Articles

Bioenergetic Consequences of FLAG Tag Addition to the C-Terminus of Subunit 8 of Yeast Saccharomyces cerevisiae Mitochondrial ATP Synthase ARTIKA, I MADE
HAYATI Journal of Biosciences Vol 17, No 3 (2010): September 2010
Publisher : Bogor Agricultural University, Indonesia

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Abstract

The yeast mitochondrial F1F0-ATP synthase is a multisubunit complex that contains at least 17 different subunits. Subunit 8 of yeast mitochondrial ATP synthase is a hydrophobic protein of 48 amino acids encoded by the mitochondrial ATP8 gene. Subunit 8 has three distinct domains; an N-terminal domain, a central hydrophobic domain and a C-terminal domain. FLAG tag addition to subunit 8 protein potentially facilitate elucidation of its topology, structure, and function. It has been shown that following incorporation of FLAG tag to its C-terminus, subunit 8 still assemble into functional ATP synthase complex. In order to analyze bioenergetic consequences of the FLAG tag addition, a yeast strain expressing FLAG tagged-subunit 8 was subjected to cellular respiration assays. Results obtained showed that addition of FLAG tag to the C-terminus of subunit 8 does not impair its proper functioning. The FLAG tag system, therefore, can be employed to study subunit 8’s detailed structure, topology, and function.
Characterization of Bioflocculant Producing-Bacteria Isolated from Tapioca Waste Water SURYANI, .; AMBARSARI, LAKSMI; ARTIKA, I MADE; SUSANTI, HARTUTIK EKA
HAYATI Journal of Biosciences Vol 18, No 4 (2011): December 2011
Publisher : Bogor Agricultural University, Indonesia

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Abstract

Bioflocculant producing-bacteria from tapioca waste water were characterized. Two bacterial isolates i.e. LT-5 and LT-6 isolates had high flocculation activity, with activity of 68.92 and 71.38% respectively. The flocculation activity of LT-5 isolate increased at pH 2.0-4.0 (acidic condition), however the activity of LT-6 increased at pH 6.0-8.0 (neutral). Addition of 0.05% of AlCl3 as cation was the most effective and had important role in flocculation activity. Based on the morphological properties, LT-5 isolate was identified as Chromobacterium violaceum and LT-6 isolate was identified as Citrobacter koseri.
Development of Dual Control Allotopic Expression System for Subunit 8 of Yeast Saccharomyces cerevisiae Mitochondrial ATP Synthase ARTIKA, I MADE
HAYATI Journal of Biosciences Vol 18, No 3 (2011): September 2011
Publisher : Bogor Agricultural University, Indonesia

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Abstract

The yeast mitochondrial F1F0-ATP synthase is a multisubunit complex that contains at least 17 different subunits. Subunit 8 of yeast mitochondrial ATP synthase is a hydrophobic protein of 48 amino acids encoded by the mitochondrial ATP8 gene. A dual control allotopic expression system for subunit 8 has been developed. The strategy involves maintenance of two different compatible yeast expression vectors each utilizing a different inducible promoter in the same host cells. The system thus enables cloning and allotopic expression of two different forms of subunit 8 gene. The goal of the developed strategy is to permit allotopic expression of functional wildtype subunit 8 gene under a conditional promoter system and subunit 8 variant gene under the control of a different promoter system. The system is potentially useful for accurate assessment of assembly behavior of functionally defective subunit 8 variants in vivo. The strategy relies on the ability to conditionally regulate the expression of the two genes. A set of functionally defective subunit 8 variants has been cloned under an inducible yeast promoter system and dual plasmid harboring strains for dual control allotopic expression of each variant have been constructed.
Pemanfaatan Serbuk Kayu untuk Produksi Etanol dengan Perlakuan Pendahuluan Delignifikasi Menggunakan Jamur Phanerochaete Chrysosporium Irawati, Denny; Azwar, Norman Razief; Syafii, Wasrin; Artika, I Made
Jurnal Ilmu Kehutanan Vol 3, No 1 (2009)
Publisher : Jurnal Ilmu Kehutanan

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Abstract

Currently, Indonesia is in the middle ofpetroleum crisis. One ofthe alternative fuels which can be used as a petroleum substitute is ethanol. Ethanol can be produced from timber waste (sawdust). Indonesia in 2003 had timber waste potency of about 3-4 millions m3. However, ethanol production from sawdust has problems due to its lignin content. Therefore, research on bio-delignification treatment of sawdust prior to ethanol making process is required. In the present study ethanol was produced by simultaneous saccharification and fermentation (SSF) using crude cellulose from Trichoderma viride and Saccharomyces cerevisiae. The raw materials for ethanol production are sengon (Paraserianthes falcataria (L.) Nielsen syn.), meranti (Shorea sp.) and teak (Tectona grandis LIIVN.f.) sawdust after pretreatment with white rot fungi Phanerochaete chrysosporium for 10, 20 and 30 days incubation time. The yield of ethanol was between 1.65-44.83 g/1. The best combination treatment is sengon sawdust with 30 day incubation time.Keywords : sawdust, white rot fungi Phanerochaete chrysosporium, ethanol.
Antibacterial Activity of Propolis Produced by Trigona spp. Against Campylobacter spp. FATONI, AMIN; ARTIKA, I MADE; HASAN, AHMAD ENDANG ZAINAL; KUSWANDI, KUSWANDI
HAYATI Journal of Biosciences Vol 15, No 4 (2008): December 2008
Publisher : Bogor Agricultural University, Indonesia

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Abstract

Propolis is believed to have antimicrobial, anti-inflammatory and immunostimulating activities. The objective of this study was to investigate the antibacterial activity of ethanol extract propolis (EEP) of Trigona spp. from Bukittinggi West Sumatera against Campylobacter spp. Antibacterial activity of the EEP was measured by disc diffusion method. The compound groups of the propolis were also analyzed on the existence of alkaloids, flavonoids, saphonins, tannins, steroids, and terpenoids. This study revealed that the EEP of Trigona spp. shows an antibacterial activity on Campylobacter spp. The compound groups detected in the EEP were flavonoids and tannins, suggesting that the antibacterial activity of propolis of Trigona spp. may be due to these compounds. Key words: Trigona spp., antibacterial activity, Campylobacter spp
The Use of HIS6 Gene as a Selectable Marker for Yeast Vector ARTIKA, I MADE
HAYATI Journal of Biosciences Vol 16, No 1 (2009): March 2009
Publisher : Bogor Agricultural University, Indonesia

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Abstract

The yeast Saccharomyces cerevisiae HIS6 gene has been shown to be functional as a selectable marker for selecting and maintaining a yeast vector in yeast S. cerevisiae host cells. The yeast HIS6 gene encodes an enzyme involved in the yeast histidine biosynthesis. The yeast HIS6 gene was cloned into a yeast expression vector. The resultant recombinant plasmid was introduced into yeast host cells defective in endogenous HIS6 gene. The functionality of the HIS6 gene as a selectable marker was tested by growing transformed cells on selective minimum medium lacking histidine supplementation. Key words: HIS6 gene, selectable marker, vector, transformation, yeast
Fractionation and Characterization of Tannin Acyl Hydrolase from Aspergillus niger SANI ANWAR, YUNITA ARIAN; ARTIKA, I MADE; DANURI, HASIM
HAYATI Journal of Biosciences Vol 16, No 3 (2009): September 2009
Publisher : Bogor Agricultural University, Indonesia

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Abstract

We previously produced tannin acyl hydrolase (tannase) from Aspergillus niger isolated from cacao pod. In the present study the enzyme was subjected to fractionation by ammonium sulphate followed by dialysis process. The saturation level of ammonium sulphate used was 30-80% where the best enzyme activity was obtained at the saturation level of 60%. Compared to that of crude enzyme, specific activity of tannase after dialysis was four folds. Characterization results showed that optimum activity was at 35-50 oC and pH 6. Tannase was activated by K+ and Na+ at concentration of 0.01 and 0.05 M respectively. Mg2+ was found activate tannase only at 0.01 M. Addition of metal ions like Zn2+, Cu2+, Ca2+, Mn2+ and Fe2+ inhibited the enzyme activity. Kinetics analysis of various substrates tested showed that the Km value of tannic acid and gallotannin was 0.401 and 6.611 mM respectively. Vmax value of tannic acid was 10.804 U/ml and of gallotannin was 12.406 U/ml. Based on Michaelis-Menten constant (Km), the tannase obtained in the present study was more active in hydrolysing depside bonds rather than ester bonds. Key words: tannase, fermentation, fractionation, characterization, Aspergillus niger
Allotopic Expression of a Gene Encoding FLAG Tagged-subunit 8 of Yeast Mitochondrial ATP Synthase ARTIKA, I MADE
HAYATI Journal of Biosciences Vol 13, No 1 (2006): March 2006
Publisher : Bogor Agricultural University, Indonesia

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Abstract

Subunit 8 of yeast mitochondrial ATP synthase is a polypeptide of 48 amino acids encoded by the mitochondrial ATP8 gene. A nuclear version of subunit 8 gene has been designed to encode FLAG tagged-subunit 8 fused with a mitochondrial signal peptide. The gene has been cloned into a yeast expression vector and then expressed in a yeast strain lacking endogenous subunit 8. Results showed that the gene was successfully expressed and the synthesized FLAG tagged-subunit 8 protein was imported into mitochondria. Following import, the FLAG tagged-subunit 8 protein assembled into functional mitochondrial ATP synthase complex. Furthermore, the subunit 8 protein could be detected using anti-FLAG tag monoclonal antibody. Key words: allotropic expression, ATP synthase, mitochondria, yeast
Aktivitas Kitinase dan Peroksidase dari Ekstrak Kasar Protein Asal Kalus dan Berbagai Jaringan Tanaman Trichosanthes cucumerina var. anguina Sukma, Dewi; Poerwanto, Roedhy; Sudarsono, ,; Khumaida, Nurul; Artika, I Made; Wiyono, Suryo
Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy) Vol 40, No 3 (2012): JURNAL AGONOMI INDONESIA
Publisher : Bogor Agricultural University

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Abstract

Chitinase and peroxydase are important bioactive proteins or are specific enzymes that are related to plant resistance to pathogens. The aims of the research were to analyze the chitinase and peroxidase activities of crude protein extract from calli, stem, leaves and roots of T. cucumerinavar. anguina. In the first experiment, chitinase and peroxydase activities were analyzed from in vitro calli, leaves and roots obtained from 2-month-old of field grown plants. The media for calli induction were Murashige and Skoog medium with addition of 1 µM NAA + 1 µM BA, 2 µM NAA + 2 µM BA, 3 µM NAA + 3 µM BA, or 4 µM NAA + 4 µM BA. In the second experiment, the chitinase and peroxydase activities from crude protein extract of roots, stems and leaves were analyzed. The extracts were from 3-week-old seedling (less than a month), 1-month and 2-month-old plants. The first and the second experiment results showed that crude protein extracts of plant roots from the field grown plants had the highest chitinase and peroxidase activities. Stem of field grown plants had the similar level of chitinase activities with the plant roots. Chitinase activities of in vitro calli were not significantly different from those of plant roots so that it could be used as an alternative for plant roots in studying chitinase from T. cucumerinavar. anguina. Chitinase activities in crude protein extracts of roots appeared constant whereas peroxidase tend to increase with plant age. Keywords: calli, enzyme activities, leaves, roots, stem
Aktivitas Kitinase dan Peroksidase dari Ekstrak Protein Daun, Akar, Kalus dan Tunas In Vitro Trichosanthes tricuspidata Lour. Sukma, Dewi; Poerwanto, Roedhy; Sudarsono, ,; Khumaida, Nurul; Wiyono, Suryo; Artika, I Made
Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy) Vol 36, No 1 (2008): Jurnal Agronomi Indonesia
Publisher : Bogor Agricultural University

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Abstract

A number of Trichosanthes species has been reported as a source of bioactive protein associated with defense mechanisms such as chitinase. Chitinase and peroxidase of crude protein extracted from leaves, roots, in vitro calli and shoots of T. tricuspidata had been analysed. Calli were induced on MS medium containing combinations of 1 µM NAA + 1 µM BA  (K1), 2 µM NAA + 2 µM BA (K2), 3 µM NAA + 3 µM BA (K3), or 4 µM NAA + 4 µM BA (K4). Shoots were cultured in MS with 1 mg/l of BA, while leaves and roots were harvested from six-month old plants grown on the field. Results of the experiment suggested that K1-K4 medium could be used to induce calli although weight of calli from all medium composition  was not significantly different (0.19-0.31 g/explant/4 weeks). Calli from K1 medium had the highest of total crude protein content (3.24 mg/ml). The highest of chitinase activity was found in in vitro shoots (6.51 mM pNP/hour/mg protein) and the highest peroxidase activity was in the plant roots (0.25 ∆ 420/minute/mg protein).   Key words:  in vitro calli, shoots, crude protein, chitinase and peroxidase activities
Co-Authors , Sudarsono - Tri-Panji . HASIM . SURYANI A. E. Zainal Hasan Abdul Choliq Abidin, Syaeful ADINDA VIRGINIA DWI SETYO AHMAD ENDANG ZAINAL HASAN Akhmad Endang Zainal Hasan Amanda, Nisa Widya AMIN FATONI Antonius Padua Ratu, Antonius Padua Apon Zaenal Mustopa Aprianti, Apipah Arendra, Arya Arian, Perkasa Azmi Azhari, Azmi Budi Haryanto BUGI RATNO BUDIARTO Dedi Jusadi Denny Irawati Desi Purwaningsih DEWI SUKMA DIMAS ANDRIANTO DIMAWARNITA, Firda DWI N. SUSILOWATI Erismar Amri Evi Nur Qolbaini Febrianti, Fina H. A. E. Zainal Hasan Hami Seno, Djarot Sasongko HANIF, Hyakansa Hartono, Lusiana Kresnawati HARTUTIK EKA SUSANTI HARYANTO SUSILO HASIM DANURI Herti Sugiarti, Herti Ifa Manzila Iman Rusmana Inawati Inawati, Inawati Khusniati, Tatik Kuswandi . Kuswandi Kuswandi Laita Nurjanah, Laita LAKSMI AMBARSARI MARIA BINTANG Meilisza, Nina Mirza Dikari Kusrini MUHAMMAD AGUS SUPRAYUDI Muhammad Dailami, Muhammad MUHAMMAD ZAIRIN JUNIOR Norman Razief Azwar Novik Nurhidayat NUNUK WIDHYASTUTI Nur Bambang Priyo Utomo Nurmala Sari Nurul Khumaida PANJI, TRI - PUJI LESTARI Puspasari, Erna Rahadian Pratama, Rahadian Rahmawati, Fri Rahmi, Hayatul ROEDHY POERWANTO Safari, Dodi Septiany C. Palilingan Sheryn Sunni Albani Soekarno Mismana Putra, Soekarno Mismana SOGANDI, SOGANDI SUHARYANTO SUHARYANTO Suhyana, Jajang Sulistiani Sulistiani, Sulistiani Suryani Suryani SURYO WIYONO Sutoro Sutoro Syamsul Falah Tetty Chaidamsari, Tetty Tri Panji Trini Suryani Kadir Vita Rosaline Fahri Waras Nurcholis Wasrin Syafii Wijiastuti, Wijiastuti Yadi Suryadi YUNITA ARIAN SANI ANWAR YUNITA ARIAN SANI ANWAR