| dc.contributor.author |
Kotzia, GA |
en |
| dc.contributor.author |
Labrou, NE |
en |
| dc.date.accessioned |
2014-06-06T06:47:52Z |
|
| dc.date.available |
2014-06-06T06:47:52Z |
|
| dc.date.issued |
2007 |
en |
| dc.identifier.issn |
01681656 |
en |
| dc.identifier.uri |
http://dx.doi.org/10.1016/j.jbiotec.2006.07.037 |
en |
| dc.identifier.uri |
http://62.217.125.90/xmlui/handle/123456789/3828 |
|
| dc.subject |
Enzyme immobilization |
en |
| dc.subject |
Hydrolase |
en |
| dc.subject |
l-Asparaginase |
en |
| dc.subject |
Leukaemia |
en |
| dc.subject.other |
Bioreactors |
en |
| dc.subject.other |
Cloning |
en |
| dc.subject.other |
Escherichia coli |
en |
| dc.subject.other |
Genes |
en |
| dc.subject.other |
Ion exchange |
en |
| dc.subject.other |
Reaction kinetics |
en |
| dc.subject.other |
Hydrolase |
en |
| dc.subject.other |
L-Asparaginase |
en |
| dc.subject.other |
Leukaemia |
en |
| dc.subject.other |
Enzymes |
en |
| dc.subject.other |
ammonia |
en |
| dc.subject.other |
asparaginase |
en |
| dc.subject.other |
asparagine |
en |
| dc.subject.other |
aspartic acid |
en |
| dc.subject.other |
recombinant enzyme |
en |
| dc.subject.other |
sepharose |
en |
| dc.subject.other |
amino terminal sequence |
en |
| dc.subject.other |
article |
en |
| dc.subject.other |
bioreactor |
en |
| dc.subject.other |
enzyme activity |
en |
| dc.subject.other |
enzyme analysis |
en |
| dc.subject.other |
enzyme immobilization |
en |
| dc.subject.other |
enzyme kinetics |
en |
| dc.subject.other |
enzyme purification |
en |
| dc.subject.other |
enzyme stability |
en |
| dc.subject.other |
enzyme structure |
en |
| dc.subject.other |
Escherichia coli |
en |
| dc.subject.other |
ion exchange chromatography |
en |
| dc.subject.other |
leukemia |
en |
| dc.subject.other |
molecular cloning |
en |
| dc.subject.other |
nonhuman |
en |
| dc.subject.other |
nucleotide sequence |
en |
| dc.subject.other |
Pectobacterium chrysanthemi |
en |
| dc.subject.other |
priority journal |
en |
| dc.subject.other |
protein expression |
en |
| dc.subject.other |
sequence analysis |
en |
| dc.subject.other |
thermostability |
en |
| dc.subject.other |
Amino Acid Sequence |
en |
| dc.subject.other |
Antineoplastic Agents |
en |
| dc.subject.other |
Asparaginase |
en |
| dc.subject.other |
Bacterial Proteins |
en |
| dc.subject.other |
Cloning, Molecular |
en |
| dc.subject.other |
Escherichia coli |
en |
| dc.subject.other |
Heat |
en |
| dc.subject.other |
Kinetics |
en |
| dc.subject.other |
Models, Molecular |
en |
| dc.subject.other |
Molecular Sequence Data |
en |
| dc.subject.other |
Pectobacterium chrysanthemi |
en |
| dc.subject.other |
Sequence Alignment |
en |
| dc.subject.other |
Stereoisomerism |
en |
| dc.subject.other |
Bacteria (microorganisms) |
en |
| dc.subject.other |
Erwinia chrysanthemi str. 3937 |
en |
| dc.subject.other |
Escherichia coli |
en |
| dc.title |
l-Asparaginase from Erwinia Chrysanthemi 3937: Cloning, expression and characterization |
en |
| heal.type |
journalArticle |
en |
| heal.identifier.primary |
10.1016/j.jbiotec.2006.07.037 |
en |
| heal.publicationDate |
2007 |
en |
| heal.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. © 2006 Elsevier B.V. All rights reserved. |
en |
| heal.journalName |
Journal of Biotechnology |
en |
| dc.identifier.issue |
4 |
en |
| dc.identifier.volume |
127 |
en |
| dc.identifier.doi |
10.1016/j.jbiotec.2006.07.037 |
en |
| dc.identifier.spage |
657 |
en |
| dc.identifier.epage |
669 |
en |