Pseudomonas aeruginosa recombinant L-asparaginase: Large scale production, purification, and cytotoxicity on THP-1, MDA-MB-231, A549, Caco2 and HCT-116 cell lines.
Acute lymphoblastic leukemia
Cloning
Cytotoxicity
Expression
l-asparaginase
Journal
Protein expression and purification
ISSN: 1096-0279
Titre abrégé: Protein Expr Purif
Pays: United States
ID NLM: 9101496
Informations de publication
Date de publication:
05 2021
05 2021
Historique:
received:
07
09
2020
revised:
02
12
2020
accepted:
03
01
2021
pubmed:
14
1
2021
medline:
25
12
2021
entrez:
13
1
2021
Statut:
ppublish
Résumé
In previous studies Pseudomonas aeruginosal-ASNase complete coding sequence gene, 984 bp (GenBank accession number KU161101.2) was isolated by PCR, cloned into pET28a(+) vector, expressed in E. coli DE3(BL21) pLysS, purified to apparent homogeneity and biochemically characterized. In the present work we highlight large scale production, affinity purification of the recombinant enzyme, effect of osmolytes on the stability of the l-ASNase and cytotoxicity on different cancer cell lines. Successful overexpression was achieved in E. coli as a 6-His-Tag fusion protein after 18 h of induction with lactose at a concentration of 2 g/L in fermentation medium and at 37 °C. The recombinant enzyme was purified to homogeneity using Ni
Identifiants
pubmed: 33440252
pii: S1046-5928(21)00003-6
doi: 10.1016/j.pep.2021.105820
pii:
doi:
Substances chimiques
Antineoplastic Agents
0
Bacterial Proteins
0
Recombinant Proteins
0
Asparaginase
EC 3.5.1.1
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
105820Informations de copyright
Copyright © 2021 Elsevier Inc. All rights reserved.