Application of SUMO fusion technology for the enhancement of stability and activity of lysophospholipase from Pyrococcus abyssi.
Enzyme Stability
Recombinant Fusion Proteins
/ genetics
Escherichia coli
/ genetics
Hydrogen-Ion Concentration
Kinetics
Pyrococcus abyssi
/ genetics
Temperature
Small Ubiquitin-Related Modifier Proteins
/ metabolism
Genetic Vectors
/ metabolism
SUMO-1 Protein
/ genetics
Cloning, Molecular
Solubility
P. abyssi
Biochemical characterization
Enhanced activity
Lysophospholipase
SUMO-fusion
Thermostability
Journal
World journal of microbiology & biotechnology
ISSN: 1573-0972
Titre abrégé: World J Microbiol Biotechnol
Pays: Germany
ID NLM: 9012472
Informations de publication
Date de publication:
09 May 2024
09 May 2024
Historique:
received:
11
02
2024
accepted:
21
04
2024
medline:
9
5
2024
pubmed:
9
5
2024
entrez:
9
5
2024
Statut:
epublish
Résumé
Heterologous production of proteins in Escherichia coli has raised several challenges including soluble production of target proteins, high levels of expression and purification. Fusion tags can serve as the important tools to overcome these challenges. SUMO (small ubiquitin-related modifier) is one of these tags whose fusion to native protein sequence can enhance its solubility and stability. In current research, a simple, efficient and cost-effective method is being discussed for the construction of pET28a-SUMO vector. In order to improve the stability and activity of lysophospholipase from Pyrococcus abyssi (Pa-LPL), a 6xHis-SUMO tag was fused to N-terminal of Pa-LPL by using pET28a-SUMO vector. Recombinant SUMO-fused enzyme (6 H-S-PaLPL) works optimally at 35 °C and pH 6.5 with remarkable thermostability at 35-95 °C. Thermo-inactivation kinetics of 6 H-S-PaLPL were also studied at 35-95 °C with first order rate constant (k
Identifiants
pubmed: 38722449
doi: 10.1007/s11274-024-03998-w
pii: 10.1007/s11274-024-03998-w
doi:
Substances chimiques
Recombinant Fusion Proteins
0
Small Ubiquitin-Related Modifier Proteins
0
SUMO-1 Protein
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
183Informations de copyright
© 2024. The Author(s), under exclusive licence to Springer Nature B.V.
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