TABLE 1.

Purification of the caffeyl-CoA reductase-Etf complex from A. woodii

Purification step	Protein	Specific activitya	Yield	Purification
	mg	units/mg	%	-fold
Cell-free extract	2579.2	0.57	100.0	1.0
Cytoplasm	2161.9	0.57	83.2	1.0
Q Sepharose	381.4	2.51	65.0	4.4
Phenyl-Sepharose	99.5	7.83	52.9	13.7
Blue Sepharose	46.5	12.93	40.8	22.6
Gel filtration Sephacryl S-300	14.5	13.61	13.4	23.8

^a The activity of the caffeyl-CoA reductase-Etf complex was determined by following the caffeyl-CoA-dependent oxidation of reduced methyl viologen at 604 nm. The assay buffer (buffer B) contained 5 mm reduced methyl viologen (reduced with 1 mm sodium dithionite) as electron donor, 5 μm caffeyl-CoA, and the caffeyl-CoA regeneration system (200 units of the CoA transferase CarA and 0.25 mm caffeate) under an atmosphere of 100% N₂. Measurements were performed at 30 °C in anaerobic cuvettes.