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l-Asparaginase from Erwinia Chrysanthemi 3937: Cloning, expression and characterization

Paper ID Volume ID Publish Year Pages File Format Full-Text
25516 43578 2007 13 PDF Available
Title
l-Asparaginase from Erwinia Chrysanthemi 3937: Cloning, expression and characterization
Abstract

Bacterial l-asparaginases (l-ASNases) catalyze the conversion of l-asparagine to l-aspartate and ammonia. In the present work, we report the cloning and expression of l-asparaginase from Erwinia chrysanthemi 3937 (Erl-ASNase) in Escherichia coli BL21(DE3)pLysS. The enzyme was purified to homogeneity in a single-step procedure involving cation exchange chromatography on an S-Sepharose FF column. The enzymatic and structural properties of the recombinant enzyme were investigated and the kinetic parameters (Km, kcat) for a number of substrates were determined. In addition, we found that the enzyme can be efficiently immobilized on epoxy-activated Sepharose CL-6B. The immobilized enzyme retains most of its activity (60%) and shows high stability at 4 °C. The approach offers the possibility of designing an Erl-ASNase bioreactor that can be operated over a long period of time with high efficiency, which can be used in leukaemia therapy.

Keywords
ALL, acute lymphoblastic leukaemia; Erl-ASNase, l-asparaginase from Erwinia chrysanthemi; GDH, glutamate dehydrogenase; ORF, open reading frame; PAGE, polyacryamide gel electrophoresis; SDS, sodium dodecyl sulfate; Suc, succinic acidl-Asparaginase; Hydrol
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l-Asparaginase from Erwinia Chrysanthemi 3937: Cloning, expression and characterization
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 127, Issue 4, 20 January 2007, Pages 657–669
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us