Keolahragaan

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15294/kemas.v12i2.6223

Abstract

Aedes aegypti mosquito population could be controlled by using lethal ovitrap. The addition of hay infusions as a attractant greatly enhance Aedes aegypti eggs trapped, and papaya leaf juice may inhibit Aedes aegypti eggs evolve to larvae or a larvae to pupae stage. This study was conducted to find out the chemical compounds in hay infusion and papaya leaf juice. We used phytochemical test using UV-Vis Spectrophotometry, Thin Layer Chromatography, and High Performance Liquid Chromatoraphy (HPLC) method. The results showed that hay infusion contains 12,75 mg/L ammonium and <1,20 ppm (µg/mL) lactic acid and papaya leaf juice contains 0,25% alkaloid, 0,14% flavonoid, 0,30% saponin, ≤68 mg/L steroid and 11,34% tannin, but negative terpenoid. We concluded that hay infusion and papaya leaf juice contains chemical compounds that could be use as attractant and bioinsecticide to Aedes aegypti, respectively.
Efek Jumlah Kandungan Filler Nanosisal Terhadap Ketahanan Fraktur Resin Komposit Nugroho, Dwi Aji; ., Wdjijono; ., Nuryono; Asmara, Widya; Wajar, Dony
Insisiva Dental Journal Vol 6, No 1 (2017)
Publisher : Insisiva Dental Journal

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Abstract

Latar Belakang: Salah satu komposisi resin komposit yaitu filler anorganik. Filler umumnya berasal dari glass.Material anorganik tersebut memiliki kekurangan yaitu proses pengolahan bersifat abrasive, polutan, dan konsumsienergi tinggi. Oleh karena itu, filler alternatif diperlukan, khususnya dari bahan organik. Bahan organik yangberpotensi adalah serat alam berupa sisal. Penelitian ini membuat bahan tumpatan resin komposit dengan sisalberukuran nano sebagai fillernya dan kita sebut nanosisal komposit. Tujuan Penelitian: untuk mengetahui pengaruhjumlah filler nanosisal terhadap ketahanan fraktur resin komposit. Metode Penelitian: Penelitian ini menggunakan 20sampel. Sampel yang digunakan adalah cetakan berbentuk balok berukuran 2x2x25mm (ISO-4049). Sampel dibagimenjadi empat kelompok dengan masing-masing berjumlah lima sample. Presentase filler masing kelompok adalah60%,65%,70% dan 3M ESPE-Z350. Sample diuji ketahanan fraktur menggunakan universal testing machine. Analisisdata menggunakan uji non-parametric Kruskal-Wallis. Hasil Penelitian: Resin komposit dengan filler 60% memilikirata-rata ketahanan fraktur 28.61 MPa, filler 65% sebesar 11,77 MPa, filler 70% 11,56 MPa dan resin komposit3M ESPE-Z350 35,36 MPa. Kruskal-Wallis menunjukkan tidak ada perbedaan signifikan (p>0,05). Kesimpulan:Berdasarkan hasil penelitian ini, tidak terdapat efek jumlah filler terhadap ketahanan fraktur resin komposit nanosisal.Kelompok resin komposit 3M ESPE-Z350 memiliki nilai ketahanan fraktur tertinggi. Jumlah kandungan filler yangoptimal dalam komposit nanosisal adalah 60%.
Karakterisasi protease Bacillus sp. UGM5 Widowati, Titik Purwati; Djojowidagdo, Soemitro; Setiyono, Setiyono; Asmara, Widya
Majalah Kulit, Karet, dan Plastik Vol 14, No 26 (1999): Majalah Barang Kulit, Karet, dan Plastik
Publisher : Center for Leather, Rubber, and Plastic Ministry of Industry, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1359.555 KB) | DOI: 10.20543/mkkp.v14i26.297

Abstract

The objective of this experiment is to indentify the characters of proetease produced by Bacillus sp.UGM5.the protease secreted by Bacillus sp.UGM5 was first isolated,purified and then charactirezed.The crude enzyme has spesific actifity of 1.14 U/mg,however,the spesific activity of purified enzyme was increased by 23.8 times fold and recovery was 33.69%.The Page of nondenatured crude enzymes showes two type of proreases,however ,the SDS-Page of denatured purified enzyme showed four protein-bends with molecular weights of 55.5 kDa,18kDa respecetively.The optimum pH and temperature for the enzyme acrivity are 8.5 and 420C and belongs to serin protease type,with Km 3 X 10-3mM and Vmax 0.0890mM/30 minutes.The activity is not inhibited by Ca+2,Fe+2 and EDTA. INTISARITujuan penelitian ini adalah untuk mengkarakterisasi  protease yang diproduksi  Bacillus sp. UGM5. Protease yang disekresi oleh Bacillus sp.UGM5 pertama-tama diisolasi,dipurifikasi kemudian dikarakterisasi.Enzim kasar mempunyai spesifik 1,14 U/mg.Aktivitas spesifik enzim yang dimurnikan meningkat 23,8 kali dengan recovery 33,69%.Elektroforesis gel poliakrilamid enzim yang dipurifikasi dan didenaturisasi menunjukan ada empat pita protein dengan berat protein masing-masing 55,5 kDa,51 kDa,18 kDa,dan 15,5 kDa .ph dan temperatur optimum aktivitas enzim adalah 8,5 dan 420C,termasuk tipe protease serin,dengan nilai Km 3 X10-3mM dan Vmax 0,0890mM/30menit.Aktivitas enzim tidak dihambat oleh adanya ion Ca2+,Fe+3 dan EDTA.
PURIFICATION AND CHARACTERIZATION OF THE NEWLY THERMOSTABLE PROTEASE PRODUCED BY Brevibacillus thermoruber LII ISOLATED FROM PADANG CERMIN HOTSPRING, INDONESIA Zilda, Dewi Zeswita; Harmayani, Eni; Widada, Jaka; Asmara, Widya; Irianto, Hari Eko; Patantis, Gintung; Fawzya, Yusro Nuri
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 9, No 1 (2014): May 2014
Publisher : Research and Development Center for Marine and Fisheries Product Processing and Biotechnol

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/squalen.v9i1.91

Abstract

Thermo stability is among of the vital enzyme characteristics for industrial application. Brevibacillus thermoruber LII was obtained as a potential isolate from the previous researchwhich screened the potential thermostable protease producing bacteria from Indonesian hotspring.The newly thermostable protease produced by thermophilic Brevibacillus thermoruber LII hadbeen purified and characterized. It was predicted that the pure enzyme obtained from Brevibacillusthermoruber LII was homo hexameric, having molecular weight of 36 kDa unit protein and itsnative was 215 kDa. In addition, it was also a neutral metalo serine protease according tobiochemical tests that it was totaly inhibited by PMSF (Phenylmethanesulfonyl fluoride) and EDTA(Ethylenediaminetetraacetic acid). It showed optimum activity at pH of 8 and active in acidic buffer(up to pH of 4). All of metal ion in the form of chloride salt (2.5 mM) which were tested on theenzyme enhanced the enzyme activity but Li2+. Ca2+ion increased the activity and the stability ofenzyme against thermal. The enzyme also showed the stability against solvent. The protease LIIhad optimum temperature at 60oC without CaCl 2and 80 – 85oC with addition of 2.5 mM CaCl 2. TheK Mand V maxvalues for the purified protease LII were 27.2 mg/ml or 0.362 – 0.272 M for substrateHammersteinCasein (MM 75–100 kDa) and 261.1 µg/minute/ml, respectively.
Co-Authors . Sismindari ., Wdjijono A. Alimuddin Adi Heru Sutomo Aditya Krishar Karim Aditya Krishar Karim AETH. Wahyuni AETH. Wahyuni, AETH. Ag. Yuswanto Agnes Sri Harti Agnes Sri Harti Agnesia Endamg Tri Hastuti Wahyuni Agnesia Endang Tri Hastuti Wahyuni Agus Eko Srihanto Agus Eko Srihanto, Agus Eko Agustinus Joko Nugroho Agustinus Joko Nugroho, Agustinus Joko Aisida, Sufinatin Alma Linggar Jonarta Ambar Pertiwiningrum Andini, Syarifah Anwar Rosyidi April H Wardhana Arfian Rahma Shanti Artama , Wayan T. Arum Setiawan Arum Setiawan Asih Kurniawati Bambang Hariono Bambang Sumiarto Bambang Sutrisno Bancin, Girang Stevani Banun Kusumawardani Boy Bachtiar Budi Mulyaningsih Budi Mulyono BUDI SETIADI DARYONO Budi Setiadi Dayono Charles Rangga Tabbu Charles Rangga Tabbu Chatarina Behar Chatarina Behar, Chatarina Darmawati , Sri Darmayanti, Hervina Puspita Dewi Noor Hidayati Dewi Seswita Zilda Dito Anggoro Djaswadi Dasuki Doddi Yudhabuntara Dwi Aji Nugroho, Dwi Dyah Haryuningtyas Dyah Haryuningtyas Dyah Irnawati Eko Agus Srihanto Eko Agus Srihanto Eko Agus Srihanto, Eko Agus Elfayetti Elfayetti Elsa Kardiana Eni Harmayani Eni Harmayani Enny Yusuf Wachidah Yuniwarti Ety Aryati Ety Aryati, Ety Febriani Putri, Devita Fiska Salsabilla Gaol, Riski Fanni Lumban Gintung Patantis Gintung Patantis Gusti Ayu Yuniati Kencana Hafiza, Rohmania Zuhro Hamzah Muhammad Mardi Putra hanifah hanifah Hanny Hafiar Hardyanto Soebono Hardyanto Subono Harefa, Meilinda Suriani Harefa, Sri Aswinda Hari Eko Irianto Hasibuan, Novita Hasibuan, Novita Annisah Heru Susetya Hutauruk, Mutiara Cristeofani I Gusti Ketut Suarjana I Gusti Made Krisna Erawan I Gusti Ngurah Kade Mahardika I Wayan Suardana I. Istriyati Ida Tjahajati Iisnawati, Iisnawati Ika Dyah Kusumawati Ika Dyah Kusumawati, Ika Dyah Indwiani Astuti Istriyati ., Istriyati Istriyati Istriyati Istriyati Istriyati Istriyati, . Istriyati, . Iwan Dwiprahasto JAKA WIDADA Juni Handajani Kardians, Elsa Karna Wijaya Kerin Sisca Octaviani Luahambowo Khairunnisa Khairunnisa Khairunnisa Nasution Khrisdiana Putri Khrisdiana Putri, Khrisdiana Koichi Akiyama Krisdiana Putri Kumalasari, Eliana Dwi Langkah Sembiring LANGKAH SEMBIRING Langkah Sembiring Lina Sri, Widyawati Liza Angeliya M. Haryadi Wibowo M. Mustofa Mammed Sagi Mammed Sagi Mangkoewidjojo , Soesanto Marla Anggita Marpaung, Viviana Marsetyawan Soesatyo Marsetyawan Soesatyo Masashi Kawaichi Masashi Kawaichi, Masashi Maxs Urias Ebenheizer Sanam Meilinda Suriani Harefa Michael Haryadi Wibowo MM.Firdiana Krisnaningsih Mustofa M, Mustofa Mustofa Mustofa Niken Irfa Nastiti Ning Rintiswati Ning Rintiswati Nobuyuki Harada Nursyirwani, Nursyirwani Nurul Islamiyah Nuryono ., Nuryono Nyoman Reishita Andriyani Osman Sianipar Prasetyo, Dimas Bayu Purba, Tondang Raja Pangihutan Purnama Edy Santosa Purwoko, Agus Putra, Mulhady Raden Wisnu Nurcahyo Rahma, Mawaddah Rahmat Setya Adji Rahmat Setya Adji Regina TC Tandelilin Reni Nurjasmi Reni Nurjasmi, Reni Rini Widayanti Risbue Siregar Rosa Delima Lumbantungkup S Muharsini S Rahmah Umniyati, S Rahmah S. Sismindari Sabela, Serli Sagi , Mammed Salomo Hutahaean Salsabilla, Friska Santoso, Ferdinand Prayogo Cahyo Sarwo Edy Wibowo SATRIYAS ILYAS Sebastian Margino Sebastian Margino Sembiring , Langkah Sembiring, Samariana Br Setiawan , Arum Setiyono Setiyono Setiyono Setiyono Setyawan Budiharta Sidna Artanto Sidna Artanto Sidna Artanto, Sidna Simaremare, Ermas Simarmata, Claudia Grace Siregar, Risbue Sismindari . Siti Sunarintyas Sitti, Ummiyati Rahmah Soemitro Djojowidagdo Soesanto Mangkoewidjojo Sri Darmawati Sri Darmawati Sri Darmawati Sri Lestari Subronto Prodjoharjono Sugeng, Mardihusodo Juwono Sugiharto Sulastri, Yati Surya Amanu Surya Amanu Surya Amanu Surya Amanu Susi Iravati Susi Iravati Susi Maulida, Susi Syaiful Anwar Syari’I Damanik, Muhammad Ridha Syukri Hidayat Titik Purwati Widowati Titik Purwati Widowati, Titik Purwati Tiyas Tono Taufiq Tondang Raja Pangihutan Purba Tri Ratnaningsih Tri Untari Tri Untari Tri Untari Tri Wibawa Triyanto . Tsutomu Nohno Vivina Marpaung W. Widjijono W. Widodo Wajar, Dony wayan T Artama wayan T Artama Wayan T. Artama Wayan T. Artama Wayan T. Artama Wayan Tunas Artama Wayan Tunas Artama Widagdo Sri Nugroho Widagdo Sri Nugroho Widodo Widodo Widya Hary Cahyati Wisnu Nurcahyo Wulandari, Anna Yahya, Adibah Yatri Drastini Yulita Kristanti Yundari, Yundari Yuni Wijayanti Yusro Nuri Fawzya Yusro Nuri Fawzya Zahwa, Afitzka Al Zilda, Dewi Zeswita