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RIMA AZARA
Universitas Muhammadiyah Sidoarjo
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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 Adawiyah, Ro’biatul Ahmad Syauqi Firdaus Al Machfudz Amarulloh, Rochma Andriani Eko Prihatiningrum, Andriani Eko Anggita Dhea Novita Arvialika, Yesy Dwita Atik Wahyuni Budiandari , Rahmah Utami Desi Utami Desi DWI SURYANTO Eka Sabela Elshiva, Danya Mozza Erika Puspita Sari Faiza Nur Aini Frida Dwi Aggraeni Hadiah Fitriyah Hikmah, Allysa Rowihatunnufus Ida Agustini Saidi Indah Apriliana Sari W. Indahsari, Fitriyah Nur IS HELIANTI Jian Permani Ningsih Joni Kusnadi Kinanti, Kintan Sari Laksana, Cakra Lesmana, Mochammad Affan Lukman Hudi M. Vikri Baihaggi Vikri Maziya, Zidny Misbachul Ulum NICO ARDIAWAN PUTRA NICO Nur Aziza Nur Ravita Hanun Nurbaya, Syaraifa Ramadhani Oktarina, Pramitha Wahyu Poppy Diana Sari Poppy Diana Sari, Poppy Diana Puspita Handayani Rafdiansyah, Muhammad Ainur Rahmah Utami Budiandari Ramadhani Nurbaya, Syarifa Robiatul Adawiyah Rochman, Moch Fatchur Sabela, Eka Sari, Erika Puspita Selviani Selviani Sutiono Sutiono Syarifa Ramadhani Nurbaya WDP, Al Machfud Widayanti, Silvi Zahiroh, Alza Della Zakiyah, Mei Dita Fitrotus