Radical-based photoinactivation of fatty acid photodecarboxylases.
Decarboxylation
Fatty acid photodecarboxylase
Flavoprotein
Photoinactivation
Journal
Analytical biochemistry
ISSN: 1096-0309
Titre abrégé: Anal Biochem
Pays: United States
ID NLM: 0370535
Informations de publication
Date de publication:
01 07 2020
01 07 2020
Historique:
received:
31
03
2020
revised:
16
04
2020
accepted:
17
04
2020
pubmed:
30
4
2020
medline:
20
1
2021
entrez:
30
4
2020
Statut:
ppublish
Résumé
Fatty acid photodecarboxylases (FAP) are a recently discovered family of FAD-containing, light-activated enzymes, which convert fatty acids to n-alkanes/alkenes with potential applications in the manufacture of fine and speciality chemicals and fuels. Poor catalytic stability of FAPs is however a major limitation. Here, we describe a methodology to purify catalytically stable and homogeneous samples of recombinant Chlorella variabilis NC64A FAP (CvFAP) from Escherichia coli. We demonstrate however that blue light-exposure, which is required for photodecarboxylase activity, also leads to irreversible inactivation of the enzyme, especially in the absence of palmitate substrate. Photoinactivation is attributed to formation of protein based organic radicals, which were observed by EPR spectroscopy. To suppress photoinactivation, we prepared stable and catalytically active FAP in the dark. The steady-state kinetic parameters of CvFAP (k
Identifiants
pubmed: 32348726
pii: S0003-2697(20)30281-5
doi: 10.1016/j.ab.2020.113749
pii:
doi:
Substances chimiques
Fatty Acids
0
Free Radicals
0
Carboxy-Lyases
EC 4.1.1.-
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
113749Subventions
Organisme : Biotechnology and Biological Sciences Research Council
ID : BB/N013980/1
Pays : United Kingdom
Informations de copyright
Copyright © 2020 The Author(s). Published by Elsevier Inc. All rights reserved.