Expression of His-tagged NADPH-dependent acetoacetyl-CoA reductase in recombinant Escherichia coli BL-21(DE3).
Affinity chromatography
Enzyme expression
His-tagged
NADPH-dependent acetoacetyl-CoA reductase
Recombinant Escherichia coli
Journal
Journal of bioscience and bioengineering
ISSN: 1347-4421
Titre abrégé: J Biosci Bioeng
Pays: Japan
ID NLM: 100888800
Informations de publication
Date de publication:
Oct 2023
Oct 2023
Historique:
received:
14
04
2023
revised:
14
06
2023
accepted:
01
07
2023
medline:
15
9
2023
pubmed:
28
7
2023
entrez:
27
7
2023
Statut:
ppublish
Résumé
Poly-3-hydroxybutyrate (P(3HB)), a member of the polyhydroxyalkanoate (PHA) family, is a biodegradable polyester with diverse industrial applications. NADPH-dependent acetoacetyl-CoA reductase (phaB) is the enzyme which plays an essential role in P(3HB) synthesis by catalyzing the conversion of the intermediates. The expression of phaB enzyme using the recombinant Escherichia coli BL-21(DE3) and the purification of the synthesized enzyme were studied. The pET-B3 plasmid harbouring the phaB gene derived from Ralstonia eutropha H16, was driven by the lac promoter in E. coli BL-21(DE3). The enzyme was expressed with different induction time, temperatures and cell age. Results showed that the cell age of 4 h, induction time of 12 h at 37°C were identified as the optimal conditions for the enzyme reductase expression. A specific activity of 0.151 U mg
Identifiants
pubmed: 37500302
pii: S1389-1723(23)00228-1
doi: 10.1016/j.jbiosc.2023.07.001
pii:
doi:
Substances chimiques
NADP
53-59-8
acetoacetyl-CoA reductase
EC 1.1.1.36
Hydroxybutyrates
0
Alcohol Oxidoreductases
EC 1.1.-
Polyesters
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
312-319Informations de copyright
Copyright © 2023 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.