1. Methylglyoxal synthase was purified over 1500-fold from glycerol-grown Escherichia coli K 12 strain CA 244. The enzyme inactivated by heat or proteolysis, had a molecular weight of approx. 67000, pH optimum 7.5 and specific for dihydroxyacetone phosphate with K(m) 0.47mm. 2. possibility that Schiff-base intermediate involved in the reaction mechanism investigated but not confirmed. 3. lost activity, especially at low temperature, could be stabilized P(i). Two binding sites P(i) may...

1. Intact cells of a non-fluorescent Pseudomonas grown with m-cresol, 2,5-xylenol, 3,5-xylenol, 3-ethyl-5-methylphenol or 2,3,5-trimethylphenol rapidly oxidized all these phenols to completion. 3-Hydroxybenzoate and 2,5-dihydroxybenzoate (gentisate) were also readily oxidized. 2. 3-Hydroxybenzoic acid 2,5-dihydroxybenzoic isolated as products m-cresol oxidation by inhibited alphaalpha'-bipyridyl. Alkyl-substituted 3-hydroxybenzoic acids alkyl-substituted gentisic formed similarly from...

A comparison of the oxidation rates various compounds by whole cells Pseudomonas putida 3, 5 indicated that m-cresol is metabolized to 3-hydroxybenzoate followed hydroxylation gentisate, ring-fission substrate, when grown with 5-xylenol. However, was growth similar experiments suggested a different pathway involving methyl-substituted catechol, and meta cleavage. Assays enzymes in cell-free extracts confirmed pathways are induced two substrates. 5-Xylenol-grown contained high levels...

p-Cresol methylhydroxylase from Pseudomonas putida, an anaerobic dehydrogenase that catalyses the oxidation of p-cresol to p-hydroxybenzyl alcohol and then p-hydroxybenzaldehyde, is enzyme great interest in several respects. One these fact its flavoprotein cytochrome c subunits may be reversibly dissociated with ease, full regeneration activity native properties on recombining components. Bisubstrate kinetic analysis unresolved gives parallel-line kinetics double-reciprocal plots, whereas...

Research Article| December 01 1968 Enzymic formation of D-malate D J Hopper; Hopper 1Department Biochemistry, University Minnesota, St Paul, Minn. 55101, U.S.A. Search for other works by this author on: This Site PubMed Google Scholar P Chapman; Chapman S Dagley Biochem (1968) 110 (4): 798–800. https://doi.org/10.1042/bj1100798 Views Icon Article contents Figures & tables Video Audio Supplementary Data Peer Review Share Facebook Twitter LinkedIn MailTo Cite Get Permissions Citation Hopper,...

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

In the hydroxylation of methyl group p-cresol by an enzyme from Pseudomonas putida oxygen atom is derived water. Although a second reaction same converts product, p-hydroxybenzyl alcohol, into aldehyde, alcohol enzyme-free intermediate.

Pseudomonas putida N.C.I.B. 9869, when grown on 3,5-xylenol, hydroxylates the methyl groups 3,5-xylenol and p-cresol by two different enzymes. 3,5-Xylenol methylhydroxylase, studied only in relatively crude extracts, requires NADH, is not active with inhibited cyanide, but CO. The methylhydroxylase an electron acceptor will act under anaerobic conditions. It was purified a flavocytochrome c of mol.wt. approx. 114,000 consisting subunits equal size. enzyme catalyses hydroxylation (Km 16...

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXT8.alpha.-(O-Tyrosyl)flavin adenine dinucleotide, the prosthetic group of bacterial p-cresol methylhydroxylaseWilliam McIntire, Dale E. Edmondson, David J. Hopper, and Thomas P. SingerCite this: Biochemistry 1981, 20, 11, 3068–3075Publication Date (Print):May 1, 1981Publication History Published online1 May 2002Published inissue 1 1981https://pubs.acs.org/doi/10.1021/bi00514a013https://doi.org/10.1021/bi00514a013research-articleACS...

1. Cell-free extracts, prepared from a non-fluorescent Pseudomonas grown on m-cresol, oxidized gentisate and certain alkyl-substituted gentisates with the consumption of 1 mol oxygen formation pyruvate substrate. 2. In addition to pyruvate, malate was formed gentisate; citramalate 3-methylgentisate 4-methylgentisate; 2,3-dimethylmalate 3,4-dimethylgentisate. 3. One enantiomer, d-(-)-citramalate, enzymically 3-methylgentisate, 4-methylgentisate citraconate. l-(+)-Citramalate mesaconate by...

1. Measurements of the rates oxidation various compounds by a fluorescent Pseudomonas indicated that metabolism 2,4-xylenol was initiated methyl group para to hydroxyl group. 2. 4-Hydroxy-3-methylbenzoic acid isolated as product cells inhibited with alphaalpha'-bipyridyl. 3. 4-Hydroxyisophthalic accumulated at low oxygen concentrations when either or 4-hydroxy-3-methylbenzoic oxidized grown 2,4-xylenol. 4. When supplemented NADH, but not NADPH, cell extracts readily....

The fungus Aspergillus fumigatus ATCC 28282 was shown to grow on p -cresol as its sole source of carbon and energy. A pathway for metabolism this compound proposed. This has protocatechuate the ring-fission substrate with cleavage by an ortho -fission pathway. formed two alternative routes, either initial attack methyl group, which is oxidized carboxyl, followed ring-hydroxylation, or ring-hydroxylation first step subsequent oxidation 4-methylcatechol acid. elucidated from several pieces...

The enzyme 4-ethylphenol methylenehydroxylase was purified from Pseudomonas putida JD1 grown on 4-ethylphenol. It is a flavocytochrome c for which the Mr found to be 120,000 by ultracentrifuging and 126,000 gel filtration. consists of two flavoprotein subunits each 50,000 cytochrome 10,000. redox potential 240 mV. Hydroxylation proceeds dehydrogenation hydration give 1-(4'-hydroxyphenyl)ethanol, also dehydrogenated same 4-hydroxyacetophenone. will hydroxylate p-cresol but more active with...

Constitutive synthesis of enzymes responsible for methyl group oxidation in 3,5-xylenol degradation and an associated p-cresol methylhydroxylase Pseudomonas putida NCIB 9869 was shown by their retention at high specific activities cells transferred from medium to glutamate medium. The other the pathway declined upon removal aromatic substrate, consistent with inducible control. Specific methyl-oxidizing showed eventual decline concomitant a decrease fraction bacteria capable growth...

Three independent mechanisms are described that contaminate the phase-modulated pump beam of an optical frequency reference stabilized by modulation transfer spectroscopy (MTS) with residual amplitude (RAM). The electro-optic modulator, geometry and absorption saturated medium all separately generate undesired RAM degrades accuracy reference. An analysis is presented shows how shifts introduced different can be evaluated in typical MTS set-ups minimized. also detector phase used to measure...

A bacterium, strain PC-07, previously isolated as part of a coculture capable growing on p-cresol under anaerobic conditions with nitrate the acceptor was identified an Achromobacter sp. The first enzyme pathway, methylhydroxylase, which converts its substrate into p-hydroxybenzyl alcohol, purified. had Mr 130,000 and spectrum flavocytochrome. It composed flavoprotein subunits 54,000 cytochrome 12,500. midpoint redox potential 232 mV. Km kcat for were 21 microM 112 s-1 respectively,...

SUMMARY: A bacterium capable of growth on 4-ethylphenol was isolated from soil and identified as Pseudomonas putida. Intact cells grown rapidly oxidized 4-hydroxyaceto-phenone well substrate the also 4-hydroxyacetophenone. The initial enzymes for catabolism were still present, although at lower activities, in succinate-grown which to Extracts 4-ethylphenol-grown 4-hydroxyacetophenone when provided with NADPH. When this activity partially purified a stoichiometry 1 μmol O2 consumed per...

Enzymic hydroxylation of 4-ethylphenol by (a) Pseudomonas putida and (b) highly purified p-cresol methylhydroxylase gave optically active 1-(4′-hydroxyphenyl)-ethanol. The products were transformed into the phenolic methyl ethers shown to contain 69.5% 65.6%, respectively, (S)-(-)-isomer. stereochemistry reaction is discussed in terms three distinct steps occurring at site enzyme.

Extracts of a fluorescent species Pseudomonas grown with m -cresol, degrade gentisic acid without isomerization the ring-fission compound, maleylpyruvate, to give eventually d -malate and pyruvate. -Malate is also growth substrate. l but not oxidized by particulate enzyme requiring nicotinamide adenine dinucleotide (NAD) or phosphate (NADP). NAD- NADP-linked malate dehydrogenases are absent cells contain an NADP-dependent -malic enzyme. Exposure exogenous induces NAD-dependent enzyme,...

The electrochemistry of the enzyme, sulfide:cytochrome c oxidoreductase, also known as flavocytochrome c552 from purple sulfur bacterium, Chromatium vinosum, has been studied using several modified electrodes. Direct electron transfer between heme and an electrode is observed in presence a redox-inactive cationic species which promotes voltammetry enzyme. Quasi-reversible was achieved aminoglycoside, neomycin, promoter at either gold or polished edge-plane graphite electrode. Further...