@inbook{8de25779e0734ad3b2cf48b5df6e0a77,
title = "Heterologous production, reconstitution and EPR spectroscopic analysis of prFMN dependent enzymes",
abstract = "The recent discovery of the prenylated FMN (prFMN) cofactor has led to a renewed interest in the prFMN-dependent UbiD family of enzymes. The latter catalyses the reversible decarboxylation of alpha-beta unsaturated carboxylic acids and features widely in microbial metabolism. The flavin prenyltransferase UbiX synthesizes prFMN from reduced FMN and phosphorylated dimethylallyl precursors. Oxidative maturation of the resulting prFMNreduced species to the active prFMNiminium form is required for UbiD activity. Heterologous production of active holo-UbiD requires co-expression of UbiX, but the levels of prFMN incorporation and oxidative maturation appear variable. Detailed protocols and strategies for in vitro reconstitution and oxidative maturation of UbiD are presented that can yield an alternative source of active holo-UbiD for biochemical studies.",
keywords = "Crystallography, Decarboxylase, EPR, Iminium, Prenyltransferase, prFMN, Radical, UbiD, UbiX",
author = "Marshall, \{Stephen A.\} and Payne, \{Karl A.P.\} and Karl Fisher and Deepankar Gahloth and Bailey, \{Samuel S.\} and Arune Balaikaite and Annica Saaret and Irina Gostimskaya and Godwin Aleku and Huanming Huang and Rigby, \{Stephen E.J.\} and David Procter and David Leys",
note = "Publisher Copyright: {\textcopyright} 2019 Elsevier Inc.",
year = "2019",
doi = "10.1016/bs.mie.2019.03.022",
language = "English",
isbn = "9780128168295",
series = "Methods in Enzymology",
publisher = "ACADEMIC PRESS INC",
pages = "489--508",
editor = "Palfey, \{Bruce A.\}",
booktitle = "New Approaches for Flavin Catalysis",
address = "United States",
}