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All Journal VALENSI Bionatura Teknotan: Jurnal Industri Teknologi Pertanian Jurnal Natur Indonesia Microbiology Indonesia Majalah Kedokteran Bandung Indonesian Journal of Pharmaceutical Science and Technology LEMMA: Letters Of Mathematics Education Indonesian Journal of Biotechnology QALAMUNA: Jurnal Pendidikan, Sosial, dan Agama Jurnal Biodjati JURNAL PENDIDIKAN JASMANI DAN OLAHRAGA Chimica et Natura Acta Jurnal Moral Kemasyarakatan JURNAL SAINS INDONESIA Jurnal Kimia Terapan Indonesia Jurnal Pendidikan Kimia (JPKim) Indonesian Journal of Chemistry Jurnal PAJAR (Pendidikan dan Pengajaran) Gelanggang Olahraga: Jurnal Pendidikan Jasmani dan Olahraga (JPJO) Molekul: Jurnal Ilmiah Kimia Jurnal Pedagogik Olahraga Journal of Authentic Research on Mathematics Eduacation (JARME) Jurnal PORKES JUMORA: Jurnal Moderasi Olahraga Jurnal Ilmiah Berkala: Sains dan Terapan Kimia Pharmaceutical Sciences and Research (PSR) Rangkiang Mathematics Journal International Journal of Educational Research Excellence (IJERE) Jurnal Sains dan Teknologi Farmasi Indonesia al Kimiya : Jurnal Ilmu Kimia dan Terapan Journal of Physical Education Health and Sport

Toto Subroto

Department Of Chemistry, Faculty Of Mathematics And Natural Sciences, Universitas Padjadjaran, Jl. Raya Bandung-Sumedang Km 21 Jatinangor, Sumedang, 45363 Jawa Barat, Indonesia

Author-ID : 387376

Agriculture, Biological Sciences & Forestry Arts Humanities Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Computer Science & IT Earth & Planetary Sciences Education Environmental Science Health Professions Immunology & microbiology Languange, Linguistic, Communication & Media Materials Science & Nanotechnology Mathematics Medicine & Pharmacology Public Health Social Sciences Other

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Codon Optimization and Chaperone Assisted Solubilization of Recombinant Human Prethrombin-2 Expressed in Escherichia coli SARONOM SILABAN; IMAN PERMANA MAKSUM; SHABARNI GHAFFAR; KHOMAINI HASAN; SUTARYA ENUS; TOTO SUBROTO; SOETIJOSO SOEMITRO
Microbiology Indonesia Vol. 8 No. 4 (2014): December 2014
Publisher : Indonesian Society for microbiology

Prethrombin-2 (PT2) is a thrombin precursor, which plays a role in the conversion of fibrinogen into fibrin during blood clotting process. Previous study reported that the expression of human prothrombin-2 (rhPT2) in Escherichia coli formed inclusion bodies. The aim of this study was to establish a strategy to express a soluble rhPT2 in E. coli. This study was animed to design and codon optimize human prethrombin-2 gene as well as to optimize the expression condition using four strains of E. coli. The codon adaptation index (CAI) of the unoptimized hpt2 gene was 0.336, with 56.8% GC content. After optimization, the CAI of optimized hpt2 became 1.000 with 53.1% GC content. The optimized gene was successfully cloned into pTWIN1 expression vector. Expression analysis indicated that only E. coli ArcticExpress strain could successfully express a soluble recombinant rhPT2 protein, with only part of rhPT2 being expressed in insoluble form. However, the rest of the E. coli strains used in the experiments failed to express the rhPT2 in soluble form. We are deducing that the success in achieving soluble expression was not only due to the availability of chaperonins Cpn60/Cpn10, which played a crucial role in the protein folding in E. coli ArcticExpress strain, but also due to the codon optimization of hpt2 gene.

Heterologous Expression of Î±-Amylase from Saccharomycopsis fibuligera R64 and its Tyr401Trp Mutant in Pichia pastoris RIEZKI AMALIA; WANGSA TIRTA ISMAYA; FERNITA PUSPASARI; KHOMAINI HASAN; TOTO SUBROTO; DESSY NATALIA; SOETIJOSO SOEMITRO
Microbiology Indonesia Vol. 10 No. 1 (2016): March 2016
Publisher : Indonesian Society for microbiology

Î±-Amylase from Saccharomycopsis fibuligera R64 is a non-adsorbing raw-starch degrading enzyme, a unique characteristic. This character is difficult to explain in the absence of its three-dimensional structure. Here we discuss the expression of a-amylase from Saccharomycopsis fibuligera in Pichia pastoris and the effect of site directed mutagenesis on its activity. A model based on the structure of its homologs suggested mutation of codon of Tyr401 into that of a Trp residue. An activity study using whole cells P. pastoris showed similar substrate degradation rates by cells carrying either the native or mutant amylase encoding gene. However, the purified enzyme of the mutant strain showed faster starch hydrolysis.

Cloning, Expression, and Functional Characterization of Autoactivated Human Prethrombin-2 Synthetic Gene by Using Pichia pastoris SMD1168 As a Host TOTO SUBROTO; WULAN PERTIWI; MUHAMMAD FADHILLAH; KHOMAINI HASAN; OGI BUDIANTORO; SUTARYA ENUS; SOETIJOSO SOEMITRO
Microbiology Indonesia Vol. 10 No. 2 (2016): June 2016
Publisher : Indonesian Society for microbiology

Prethrombin-2 is a thrombin precursor that has important role in blood coagulation. It is the smallest precursor which is activated into thrombin by FXa prior to coagulation process. However, as a commercial theurapetic protein in fibrin sealant component, prethrombin-2 must be activated by ecarin before used. Thus, the production process of this protein needs further purification. In order to eliminate ecarin activation step and to increase production efficiency, we designed, cloned and expressed the recombinant autoactivated human prethrombin-2 in Pichia pastoris SMD1168. The variant was designed with 4 mutations, E40A, D47A, G48P, and E52A, following the result of a previous study. The synthetic variant gene was first optimized to conform with P. pastoris codon preference. The optimized synthetic gene was cloned in pD912 plasmid using XhoI and SacII restriction enzymes. The transformed P. pastoris was selected on agar plate supplemented with 1,000 Âµg.mL-1 Zeocin as a selection marker. This study showed that autoactivated prethrombin-2 was succesfully expressed extracellularly by P. pastoris SMD1168. The activity of recombinant autoactivated prethrombin-2 using a chromogenic substrate S-2238 was 0.540 unit/mg. Taken together, these results demonstrated that autoactivated human prethrombin-2 was successfully produced extracellularly in P. pastoris.

Co-Authors Abdul Alim Kahari, Abdul Alim Abu Bakar M.I. Syihab Alifa I. Nabila Amiruddin, Mohd. Hasril Anita Yuwita Anneke Noviyanti Aprillia, Mutiara Sukma Ari Hardianto Bachti Alisjahbana Basti Andriyoko Bisri Musthofa Budiantoro, Ogi Cindy Aulia Sari D Agus Yusuf Wildan Dani Permana Debby M Sumanti Defi Supriyati, Ayu DESSY NATALIA DESSY NATALIA DESSY NATALIA Dewi Astriany Dhiya Salsabila Dian Budiana, Dian Diana P Rahmawati Didi Suryadi DWI RAHMAWATI Eddy, Diana Rakhmawaty Effendi, Syulastri EKO YULIANTO Elazmanawati Lembong Fadhilah, Muhammad Naufal Fadhlillah, Muhammad Fadillah, Lukky Farihandiandra, Yusron Fathurrohman, Muhammad Andi Fatin, Nuri Nuraini Fauzian Giansyah Febriani Aji Kusuma, Sherlynda FERNITA PUSPASARI Firdaus, Muhammad Lutfi Gifa Nur Arofah Gumelar, Abih Gumilar, Agus Hafizatunnisa, Hafizatunnisa Hambali, Burhan Hamdiyanti, Meti Henry Chandra Hesti Lina Wiraswati Idar Idar Idar Idar Idar Idar Idar, Idar IMAN PERMANA MAKSUM Iman Permana Maksum Iman Permana Maksum Iman Permana Maksum Iman Permana Maksum Iman Permana Maksum Iman Permana Maksum Iman Permana Maksum Iman Permana Maksum Iman Permana Maksum IMAN PERMANA MAKSUM Kharimah, Nurul Ikhsan KHOMAINI HASAN Kusumawardhani, Shinta Laelasari Laelasari Laelasari, Laelasari Leonardus Wiydatmoko Lesmana, Gianjar Mony, Siti Rasnawati Muchamad Subali Noto Muhammad Fadhil Z, Rusdi Noor Rosa (p:43-50) Muhammad Fadhlillah Muhammad Hilman Daniswara Muhammad Yusuf Muhammad Yusuf Muhammad Yusuf Murniaty Simorangkir Murniaty Simorangkir Naufal D, Kevin Nida Yumna Nisa Fauziah Noviyanti, Anneke Nurfairuz, Hasna Siti Nurmalasari, Ratna Nurul Ikhsan Karimah O Suprijana O Suprijana O Suprijana, O OGI BUDIANTORO Ogi Budiantoro Oo Suprijana Opik Taupiqurrohman Opik Taupiqurrohman Pertiwi, Wulan Praja, Ena Suhena Purba Upay Rahmaniar Mulyani Rahmat, Alit Regi Dwi Septian Riezki Amalia Rima Handiyani Rizky Rosjanuardi Rodiana, Ivta Safri Ishmayana Safri Ishmayana Safri Ishmayana Safrl Ismayana Sahroni, Sahroni Santhy Wyantuti Sari Agung, Syennie Sari Syahruni Sari Syahruni SARONOM SILABAN Saronom Silaban Saronom Silaban Saronom Silaban Shabarni Gaffar Shabarni Gaffar Shabarni Gaffar Shabarni Gaffar Shabarni Gaffar Shabarni Gaffar Shabarni Gaffar -, Shabarni Gaffar SHABARNI GHAFFAR Shinta Kusumawardani Siti Soidah Siti Umayah, Siti SOETIJOSO SOEMITRO Soetijoso Soemitro Soetijoso Soemitro Soetijoso Soemitro Soetijoso Soemitro Soetijoso Soemitro Soetijoso Soemitro Sofyan Multazam N Aji Sriwidodo B, Sriwidodo Sriwidodo Sriwidodo Sriwidodo, . Sukma Nuswantara Sukma Nuswantara Sukma Nuswantara Sunan M. Alpiansyah SUTARYA ENUS Sutarya Enus Sutarya Enus Sutarya Enus Syani, Mamay Syfa Nurfadilah Taupiqurrohman, Opik Taupiqurrohman, Opik Tika Pradnjaparamita, Tika Tonah Triana N Meirina Umi Baroroh Unang Supratman Upay, Purba wahyu widayat WANGSA TIRTA ISMAYA Wulan Pertiwi Yani Suryani Yeni W Hartati Yeni Wahyuni Hartati Zakiyyah, Salma Nur

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Journal : Microbiology Indonesia