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Expression, Purification and Characterization of Recombinant Histidine-tagged L-asparaginase II

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Last updated: 01 Jan 2025

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Abstract

 ESCHERICHIA coli has two L-asparaginase isozymes that have ..... been designated L-asparaginase I and L-asparaginase II. The amino acid sequences of both are rather dissimilar except for a few regions of significant homology. The sequence corresponding to the mature (ansB) was subcloned into pQE-30 expression vector and expressed in E. coli M15. The recombinant histidine-tagged (ansB) was purified to homogeneity by Ni–NTA affinity chromatography and displayed a single 36.0 kDa band on SDS-PAGE. Results revealed that the recombinant His-tagged L-ASNase II was expressed in an active form and its specific activity was estimated to be 286 U/mg. The optimum temperature was attained at 40°C. The enzyme was maximum at pH 7-8. Also the enzyme was stable at 40-50°C. The purified functional enzyme exhibited a specific activity at 286 U/mg and inhibited the growth of human myeloid leukemia cell line (HL-60) with IC50 value of 0.14±0.03 U/ml.

DOI

10.21608/ejbo.2016.2728

Keywords

Anti-tumor, Cytotoxicity, expression, His-tag, L-asparaginase, Leukemia, Purification, Recombinant

Volume

56

Article Issue

3

Related Issue

507

Issue Date

2016-09-01

Receive Date

2016-02-17

Publish Date

2016-09-30

Page Start

707

Page End

722

Print ISSN

0375-9237

Online ISSN

2357-0350

Article File

EJBO_Volume 56_Issue 3_Pages 707-722.pdf

PDF . 383.5kB

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https://ejbo.journals.ekb.eg/article_2728.html

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https://ejbo.journals.ekb.eg/service?article_code=2728

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9

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Original Article

Type Code

111

Publication Type

Journal

Publication Title

Egyptian Journal of Botany

Publication Link

https://ejbo.journals.ekb.eg/

MainTitle

Expression, Purification and Characterization of Recombinant Histidine-tagged L-asparaginase II

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Article

Created At

22 Jan 2023