scholarly journals The aromatic alcohol dehydrogenases in Pseudomonas putida N.C.I.B. 9869 grown on 3,5-xylenol and p-cresol

1978 ◽  
Vol 175 (2) ◽  
pp. 659-667 ◽  
Author(s):  
M J Keat ◽  
D J Hopper
Keyword(s):  
Alcohol Dehydrogenase ◽  
Pseudomonas Putida ◽  
The Other ◽  
Alcohol Dehydrogenases ◽  
Whole Cells ◽  
Aromatic Alcohol ◽  
Aromatic Alcohols ◽  
Hydroxybenzyl Alcohol

Whole cells of Pseudomonas putida N.C.I.B 9869, when grown on either 3,5-xylenol or p-cresol, oxidized both m- and p-hydroxybenzyl alcohols. Two distinct NAD+-dependent m-hydroxybenzyl alcohol dehydrogenases were purified from cells grown on 3,5-xylenol. Each is active with a range of aromatic alcohols, including both m- and p-hydroxybenzyl alcohol, but differ in their relative rates with the various substrates. An NAD+-dependent alcohol dehydrogenase was also partially purified from p-cresol grown cells. This too was active with m- and p-hydroxybenzyl alcohol and other aromatic alcohols, but was not identical with either of the other two dehydrogenases. All three enzymes were unstable, but were stabilized by dithiothreitol and all were inhibited with p-chloromercuribenzoate. All were specific for NAD+ and each was shown to catalyse conversion of alcohol into aldehyde.

scholarly journals Variation in the Direction of Selection Applied by Pentenol to the Alcohol Dehydrogenase Locus in Drosophila melanogaster

1977 ◽  
Vol 30 (3) ◽  
pp. 259 ◽  
Author(s):  
JG Oakeshott
Keyword(s):  
Drosophila Melanogaster ◽  
Alcohol Dehydrogenase ◽  
Null Allele ◽  
Developmental Time ◽  
The Other ◽  
Laboratory Population ◽  
Alcohol Dehydrogenases ◽  
Alcohol Dehydrogenase Activity ◽  
Active Alcohol ◽  
Selective Effects

This paper describes selective effects of pentenol-impregnated media on six genotypes at the alcohol dehydrogenase (Adh) locus in D. melanogaster. In the laboratory population studied, developmental times of pre-adults homozygous for an alcohol dehydrogenase 'null' allele increased with increasing pentenol concentrations. The developmental times of the other five genotypes, which produced active alcohol dehydrogenases, increased slightly at pentenol concentrations up to 0�0033%, but above this concentration they decreased markedly. In fact on 0�067% pentenol, the highest concentration tested, developmental times of these five genotypes were between 9 and 24 h less than their developmental times on media lacking penteno!. The magnitude of the reduction in developmental time differed significantly between genotypes and was positively correlated with alcohol dehydrogenase activity.

scholarly journals The purification and properties of p-cresol-(acceptor) oxidoreductase (hydroxylating), a flavocytochrome from Pseudomonas putida

1977 ◽  
Vol 167 (1) ◽  
pp. 155-162 ◽  
Author(s):  
D J Hopper ◽  
D G Taylor
Keyword(s):  
Pseudomonas Putida ◽  
Particulate Material ◽  
The Other ◽  
Methyl Group ◽  
Purification And Properties ◽  
Weight One ◽  
Dehydrogenation Reactions ◽  
Phenazine Methosulphate ◽  
Hydroxybenzyl Alcohol

The enzyme that catalyses the hydroxylation of the methyl group of p-cresol was purified from Pseudomonas putida. It has mol.wt. 115000 and appears to contain two subunits of equal molecular weight. One subunit is a c-type cytochrome and the other is a flavoprotein. Reduction of the cytochrome occurred on addition of substrate. The same enzyme catalyses both p-cresol hydroxylation and the further oxidation of the product, 4-hydroxybenzyl alcohol. The stoicheiometry of acceptor reduced per molecule of substrate oxidized is that for two dehydrogenation reactions. The Km for p-cresol is 7.3 × 10(-6) M and that for 4-hydroxybenzyl alcohol is 47.6 × 10(-6) M. The enzyme, which is assayed with phenazine methosulphate as electron acceptor, was stimulated by particulate material, which probably contains the acceptor in vivo.

scholarly journals Quinoprotein alcohol dehydrogenase from ethanol-grown Pseudomonas aeruginosa

1984 ◽  
Vol 223 (3) ◽  
pp. 921-924 ◽  
Author(s):  
B Groen ◽  
J Frank ◽  
J A Duine
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Amino Acid ◽  
Alcohol Dehydrogenase ◽  
Substrate Specificity ◽  
Aromatic Amino Acid ◽  
Pyrroloquinoline Quinone ◽  
The Other ◽  
Enzyme Molecule ◽  
Alcohol Dehydrogenases ◽  
Suicide Substrate

Cell-free extracts of Pseudomonas aeruginosa strains, grown on ethanol, showed dye-linked alcohol dehydrogenase activities. The enzyme responsible for this activity was purified to homogeneity. It appeared to contain two molecules of pyrroloquinoline quinone per enzyme molecule. In many respects, it resembled other quinoprotein alcohol dehydrogenases (EC 1.1.99.8), having a substrate specificity intermediate between that of methanol dehydrogenases and ethanol dehydrogenases in this group. On the other hand, it also showed dissimilarities: the enzyme was found to be a monomer (Mr 101 000), to need only one molecule of the suicide substrate cyclopropanol to become fully inactivated, and to have a different aromatic amino acid composition.

scholarly journals Characterization of a Pseudomonas putidaAllylic Alcohol Dehydrogenase Induced by Growth on 2-Methyl-3-Buten-2-ol

1999 ◽  
Vol 65 (6) ◽  
pp. 2622-2630 ◽  
Author(s):  
Vincent F. Malone ◽  
Amy J. Chastain ◽  
John T. Ohlsson ◽  
Loelle S. Poneleit ◽  
Michele Nemecek-Marshall ◽  
...  
Keyword(s):  
Alcohol Dehydrogenase ◽  
Benzyl Alcohol ◽  
Acinetobacter Calcoaceticus ◽  
Reduction Reaction ◽  
Terminal Sequence ◽  
Alcohol Dehydrogenases ◽  
Optimum Ph ◽  
Aromatic Alcohols ◽  
A Subunit

ABSTRACT We have been working to develop an enzymatic assay for the alcohol 2-methyl-3-buten-2-ol (232-MB), which is produced and emitted by certain pines. To this end we have isolated the soil bacteriumPseudomonas putida MB-1, which uses 232-MB as a sole carbon source. Strain MB-1 contains inducible 3-methyl-2-buten-1-ol (321-MB) and 3-methyl-2-buten-1-al dehydrogenases, suggesting that 232-MB is metabolized by isomerization to 321-MB followed by oxidation. 321-MB dehydrogenase was purified to near-homogeneity and found to be a tetramer (151 kDa) with a subunit mass of 37,700 Da. It catalyzes NAD+-dependent, reversible oxidation of 321-MB to 3-methyl-2-buten-1-al. The optimum pH for the oxidation reaction was 10.0, while that for the reduction reaction was 5.4. 321-MB dehydrogenase oxidized a wide variety of aliphatic and aromatic alcohols but exhibited the highest catalytic specificity with allylic or benzylic substrates, including 321-MB, 3-chloro-2-buten-1-ol, and 3-aminobenzyl alcohol. The N-terminal sequence of the enzyme contained a region of 64% identity with the TOL plasmid-encoded benzyl alcohol dehydrogenase of P. putida. The latter enzyme and the chromosomally encoded benzyl alcohol dehydrogenase ofAcinetobacter calcoaceticus were also found to catalyze 321-MB oxidation. These findings suggest that 321-MB dehydrogenase and other bacterial benzyl alcohol dehydrogenases are broad-specificity allylic and benzylic alcohol dehydrogenases that, in conjunction with a 232-MB isomerase, might be useful in an enzyme-linked assay for 232-MB.

Biodegradation of chlorophenols by immobilized pure-culture microorganisms

1996 ◽  
Vol 34 (10) ◽  
pp. 67-72 ◽  
Author(s):  
Lu Chih-Jen ◽  
Lee Chi-Mei ◽  
Huang Chiou-Zong
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Pseudomonas Putida ◽  
Pure Culture ◽  
The Other ◽  
Batch Reactors ◽  
Agrobacterium Radiobacter ◽  
Culture Cells ◽  
Lag Period

The biodegradation of phenol and chlorophenols by immobilized pure-culture cells was conducted by a series of batch reactors. The microorganisms used in this study were Pseudomonas putida, Psuedomonas testosteroni, Pseudomonas aeruginosa, and Agrobacterium radiobacter. All four species showed the ortho-cleavage pathway to metabolize chlorophenols. Among the four species, P. testosteroni, P. putida, and P. aeruginosa could effectively remove phenol at 200 mg/l. P. testosteroni could effectively remove 2-chlorophenol at 10mg/l. However, the other three species, P. putida, P. aeruginosa, and A. radiobacter, could not effectively remove 2-chlorophenol. Although 3-chlorophenol is a recalcitrant compound, P. testosteroni also could rapidly metabolize 3-chlorophenol at 10 mg/l. The removal of 4-chlorophenol at 10 mg/l by P. testosteroni reached 98% within one day. P. aeruginosa and A. radiobacter also could metabolize 4-chlorophenol after 2 and 7 days of lag period, respectively.

scholarly journals An ADH toolbox for raspberry ketone production from natural resources via a biocatalytic cascade

2021 ◽  
Author(s):  
Aileen Becker ◽  
Dominique Böttcher ◽  
Werner Katzer ◽  
Karsten Siems ◽  
Lutz Müller-Kuhrt ◽  
...  
Keyword(s):  
Alcohol Dehydrogenase ◽  
Genetically Modified Organisms ◽  
Optical Purity ◽  
High Yield ◽  
Alcohol Dehydrogenases ◽  
Flavor Compound ◽  
Raspberry Ketone ◽  
Cofactor Recycling ◽  
Cosmetic Industry ◽  
Key Points

Abstract Raspberry ketone is a widely used flavor compound in food and cosmetic industry. Several processes for its biocatalytic production have already been described, but either with the use of genetically modified organisms (GMOs) or incomplete conversion of the variety of precursors that are available in nature. Such natural precursors are rhododendrol glycosides with different proportions of (R)- and (S)-rhododendrol depending on the origin. After hydrolysis of these rhododendrol glycosides, the formed rhododendrol enantiomers have to be oxidized to obtain the final product raspberry ketone. To be able to achieve a high conversion with different starting material, we assembled an alcohol dehydrogenase toolbox that can be accessed depending on the optical purity of the intermediate rhododendrol. This is demonstrated by converting racemic rhododendrol using a combination of (R)- and (S)-selective alcohol dehydrogenases together with a universal cofactor recycling system. Furthermore, we conducted a biocatalytic cascade reaction starting from naturally derived rhododendrol glycosides by the use of a glucosidase and an alcohol dehydrogenase to produce raspberry ketone in high yield. Key points • LB-ADH, LK-ADH and LS-ADH oxidize (R)-rhododendrol • RR-ADH and ADH1E oxidize (S)-rhododendrol • Raspberry ketone production via glucosidase and alcohol dehydrogenases from a toolbox Graphical abstract

Are membrane properties essential for the circadian rhythm of Gonyaulax?

1984 ◽  
Vol 247 (2) ◽  
pp. R250-R256
Author(s):  
H. G. Scholubbers ◽  
W. Taylor ◽  
L. Rensing
Keyword(s):  
Circadian Rhythm ◽  
Phase Shift ◽  
Fluorescence Polarization ◽  
The Other ◽  
Membrane Properties ◽  
Response Curves ◽  
Whole Cells ◽  
Different Temperatures ◽  
Free Running ◽  
Free Running Period

Membrane properties of whole cells of Gonyaulax polyedra were measured by fluorescence polarization. Circadian changes of fluorescence polarization exist in exponentially growing cultures. They show an amplitude larger than that of stationary cultures, indicating that a part of the change is due to or amplified by an ongoing cell cycle. Measurements of parameters of the circadian glow rhythm were analyzed for possible correlation with the membrane data. Considerable differences (Q10 = 2.5-3.0) in fluorescence polarization were found in cultures kept at different temperatures ranging from 15 to 27.5 degrees C. The free-running period length at different temperatures, on the other hand, differed only slightly (Q10 = 0.9-1.1). Stationary cultures showed higher fluorescence polarization compared with growing cultures, whereas the free-running period lengths did not differ in cultures of various densities and growth rates. Temperature steps of different sign changed the fluorescence polarization slightly in different directions. The phase shift of 4-h pulses (-5, -9, +7 degrees C) resulted in maximal phase advances of 4, 6, and 2 h, respectively. The phasing of the phase-response curves was identical in all these experiments, a finding not to be expected if the pulses act via the measured membrane properties. Pulses of drugs that change the fluorescence polarization (e.g., chlorpromazine and lidocaine) did not or only slightly phase-shift the circadian rhythm.

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