scholarly journals Quinohaemoprotein alcohol dehydrogenase apoenzyme from Pseudomonas testosteroni

1986 ◽  
Vol 234 (3) ◽  
pp. 611-615 ◽  
Author(s):  
B W Groen ◽  
M A G van Kleef ◽  
J A Duine
Keyword(s):  
Absorption Spectrum ◽  
Cytochrome C ◽  
Alcohol Dehydrogenase ◽  
Secondary Alcohols ◽  
Primary Alcohols ◽  
Optimal Activity ◽  
Alcohol Dehydrogenase Activity ◽  
Artificial Electron Acceptor ◽  
The Common ◽  
Pseudomonas Testosteroni

Cell-free extracts of Pseudomonas testosteroni, grown on alcohols, contain quinoprotein alcohol dehydrogenase apoenzyme, as was demonstrated by the detection of dye-linked alcohol dehydrogenase activity after the addition of PQQ (pyrroloquinoline quinone). The apoenzyme was purified to homogeneity, and the holoenzyme was characterized. Primary alcohols (except methanol), secondary alcohols and aldehydes were substrates, and a broad range of dyes functioned as artificial electron acceptor. Optimal activity was observed at pH 7.7, and the presence of Ca2+ in the assay appeared to be essential for activity. The apoenzyme was found to be a monomer (Mr 67,000 +/- 5000), with an absorption spectrum similar to that of oxidized cytochrome c. After reconstitution to the holoenzyme by the addition of PQQ, addition of substrate changed the absorption spectrum to that of reduced cytochrome c, indicating that the haem c group participated in the enzymic mechanism. The enzyme contained one haem c group, and full reconstitution was achieved with 1 mol of PQQ/mol. In view of the aberrant properties, it is proposed to distinguish the enzyme from the common quinoprotein alcohol dehydrogenases by using the name ‘quinohaemoprotein alcohol dehydrogenase’. Incorporation of PQQ into the growth medium resulted in a significant shortening of lag time and increase in growth rate. Therefore PQQ appears to be a vitamin for this organism during growth on alcohols, reconstituting the apoenzyme to a functional holoenzyme.

Oxygen toxicity and carbon deprivation in Astasia longa

1970 ◽  
Vol 48 (3) ◽  
pp. 251-258 ◽  
Author(s):  
Nicole Bégin-Heick
Keyword(s):  
Cytochrome C ◽  
Alcohol Dehydrogenase ◽  
Dehydrogenase Activity ◽  
Cellular Respiration ◽  
Endogenous Respiration ◽  
Oxidoreductase Activity ◽  
Oxygen Treatment ◽  
Astasia Longa ◽  
Alcohol Dehydrogenase Activity ◽  
Nadh Cytochrome

The changes in metabolic activity of Astasia longa during oxygen treatment and carbon deprivation were examined. In many respects (growth, cell composition, and some enzymatic activities) the two treatments produce similar effects. Other parameters such as cellular respiration are affected differently. Oxygen treatment inhibits the ethanol-stimulated respiration but affects endogenous respiration very little. At the same time, the total alcohol dehydrogenase activity of the cell is increased at least 10-fold. In starved cells, the endogenous respiration is decreased parallel to the cellular content of carbohydrate reserves, but the ability of the cell to utilize ethanol stays constant. The NADH : cytochrome c oxidoreductase activity of the oxygen-treated cells is decreased by 70% whereas this activity is decreased less than 30% in starved cells. It is proposed that the inhibition of the NADH : cytochrome c oxidoreductase activity is the primary activity affected by oxygen treatment in both acetate- and ethanol-grown cells. The characteristic effects of oxygen treatment on ethanol-grown cells are probably the result of the combination of increased NAD-dependent alcohol dehydrogenase activity and decreased NADH : cytochrome c oxidoreductase activity. Such a modification of these two activities would tend to keep NAD in the reduced state and therefore limit cellular reactions which require oxidized NAD.

scholarly journals Novel NADP-linked alcohol–aldehyde/ketone oxidoreductase in thermophilic ethanologenic bacteria

1981 ◽  
Vol 195 (1) ◽  
pp. 183-190 ◽  
Author(s):  
R J Lamed ◽  
J G Zeikus
Keyword(s):  
Metal Ion ◽  
Polyacrylamide Gel Electrophoresis ◽  
Secondary Alcohol ◽  
Sodium Dodecyl ◽  
Secondary Alcohols ◽  
Metal Ion Binding ◽  
Primary Alcohols ◽  
Cell Extracts ◽  
Alcohol Dehydrogenase Activity ◽  
Clostridium Thermohydrosulfuricum

An NADP-specific alcohol--aldehyde/ketone oxidoreductase was detected in cell extracts of Thermoanaerobium brockii and Clostridium thermohydrosulfuricum, but not in Thermobacteroides acetoethylicus or Clostridium thermocellum. The enzyme was purified from Ta. brockii by differential procedures that included heat treatment and an affinity-chromatography step on Blue Dextran--Sepharose. The 44-fold-purified enzyme displayed one band (mol.wt. approx. 40000) after sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. The enzyme had a broad substrate specificity that included linear and branched primary alcohols, linear and cyclic secondary alcohols, linear and cyclic ketones, and acetaldehyde. The NADP-specific alcohol--aldehyde/ketone oxidoreductase was considerably more active towards secondary alcohols than towards other substrates. The enzyme had remarkable stability to heating at 86 degrees C for 70 min, but was rapidly denatured on boiling. Secondary-alcohol dehydrogenase activity displayed a noticeable inflexion point at 50 degrees C in Arrhenius plots and a high Q10 value (greater than 2.0). The enzyme was inactivated by the thiol-blocking reagent p-chloromercuribenzoate, but was not significantly inhibited by common metal-ion-binding agents. The NADP-linked alcohol--aldehyde/ketone oxidoreductase of Ta. brockii appears to have properties distinct from those of previously described primary- and secondary-alcohol dehydrogenases.

scholarly journals Biochemical properties of alcohol dehydrogenase from Drosophila lebanonensis

1986 ◽  
Vol 235 (2) ◽  
pp. 481-490 ◽  
Author(s):  
J O Winberg ◽  
R Hovik ◽  
J S McKinley-McKee ◽  
E Juan ◽  
R Gonzalez-Duarte
Keyword(s):  
Alcohol Dehydrogenase ◽  
Active Site ◽  
Active Sites ◽  
Electrophoretic Pattern ◽  
Maximum Velocity ◽  
Cathode Side ◽  
Secondary Alcohols ◽  
Starch Gel Electrophoresis ◽  
Primary Alcohols ◽  
Rate Limiting

Purified Drosophila lebanonensis alcohol dehydrogenase (Adh) revealed one enzymically active zone in starch gel electrophoresis at pH 8.5. This zone was located on the cathode side of the origin. Incubation of D. lebanonensis Adh with NAD+ and acetone altered the electrophoretic pattern to more anodal migrating zones. D. lebanonensis Adh has an Mr of 56,000, a subunit of Mr of 28 000 and is a dimer with two active sites per enzyme molecule. This agrees with a polypeptide chain of 247 residues. Metal analysis by plasma emission spectroscopy indicated that this insect alcohol dehydrogenase is not a metalloenzyme. In studies of the substrate specificity and stereospecificity, D. lebanonensis Adh was more active with secondary than with primary alcohols. Both alkyl groups in the secondary alcohols interacted hydrophobically with the alcohol binding region of the active site. The catalytic centre activity for propan-2-ol was 7.4 s-1 and the maximum velocity of most secondary alcohols was approximately the same and indicative of rate-limiting enzyme-coenzyme dissociation. For primary alcohols the maximum velocity varied and was much lower than for secondary alcohols. The catalytic centre activity for ethanol was 2.4 s-1. With [2H6]ethanol a primary kinetic 2H isotope effect of 2.8 indicated that the interconversion of the ternary complexes was rate-limiting. Pyrazole was an ethanol-competitive inhibitor of the enzyme. The difference spectra of the enzyme-NAD+-pyrazole complex gave an absorption peak at 305 nm with epsilon 305 14.5 × 10(3) M-1 × cm-1. Concentrations and amounts of active enzyme can thus be determined. A kinetic rate assay to determine the concentration of enzyme active sites is also presented. This has been developed from active site concentrations established by titration at 305 nm of the enzyme and pyrazole with NAD+. In contrast with the amino acid composition, which indicated that D. lebanonensis Adh and the D. melanogaster alleloenzymes were not closely related, the enzymological studies showed that their active sites were similar although differing markedly from those of zinc alcohol dehydrogenases.

Observations on the histochemical distribution of alcohol dehydrogenase

1965 ◽  
Vol s3-106 (76) ◽  
pp. 289-297
Author(s):  
M. M. FERGUSON
Keyword(s):  
Alcohol Dehydrogenase ◽  
Secondary Alcohols ◽  
Primary Alcohols ◽  
Straight Chain ◽  
Tertiary Alcohols ◽  
Polyhydric Alcohols ◽  
Water Side ◽  
Monohydric Alcohols ◽  
Branched Chain ◽  
Ethereal Oxygen

Alcohol dehydrogenase capable of utilizing primary alcohols (straight and branched chain, unsaturated and cellosolves), secondary alcohols, polyhydric alcohols but not tertiary alcohols is described in the rat brain, lung, heart, liver, kidney, gut, spleen, pancreas, uterus, and seminal vesicle. With the straight chain primary alcohols the extent to which the alcohols were used increased relative to increasing chain length until the optimum length of six to seven carbon atoms had been reached, whereafter activity decreased, probably due to the insolubility of the higher members of the series in water. Side groups on the chain, double bonds and ethereal oxygen atoms, all had the effect of hindering the ease of dehydrogenation of the alcohol. Secondary alcohols were suitable as substrates; however, tertiary alcohols did not seem to be satisfactory for the demonstration of alcohol dehydrogenase. Polyhydric alcohols were suitable substrates although they were not as well used as were the monohydric alcohols. Furfuryl alcohol dehydrogenase was found in all tissues and is apparently a separate enzyme.

scholarly journals Physiological Significance of the Alcohol Dehydrogenase Polymorphism in Larvae of Drosophila

Genetics ◽  
1987 ◽  
Vol 117 (1) ◽  
pp. 75-84
Author(s):  
Pieter W H Heinstra ◽  
Willem Scharloo ◽  
George E W Thörig
Keyword(s):  
Alcohol Dehydrogenase ◽  
Rank Order ◽  
Adult Survival ◽  
Alcohol Metabolism ◽  
Oxidation Rates ◽  
Alcohol Dehydrogenase Activity ◽  
The Common ◽  
The Relationship ◽  
Ethanol Elimination

ABSTRACT This study deals with biochemical and metabolic-physiological aspects of the relationship between variation in in vivo alcohol dehydrogenase activity and fitness in larvae homozygous for the alleles Adh71k, AdhF, AdhS, of Drosophila melanogaster, and for the common Adh allele of Drosophila simulans. The Adh genotypes differ in the maximum oxidation rates of propan-2-ol into acetone in vivo. There are smaller differences between the Adh genotypes in rates of ethanol elimination. Rates of accumulation of ethanol in vivo are negatively associated with larval-to-adult survival of the Adh genotypes. The rank order of the maximum rates of the ADHs in elimination of propan-2-ol, as well as ethanol, is ADH-71k > ADH-F > ADH-S- > simulans-ADH. The ratio of this maximum rate to ADH quantity reveals the rank order of ADH-S > ADH-F > ADH-71k > simulans-ADH, suggesting a compensation for allozymic efficiency by the ADH quantity in D. melanogaster.—Our findings show that natural selection may act on the Adh polymorphism in larvae via differences in rates of alcohol metabolism.

Purification and properties of alcohol dehydrogenase from a mutant strain of Candida guilliermondii deficient in one form of the enzyme

1992 ◽  
Vol 38 (9) ◽  
pp. 953-957 ◽  
Author(s):  
Retno Indrati ◽  
Yoshiyuki Ohta
Keyword(s):  
Alcohol Dehydrogenase ◽  
Gel Filtration ◽  
Kinetic Studies ◽  
Candida Guilliermondii ◽  
Secondary Alcohols ◽  
Primary Alcohols ◽  
Subunit Molecular Weight ◽  
Purification And Properties ◽  
A Subunit ◽  
Better Than

Alcohol dehydrogenase (ADH1) was purified from Candida guilliermondii strain B10-05 to homogeneity, using affinity chromatography on triazine dyes and gel filtration. The enzyme was tetrameric, with a subunit molecular weight of 38 000. The purified enzyme oxidized primary and secondary alcohols, although it preferred primary alcohols. Its activity toward secondary alcohols was better than those of other yeast ADH; however, the enzyme was less sensitive toward inhibitors. Kinetic studies indicated that C. guilliermondii ADH1 oxidized ethanol by an ordered bi–bi mechanism, with NAD as the first substrate fixed. Key words: Candida guilliermondii, alcohol dehydrogenase, ADH1, tetrameric.

ALCOHOL DEHYDROGENASE ACTIVITY IN ISOLATED VACUOLES OF RED BEET ROOT CELLS

Tsitologiya ◽  
2018 ◽  
Vol 60 (6) ◽  
pp. 469-475
Author(s):  
O. D. Nimaeva ◽  
◽  
E. V. Pradedova ◽  
A. B. Karpova ◽  
R. K. Salyaev ◽  
...  
Keyword(s):  
Alcohol Dehydrogenase ◽  
Dehydrogenase Activity ◽  
Root Cells ◽  
Beet Root ◽  
Red Beet ◽  
Alcohol Dehydrogenase Activity

Revisiting Sodium Hypochlorite Pentahydrate (NaOCl·5H2O) for the Oxidation of Alcohols in Acetonitrile without Nitroxyl Radicals

Synlett ◽  
2018 ◽  
Vol 29 (18) ◽  
pp. 2404-2407 ◽  
Author(s):  
Tsunehisa Hirashita ◽  
Yuto Sugihara ◽  
Shota Ishikawa ◽  
Yohei Naito ◽  
Yuta Matsukawa ◽  
...  
Keyword(s):  
Sodium Hypochlorite ◽  
Nitroxyl Radicals ◽  
Secondary Alcohols ◽  
Primary Alcohols ◽  
Oxidation Of Alcohols

Sodium hypochlorite pentahydrate (NaOCl·5H2O) is capable of oxidizing alcohols in acetonitrile at 20 °C without the use of catalysts. The oxidation is selective to allylic, benzylic, and secondary alcohols. ­Aliphatic primary alcohols are not oxidized.

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