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All Journal Jurnal Teknologi Dan Industri Pangan Microbiology Indonesia Alchemy : Journal of Chemistry Agrointek Journal of Food and Life Sciences Jurnal Pangan dan Agroindustri Jurnal Teknologi Pertanian Natural B Indonesian Journal of Human Nutrition agriTECH Majalah Kedokteran Sriwijaya BIOMA : Jurnal Biologi Makassar BIOEDUSCIENCE TEKNOLOGI PANGAN : Media Informasi dan Komunikasi Ilmiah Teknologi Pertanian Indonesian Journal of Halal Research Advances in Food Science, Sustainable Agriculture and Agroindustrial Engineering (AFSSAAE) AJARCDE (Asian Journal of Applied Research for Community Development and Empowerment) ZOOTEC Jurnal Ilmiah Pengabdian Kepada Masyarakat Journal of Research on Community Engagement
Joni Kusnadi
Fakultas Teknologi Pertanian, Universitas Brawijaya, Malang
Religion Agriculture, Biological Sciences & Forestry Humanities Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Computer Science & IT Earth & Planetary Sciences Education Energy Engineering Environmental Science Health Professions Immunology & microbiology Industrial & Manufacturing Engineering Materials Science & Nanotechnology Mechanical Engineering Medicine & Pharmacology Nursing Public Health Social Sciences Veterinary Other

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Journal : Microbiology Indonesia

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Cloning and Gene Expression of AnsZ Encoding L-Asparaginase Enzyme from Local Bacillus sp. RIMA AZARA; IS HELIANTI; JONI KUSNADI; YUNIANTA YUNIANTA
Microbiology Indonesia Vol. 8 No. 2 (2014): June 2014
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (822.691 KB) | DOI: 10.5454/mi.8.2.1

Abstract

L-asparaginase is an enzyme that catalyzes the hydrolysis of L-asparagine into L-aspartic acid and ammonia. In medical aspect, L-asparaginase especially those came from E. coli and Erwinia chrysanthemi used as chemotherapy agent of acute lymphoblastic leukemia (ALL). However, new potential organisms possessing L-asparaginase production capacity with a similar therapeutic effect are still required . In Bacillus subtilis strain 168, there are two kinds of L-asparaginase gene, AnsA and AnsZ. The study of the later L-asparaginase (AnsZ) has not been conducted intensively. The aim of this study is, first, to isolate this gene of L-asparaginase (AnsZ) from local Bacillus sp. and then to express this gene in Escherichia coli. Using PCR-cloning method, an open reading frame (ORF) containing 1128 bp was obtained. The ORF has 99% homology with sequence of L-asparaginase from Bacillus subtilis Bsn5. The gene then was subcloned into pET 21d (+) with his6-tag in the C-terminal of the gene product and expressed in E.coli BL21. L-asparaginase activity analyses showed that recombinant E. coli containing recombinant plasmid with open reading frame (ORF) L-asparaginase (AnsZ) from Bacillus subtilis had higher activity than that is not containing ORF L-asparaginase (AnsZ). Purification with HisPur TM Ni-NTA Purification Kit increased the specific activity of L-asparaginase (AnsZ) enzyme to 29 fold.
Co-Authors A'rasyiJ Imaduddin A.A. Ketut Agung Cahyawan W Adistia Ritri Prihandini Adistia Ritri Prihandini, Adistia Ritri Afifah Puji Hastuti Afifah Puji Hastuti, Afifah Puji Afla Tazakka Aj-juwita Afla Tazakka Aj-juwita Afriliana, Asmak AGUSTIN KRISNA WARDANI Agustin Mardiana, Nur Ahmad Zakiyul Wafi Ainina Ahmad Santoso Ali Masykur Ali Masykur Andayani, Dian Wuri Anik Ardianti Anik Ardianti Atmiral Ernes Ayu Fitriana Wulandari Baiq Dina Mariana Benediktus Hendra Perkasa Citra Firdhausi Citra Firdhausi Daimon Syukri Dedi Dedi Denta, Anggeria Oktavisa Dian Widya Ningtyas Dila Rahmawati Dr.Ir. Yunianta, DEA Dwi Esti Prayanti Dwi Ratnaningsih DWI SURYANTO Edi Priyo Utomo Elke Galuh Primurdia Elke Galuh Primurdia Elok Zubaedah Elok Zubaidah Endrika Widyastuti Endrika Widyastuti Eni Purwanti Eni Purwanti, Eni Erni Sofia Muritni Erni Sofia Murtini Erryana Martati Estri Laras Arumingtyas Estri Laras Arumingtyas Eunike Debora Purba Fadri, Rince Alfia Faiqotur Rohmania Feronika Heppy Sriherfyna Fitri Nurzanah Fitriyana, Nurul Isnaini Freini Dessi Efendi Hakiki, Hilda Maghfirotu Hamidin Rasulu Hana Susanti Maleta Hannes Dwi Novia Hariry, Amelia Hasria Alang Herlambang, Dwi Hernandi, Kevin Hohn Hilda Maghfirotu Hakiki Ima Andriastuti IS HELIANTI Ito, Nobutaka Jamhari Jamhari Jaya Mahar Maligan Kartika Cahyania Pratiwi Kevin Hohn Hernandi Khairul Ambri Khotibul Umam Al-Awwaly Khotibul Umam Al-Awwaly Khusnul Khotimah Khusnul Khotimah KIKI FIBRIANTO Lenny Yudha Febriyanti Lenny Yudha Febriyanti Lia Ratnawati Lilis Karlina Malsa Fian Yana Malsa Fian Yana Mega Kristanti Ayuratri Muhammad Reza Ramadhana Muhammad Reza Ramadhana, Muhammad Reza Nazir, N Nenu Maria Ovy Sonia Nenu Maria Ovy Sonia Nisa Harmia Yuliasih Noor Hidayat, Noor Novi Vensia Magdalena Novi Vensia Magdalena Novita Wijayanti Nur Istianah Nur Istianah Nur Kusmiyati Pamulanti, Anjar Pebiningrum, Arlinda Pendik Setiawan Pratiwi Anggun Retnowati Pratiwi Anggun Retnowati Putri, Annisyia Zarina Raida Amelia Ifadah Raida Amelia Ifadah Rif'ah, Hamidah Itha'atur RIMA AZARA Rinelda Ayu Sintasari Rinelda Ayu Sintasari Rukmi, Widya Dwi Septi, Nur Diana Shanty Karinawatie Sudarma Dita Wijayanti Sudarma Dita Wijayanti Sudarminto Setyo Yuwono Suharjono Suharjono Suharjono Suharjono, Suharjono Sutik Rahayu Endah Sutrisno Adi Prayitno Suwasono, Sony Tabitha Larasati Tabitha Larasati Tanti Untari Tanti Untari Teti Estiasih Titis Sari Kusuma Tri Ardyati Tri Ardyati Try Ardyati Tutik Murniasih Tuty Anggraini Vivi Retno Sari Vivi Retno Sari Widya Dwi Rukmi Putri Yan Abdi Nugroho Yan Abdi Nugroho Yohana Lydia Rissa Tanjung Yohana Lydia Rissa Tanjung Zuliyaningsih Zuliyaningsih