Dehydrogenases, purification

Umezawa T, Davin LB, Lewis NG (1991) Formation of lignans (-)-secoisolariciresinol and (-)-matairesinol with Forsythia intermedia ceU-free extracts. J Biol Chem 266 10210-10217 Okunishi T, Sakakibara N, Suzuki S et al (2004) Stereochemistry of matairesinol formation by Daphne secoisolariciresinol dehydrogenase. J Wood Sci 50 77-81 XiaZ-Q, Costa MA, Pelissier HC et al (2001) Secoisolariciresinol dehydrogenase purification, cloning, and functional expression. J Biol Chem 276 12614-12623... 

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Xia, Z.Q., Costa, M.A., Pelissier, H.C., Davin, L.B. and Lewis, N.G. (2001) Secoisolari-ciresinol dehydrogenase purification, cloning, and functional expression. Implications for human health protection. /. Biol. Chem., 276, 12614-23. 

Dehydrogenases. One recent development in affinity chromatography is that of the use of group specific supports, already mentioned briefly. The use of group specific supports has found particular application in dehydrogenase purification with affinity columns composed of immobilised nucleotides. 

Lange LH, Sytkowski J, Vallee BL (1976) Human liver alcohol dehydrogenase purification, composition, and catalytic features. Biochemistry 15 4687-4693... 

Ligand affinity chromatography of succinate thiokinase Purification of glyceraldehyde 3-phosphate dehydrogenase Purification of glycogen phosphorylase... 

AflSnity chromatographic purification 192 of adenosine triphosphatase from rat liver mitochondria Affinity chromatographic purification 193 of adenosine deaminase Affinity chromatographic purification 194 of histidinol dehydrogenase Purification of dehydrogenase 195... 

Example of a Protein Purification Scheme Purification of the Enzyme Xanthine Dehydrogenase from a Eungus... 

The specific activity is die total activity of die fracdon divided by the total protein in die fracdon. This value gives an indication of die increase in purity attained during die course of the purification as die samples become enriched for xandiine dehydrogenase protein. 

Most purification procedures for a particular protein are developed in an empirical manner, the overriding principle being purification of the protein to a homogeneous state with acceptable yield. Table 5.5 presents a summary of a purification scheme for a selected protein. Note that the specific activity of the protein (the enzyme xanthine dehydrogenase) in the immuno-affinity purified fraction (fraction 5) has been increased 152/0.108, or 1407 times the specific activity in the crude extract (fraction 1). Thus, xanthine dehydrogenase in fraction 5 versus fraction 1 is enriched more than 1400-fold by the purification procedure. 

The fact that adenosine and its derivatives are azo coupling components is used for immobilizing nicotinamide-adenine nucleotide (NAD+) for affinity chromatography purposes. In 12.58 NAD+ is bonded to a matrix through an azo bond. Compound 12.58 is used for the purification of dehydrogenase enzymes (Hocking and Harris, 1973). 

Uchicda, Y., Izai, K., Orii, T., Hashimoto, T. (1992). Novel fatty acid p-oxidation enzymes in rat liver mitochondria. II. Purification and properties of enoyl-coenzyme A (CoA) hydratase/3-hy-droxyacyl-CoA dehydrogenase/3-ketoacyl-CoA thiolase trifunctional protein. J. Biol. Chem. 267, 1034-1041. 

Purwantini E, L Daniels (1996) Purification of a novel coenzyme F42o-dependent glucose-6-phosphate dehydrogenase from Mycobacterium smegmatis. J Bacteriol 178 2861-2866. 

Drake HL. S-1 Hu, HG Wood (1980) Purification of carbon monoxide dehydrogenase, a nickel enzyme from Clostridium thermoaceticum. J Biol Chem 255 7174-7180. 

Yamamoto I, T Saiki, S-M Liu, LG Ljungdahl (1983) Purification and properties of NADP-dependent formate dehydrogenase from Clostridium thermoaceticum, a tungsten-selenium-iron protein. J Biol Chem 258 1826-1832. 

Bamforth CW, PJ Large (1977) Solubilization, partial purification and properties of A-methylglutamate dehydrogenase from Pseudomonas aminovorans. Biochem J 161 357-370. 

Hartel U, E Eckel, J Koch, G Fuchs, D, Linder, W Buckel (1993) Purification of glutaryl-CoA dehydrogenase from Pseudomonas sp., an enzyme involved in the anaerobic degradation of benzoate. Arch Microbiol 159 174-181. 

Hensgens CMH, J Vonck, J Van Beeumen, EFJ van Bruggen, TA Hansen (1993) Purification and characterization of an oxygen-labile, NAD-dependent alcohol dehydrogenase from Desulfovibrio gigas. J Bacterial 175 2859-2863. 

Tokimatsu T, Y Nagai, T Hattori, M Shimada (1998) Purification and characteristics of a novel cytochrome c dependent glyoxylate dehydrogenase from a wood-rotting fungus Tyromyces palustris. EEBS Lett 437 117-121. 

Axcell BC, PJ Geary (1973) The metabolism of benzene by bacteria. Purification and some properties of the ezyme cw-l,2-dihydroxycyclohexa-3,5-diene (nicotinamide adenine dinucleotide) oxidoreductase (ciT-benzene glycol dehydrogenase). Biochem J 136 927-934. 

Biegert T, U Altenschmidt, C Eckerskorn, G Euchs (1995) Purification and properties of benzyl alcohol dehydrogenase from a denitrifying Thauera sp. Arch Microbiol 163 418-423. 

Rogers JE, DT Gibson (1977) Purification and properties of cw-toluene dihydrodiol dehydrogenase from Pseudomonas putida. J Bacterial 130 1117-1124. 

Patel TR, DT Gibson (1974) Purification and properties of (+)-cZ5-naphthalene dihydrodiol dehydrogenase of Pseudomonas putida. Bacteriol 119 879-888. 

Lu Z-H, R Zhang, RB Diasio (1992) Purification and characterization of dihydropyrimidine dehydrogenase from human liver. J Biol Chem 267 17102-17109A. 

Nagel M, JR Andreesen (1990) Purification and characterization of the molybdoenzymes nicotinate dehydrogenase and 6-hydroxynicotinate dehydrogenase from Bacillus niacini. Arch Microbiol 154 605-613. 

Wagner R, R Cammack, JR Andreesen (1984) Purification and characterization of xanthine dehydrogenase from Clostridium acidiurici grown in the presence of selenium. Biochim Biophys Acta 791 63-74. 

Krupenko, S. A. and Wagner, G., Overexpression of functional hydrolase domain of rat liver 10-formyltetrahydrofolate dehydrogenase in Escherichia coli, Prot. Purif. Expression, 14, 146, 1998. 

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