Purification and partial characterization of L-asparaginase enzyme produced by newly isolated Bacillus sp

Purification and partial characterization of L-asparaginase enzyme produced by newly isolated Bacillus sp

A new bacterial producing L-asparaginase was successfully isolated from Sungai Klah Hot Spring, Perak, Malaysia and identified as Bacillus sp. It was the best L-asparaginase producer as compared to other isolates. Production of L-asparaginase from the microbial strain was carried out under liquid fe...

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Main Authors: Jimat, Dzun Noraini, Mohamed, Intan Baizura Firda, Azmi, Azlin Suhaida, Jamal, Parveen
Format: Article
Language:English
English
Published: Kulliyah of Engineering, International Islamic University Malaysia (IIUM) 2017
Subjects:
QR Microbiology
TP248.13 Biotechnology
Online Access:http://irep.iium.edu.my/59636/
http://irep.iium.edu.my/59636/
http://irep.iium.edu.my/59636/1/PURIFICATION%20AND%20PARTIAL%20CHARACTERIZATION%20OF%20L-ASPARAGINASE%20ENZYME%20PRODUCED%20BY%20NEWLY%20ISOLATED%20BACILLUS%20sp.pdf
http://irep.iium.edu.my/59636/7/Purification%20and%20partial%20characterization%20of%20L-asparaginase%20enzyme%20produced%20by%20newly%20isolated%20Bacillus%20Sp..pdf
id iium-59636
recordtype eprints
spelling iium-596362018-03-20T05:16:16Z http://irep.iium.edu.my/59636/ Purification and partial characterization of L-asparaginase enzyme produced by newly isolated Bacillus sp Jimat, Dzun Noraini Mohamed, Intan Baizura Firda Azmi, Azlin Suhaida Jamal, Parveen QR Microbiology TP248.13 Biotechnology A new bacterial producing L-asparaginase was successfully isolated from Sungai Klah Hot Spring, Perak, Malaysia and identified as Bacillus sp. It was the best L-asparaginase producer as compared to other isolates. Production of L-asparaginase from the microbial strain was carried out under liquid fermentation. The crude enzyme was then centrifuged and precipitated with ammonium sulfate before being further purified by chromatographic method. HiTrap DEAE-Sepharose Fast Flow ion exchange chromatography followed by separation on Superose 12 gel filtration were used to obtain pure enzyme. The purified enzyme showed 10.11 U/mg of specific activity, 50.07% yield with 2.21 fold purification. The purified enzyme was found to be dimer in form, with a molecular weight of 65 kDa as estimated by SDS-PAGE. The maximum activity of the purified L-asparaginase was observed at pH 9 and temperature of 60 oC. Kajian penyelidikan ini telah berjaya menghasilkan enzim L-aspraginase daripada bakteria-bakteria baru yang diambil dari Kolam Air Panas, Sungai Klah, Perak, Malaysia dan ia dikenal pasti sebagai Bacillus sp. Teknik isolasi bakteria ini adalah teknik terbaik dalam menghasilkan enzim L-asparaginase berbanding teknik-teknik lain. Penghasilan enzim ini dibuat menerusi proses fermentasi. Kultur bakteria yang diperolehi diempar dan diikuti presipitasi menggunakan ammonium sulfat sebelum proses penulenan seterusnya dilakukan menggunakan kaedah kromatografi. Kolum penukaran ion jenis HiTrap DEAE-Sepharose Fast Flow diikuti dengan pemisahan oleh gel Superose 12 telah digunakan untuk mendapatkan enzim L-asparaginase yang tulen. Hasil kajian mendapati enzim tulen yang diperolehi mempunyai aktiviti spesifik sebanyak 10.11 U/mg daripada 50.07% enzim yang dihasilkan dan berjaya mencapai 2.21 kali ganda tulen dari enzim tanpa proses penulenan. Enzim tulen yang diperolehi didapati dalam bentuk “dimer“ dengan berat molekular sebanyak 65 kDa yang ditentukan menerusi SDS-PAGE. Enzim ini juga menunjukkan aktiviti yang tinggi pada pH 9 dan suhu 60 oC. Kulliyah of Engineering, International Islamic University Malaysia (IIUM) 2017-12 Article PeerReviewed application/pdf en http://irep.iium.edu.my/59636/1/PURIFICATION%20AND%20PARTIAL%20CHARACTERIZATION%20OF%20L-ASPARAGINASE%20ENZYME%20PRODUCED%20BY%20NEWLY%20ISOLATED%20BACILLUS%20sp.pdf application/pdf en http://irep.iium.edu.my/59636/7/Purification%20and%20partial%20characterization%20of%20L-asparaginase%20enzyme%20produced%20by%20newly%20isolated%20Bacillus%20Sp..pdf Jimat, Dzun Noraini and Mohamed, Intan Baizura Firda and Azmi, Azlin Suhaida and Jamal, Parveen (2017) Purification and partial characterization of L-asparaginase enzyme produced by newly isolated Bacillus sp. IIUM Engineering Journal, 18 (2). ISSN 1511-788X E-ISSN 2289-7860 http://journals.iium.edu.my/ejournal/index.php/iiumej/pages/view/future_articles
repository_type Digital Repository
institution_category Local University
institution International Islamic University Malaysia
building IIUM Repository
collection Online Access
language English
English
topic QR Microbiology
TP248.13 Biotechnology
spellingShingle QR Microbiology
TP248.13 Biotechnology
Jimat, Dzun Noraini
Mohamed, Intan Baizura Firda
Azmi, Azlin Suhaida
Jamal, Parveen
Purification and partial characterization of L-asparaginase enzyme produced by newly isolated Bacillus sp
description A new bacterial producing L-asparaginase was successfully isolated from Sungai Klah Hot Spring, Perak, Malaysia and identified as Bacillus sp. It was the best L-asparaginase producer as compared to other isolates. Production of L-asparaginase from the microbial strain was carried out under liquid fermentation. The crude enzyme was then centrifuged and precipitated with ammonium sulfate before being further purified by chromatographic method. HiTrap DEAE-Sepharose Fast Flow ion exchange chromatography followed by separation on Superose 12 gel filtration were used to obtain pure enzyme. The purified enzyme showed 10.11 U/mg of specific activity, 50.07% yield with 2.21 fold purification. The purified enzyme was found to be dimer in form, with a molecular weight of 65 kDa as estimated by SDS-PAGE. The maximum activity of the purified L-asparaginase was observed at pH 9 and temperature of 60 oC. Kajian penyelidikan ini telah berjaya menghasilkan enzim L-aspraginase daripada bakteria-bakteria baru yang diambil dari Kolam Air Panas, Sungai Klah, Perak, Malaysia dan ia dikenal pasti sebagai Bacillus sp. Teknik isolasi bakteria ini adalah teknik terbaik dalam menghasilkan enzim L-asparaginase berbanding teknik-teknik lain. Penghasilan enzim ini dibuat menerusi proses fermentasi. Kultur bakteria yang diperolehi diempar dan diikuti presipitasi menggunakan ammonium sulfat sebelum proses penulenan seterusnya dilakukan menggunakan kaedah kromatografi. Kolum penukaran ion jenis HiTrap DEAE-Sepharose Fast Flow diikuti dengan pemisahan oleh gel Superose 12 telah digunakan untuk mendapatkan enzim L-asparaginase yang tulen. Hasil kajian mendapati enzim tulen yang diperolehi mempunyai aktiviti spesifik sebanyak 10.11 U/mg daripada 50.07% enzim yang dihasilkan dan berjaya mencapai 2.21 kali ganda tulen dari enzim tanpa proses penulenan. Enzim tulen yang diperolehi didapati dalam bentuk “dimer“ dengan berat molekular sebanyak 65 kDa yang ditentukan menerusi SDS-PAGE. Enzim ini juga menunjukkan aktiviti yang tinggi pada pH 9 dan suhu 60 oC.
format Article
author Jimat, Dzun Noraini
Mohamed, Intan Baizura Firda
Azmi, Azlin Suhaida
Jamal, Parveen
author_facet Jimat, Dzun Noraini
Mohamed, Intan Baizura Firda
Azmi, Azlin Suhaida
Jamal, Parveen
author_sort Jimat, Dzun Noraini
title Purification and partial characterization of L-asparaginase enzyme produced by newly isolated Bacillus sp
title_short Purification and partial characterization of L-asparaginase enzyme produced by newly isolated Bacillus sp
title_full Purification and partial characterization of L-asparaginase enzyme produced by newly isolated Bacillus sp
title_fullStr Purification and partial characterization of L-asparaginase enzyme produced by newly isolated Bacillus sp
title_full_unstemmed Purification and partial characterization of L-asparaginase enzyme produced by newly isolated Bacillus sp
title_sort purification and partial characterization of l-asparaginase enzyme produced by newly isolated bacillus sp
publisher Kulliyah of Engineering, International Islamic University Malaysia (IIUM)
publishDate 2017
url http://irep.iium.edu.my/59636/
http://irep.iium.edu.my/59636/
http://irep.iium.edu.my/59636/1/PURIFICATION%20AND%20PARTIAL%20CHARACTERIZATION%20OF%20L-ASPARAGINASE%20ENZYME%20PRODUCED%20BY%20NEWLY%20ISOLATED%20BACILLUS%20sp.pdf
http://irep.iium.edu.my/59636/7/Purification%20and%20partial%20characterization%20of%20L-asparaginase%20enzyme%20produced%20by%20newly%20isolated%20Bacillus%20Sp..pdf
first_indexed 2023-09-18T21:24:30Z
last_indexed 2023-09-18T21:24:30Z
_version_ 1777412100709154816

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