Extraction and purification of L-Asparaginase II from local isolate of Proteus vulgaris

Forty one isolates of genus Proteus were collected from 140 clinical specimens such as urine, stool, wound, burn, and ear swabs from patients of both sex. These isolates were identified to three Proteus spp. P. mirabilis, P. vulgaris and P. penneri .The ability of these bacteria to produce L-aspa...

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Main Author: Baghdad Science Journal
Format: Article
Language:English
Published: University of Baghdad, College of Science for Women 2011-03-01
Series:مجلة بغداد للعلوم
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Online Access:http://bsj.uobaghdad.edu.iq/index.php/BSJ/article/view/2524
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author Baghdad Science Journal
author_facet Baghdad Science Journal
author_sort Baghdad Science Journal
collection DOAJ
description Forty one isolates of genus Proteus were collected from 140 clinical specimens such as urine, stool, wound, burn, and ear swabs from patients of both sex. These isolates were identified to three Proteus spp. P. mirabilis, P. vulgaris and P. penneri .The ability of these bacteria to produce L-asparaginase II by using semi quantitative and quantitative methods was determined. P. vulgaris Pv.U.92 was distinguished for high level of L-asparaginase II production with specific activity 1.97 U/mg. Optimum conditions for enzyme production were determined; D medium with 0.3% of L-asparagine at pH 7.5 with temperature degree 35°C for incubation. Ultrasonication was used to destroy the P. vulgaris Pv.U.92 cells then ASNase II was extracted and purified throughout several purification steps including precipitation with (NH4)2SO4(60-80%), DEAE-cellulose ion exchanger chromatography followed by Sephacryl S-300 filtration. The specific activity was 155.6 U/ mg and the purification fold was 27.3 with 10.4% yield.
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spelling doaj-art-2410b8ad81a54aeaa96be6d6523ee33f2025-08-20T03:33:14ZengUniversity of Baghdad, College of Science for Womenمجلة بغداد للعلوم2078-86652411-79862011-03-018110.21123/bsj.8.1.509-518Extraction and purification of L-Asparaginase II from local isolate of Proteus vulgarisBaghdad Science JournalForty one isolates of genus Proteus were collected from 140 clinical specimens such as urine, stool, wound, burn, and ear swabs from patients of both sex. These isolates were identified to three Proteus spp. P. mirabilis, P. vulgaris and P. penneri .The ability of these bacteria to produce L-asparaginase II by using semi quantitative and quantitative methods was determined. P. vulgaris Pv.U.92 was distinguished for high level of L-asparaginase II production with specific activity 1.97 U/mg. Optimum conditions for enzyme production were determined; D medium with 0.3% of L-asparagine at pH 7.5 with temperature degree 35°C for incubation. Ultrasonication was used to destroy the P. vulgaris Pv.U.92 cells then ASNase II was extracted and purified throughout several purification steps including precipitation with (NH4)2SO4(60-80%), DEAE-cellulose ion exchanger chromatography followed by Sephacryl S-300 filtration. The specific activity was 155.6 U/ mg and the purification fold was 27.3 with 10.4% yield.http://bsj.uobaghdad.edu.iq/index.php/BSJ/article/view/2524L-Asparaginase II, P. vulgaris, DEAE-cellulose, Sephacryl S-300.
spellingShingle Baghdad Science Journal
Extraction and purification of L-Asparaginase II from local isolate of Proteus vulgaris
مجلة بغداد للعلوم
L-Asparaginase II, P. vulgaris, DEAE-cellulose, Sephacryl S-300.
title Extraction and purification of L-Asparaginase II from local isolate of Proteus vulgaris
title_full Extraction and purification of L-Asparaginase II from local isolate of Proteus vulgaris
title_fullStr Extraction and purification of L-Asparaginase II from local isolate of Proteus vulgaris
title_full_unstemmed Extraction and purification of L-Asparaginase II from local isolate of Proteus vulgaris
title_short Extraction and purification of L-Asparaginase II from local isolate of Proteus vulgaris
title_sort extraction and purification of l asparaginase ii from local isolate of proteus vulgaris
topic L-Asparaginase II, P. vulgaris, DEAE-cellulose, Sephacryl S-300.
url http://bsj.uobaghdad.edu.iq/index.php/BSJ/article/view/2524
work_keys_str_mv AT baghdadsciencejournal extractionandpurificationoflasparaginaseiifromlocalisolateofproteusvulgaris