NADH dehydrogenase, purification

NADH dehydrogenase and, 189 ubiquinone reductase and, 178,182,183 Cholesterol, side chain cleavage, 83, 84-85 Choline, oxidation to betaine, 260 Choline dehydrogenase electron transport system and, 261-263 properties, 260-201 Chromatium sulfate reduction by, 281 transhydrogenase of, 54 function of, 80 molecular properties, 58, 69 purification, 55, 56 Chromium... 

Enzyme Preparation. This enzyme from rabbit muscle is available commercially (Boehringer Manheim Corp.) or may be prepared by the method of Tietz and Ochoa. Further purification may be achieved by the technique of Cottam et al.° Pyruvate kinase activity is conveniently assayed by coupling the reduction of pyruvate, produced from phos-phoenolpyruvate, to lactate, catalyzed by lactate dehydrogenase. The NADH consumed in this reaction is assayed spectrophotometrically. Studies of reversible inhibition are carried out directly in the reaction cuvette, with appropriate controls to eliminate the possibility that inhibition is due to inactivation by DMPA of the lactate dehydrogenase or other assay reagents. 

Weianann C (1915) Improvements in bacterial fermentation of carbohydrates and in bacterial cultures for the same. British Patent 4845 Welch RW (1991) Purification and studies of two butanol (ethanol) dehydrogenases and the effects of rifampicin and chloramphenicol on other enzymes important in the production of butyrate and butanol in Clostridium acetobutylicum ATCC 824. PhD thesis. Rice University, Houston TX Welch RW, Rudolph FB, Papoutsakis ET (1989) Purification and characterization of the NADH-dependent butanol dehydrogenase from Clostridium acetobutylicum (ATCC 824). Arch Biochem Biophys 273 309-318... 

Camp PJ, Randall DD. Purification and characterization of the pea chloroplast pyruvate dehydrogenase complex. A source of acetyl-CoA and NADH for fatty acid biosynthesis. Plant Physiol 1985 77 571-577. 

We have already described the purification of a chloroplastic NADP-malate dehydrogenase (Ferte et al., 1982). It is a 56 000 daltons dimeric protein made up of two apparently identical subunits. This enzyme plays an important role in the generation of reducing power used in the reduction cytoplasmic reactions. NADPH from photosynthesis reduces oxaloacetate to malate in the stroma. This reaction is catalyzed by NADP-MDH. A malate translocator exports this acid outside the chloroplast where NAD-MDH generates NADH. 

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