Enoate Reductases of Clostridia

Abstract

The enr genes specifying enoate reductases of Clostridium tyrobutyricum and Clostridium thermoaceticum were cloned and sequenced. Sequence comparison shows that enoate reductases are similar to a family of flavoproteins comprising 2,4-dienoyl-coenzyme A reductase from Escherichia coli and old yellow enzyme from yeast. The C. thermoaceticum enr gene product was expressed in recombinant Escherichia coli cells growing under anaerobic conditions. The recombinant enzyme was purified and characterized.

Alkene hydrogenation activity of enoate reductases for an environmentally benign biosynthesis of adipic acid

“…34 Compared with OYEs, ERs are less characterized, because these enzymes were found to be oxygen-sensitive. 35–37 Clostridial ERs use NADH as an electron donor and contain FAD, FMN, and an [4Fe–4S] iron–sulfur cluster. 37,38 ERs can reduce CC double bonds in a variety of monoesters and monoacids, including non-activated 2-enoates, a reaction not catalyzed by OYEs.…”

“…35–37 Clostridial ERs use NADH as an electron donor and contain FAD, FMN, and an [4Fe–4S] iron–sulfur cluster. 37,38 ERs can reduce CC double bonds in a variety of monoesters and monoacids, including non-activated 2-enoates, a reaction not catalyzed by OYEs. 36,39 Although both OYEs and ERs have the potential to catalyze the hydrogenation of unsaturated six-carbon dicarboxylic acids to adipic acid, these studies have not yet been reported.…”

Biosynthesis of Mupirocin by Pseudomonas fluorescens NCIMB 10586 Involves Parallel Pathways

“…which shows high homology to dienoyl CoA reductases. 20 When the ∆mupF strain was examined (Fig. 4B(c)), the metabolite profile was almost identical to that from ∆mupC with the addition of the previously reported 16 mupirocin F1 18a containing a 6--hydroxy, 7--keto moiety (along with 50% of its 8--epimer 18b) and its epoxide--mediated rearrangement (see Scheme S1) product mupi--rocin F2 19 as additional major (ca 8 mgL --1 ) metabolites.…”

‘New uses for an Old Enzyme’ – the Old Yellow Enzyme family of flavoenzymes

“…A barrel domain related to OYE forms part of trimethylamine dehydrogenase (Lim et al, 1986), and is used as a module in several other multidomain proteins (Scrutton, 1994), such as E. coli 2,4-dienoyl-CoA-reductase (He et al, 1997) and enoate reductases of clostridia (Rohdich et al, 2001). Whilst some of the putative active site residues differ in these distant relatives of OYE, there is a strong sequence conservation in a core region of around 40 amino acids.…”