A Glycerol Dehydrogenase From Escherichia Coli

Asnis, Robert E.; Brodie, Arnold F.

doi:10.1016/s0021-9258(19)52625-4

Cited by 67 publications

(7 citation statements)

References 8 publications

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“…The properties found in common between the two enzymes were a pH optimum of about 10 and a high degree of sensitivity to Cu2". No substrate specificity was undertaken in the previous work, except that the enzyme was shown to catalyze the reduction of dihydroxyacetone with NADH, and therefore the attack was also at C2 (1). Unfortunately, strain ECFS seems no longer available.…”

Section: Discussionmentioning

confidence: 99%

Purification and properties of a nicotinamide adenine dinucleotide-linked dehydrogenase that serves an Escherichia coli mutant for glycerol catabolism

Tang¹,

Ruch²,

Lin³

1979

J Bacteriol

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Glycerol:NAD+2-OXIDOREDUCTASE (EC 1.1.1.6) was purified to homogeneity from a mutant of Escherichia coli K12 that uses this enzyme, instead of ATP:glycerol 3-phosphotransferase (EC 2.7.1.30), as the first enzyme for the dissimilation of glycerol. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate shows a subunit of 39,000 daltons. During electrophoresis under nondenaturing conditions, the protein migrates as two bands. These two forms, both of which are enzymatically active, appear to be dimers and octomers of the same subunit. The optimal pH for the oxidation of glycerol is about 10, and that for the reduction of dihydroxyacetone is about 6. Glycerol dehydrogenation is highly activated by NH4+, K+, or Rb+, but strongly inhibited by N-ethylmalemide, 8-hydroxyquinoline, 1,10-phenanthroline, Cu2+, and Ca2+. The enzyme exhibits a broad substrate specificity. In addition to glycerol, it act on 1,2-propanediol and several of its analogs.

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Section: Discussionmentioning

confidence: 99%

Purification and properties of a nicotinamide adenine dinucleotide-linked dehydrogenase that serves an Escherichia coli mutant for glycerol catabolism

Tang¹,

Ruch²,

Lin³

1979

J Bacteriol

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“…As an alternative to column chromatography, the recombinant EcoGroDHase was purified by heat-shock treatment. Briefly, the soluble fraction was heated for 10 min at 60 • C followed by incubation on an ice-cold bath [5]. After 30 min, precipitated proteins were separated by centrifuging 10 min at 4 • C and 30,000 × g. Active fractions were pooled, fractionated and stored at −80 • C until use.…”

Section: Enzyme Purificationmentioning

confidence: 99%

“…GroDHase activity was assayed as described previously [5], with minor modifications. In the direction of Gro oxidation, the standard medium contained 100 mM CAPS-NaOH pH 10.5, 5 mM NAD + , and 500 mM Gro.…”

Section: Activity Assaymentioning

confidence: 99%

“…EcoGroDHase is encoded by the gldA gene (1104 bp), is a polypeptide of ∼40 kDa, and in solution it forms homodimeric and homooctameric structures [4]. Nearly 60 years ago, Asnis and Brodie [5] purified this enzyme directly from the bacterium using a simple heat-shock protocol. In the last decade, the E. coli gldA gene was recombinantly expressed in order to improve either the capacity of redox cofactor regeneration in reverse micelles [6] or to reach a superior electrocatalytic performance of E. coli cells [7].…”

Section: Introductionmentioning

confidence: 99%

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Production and characterization of Escherichia coli glycerol dehydrogenase as a tool for glycerol recycling

Piattoni

Figueroa

Diez

et al. 2013

Process Biochemistry

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“…Glycerol dehydrogenase (EC 1.1.1.6) catalyzes the NAD+-linked1 oxidation of glycerol to dihydroxyacetone and provides microorganisms an effective, ATP-independent, metabolic pathway for glycerol utilization. This enzyme has been isolated from a variety of bacterial sources including Escherichia coli (Asnis & Brodie, 1953), Aerobacter aerogenes (Burton & Kaplan, 1953;Rush et al, 1957), Klebsiella aerogenes (Lin & Magasanik, 1960), Bacillus megaterium (Scharschmidt et al, 1984), and Bacillus stearothermophilus (Spencer et al, 1989). The enzyme from Cellulomonas sp.…”

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confidence: 99%

Isotopic Analysis of the Reaction Catalyzed by Glycerol Dehydrogenase

Leichus¹,

Blanchard²

1994

Biochemistry

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Glycerol dehydrogenase catalyzes the reversible NAD(+)-dependent oxidation of glycerol to form dihydroxyacetone. Initial velocity, product, and dead-end inhibition studies performed for the forward and reverse reactions support an ordered kinetic mechanism with NAD+ binding first and NADH released last. A monovalent cation is required for enzymatic activity and glycerol binding, with K+ having the highest activity as measured by V. The pH dependence of the kinetic parameters V and V/Kglycerol, as well as the temperature dependence of the V pH profile, suggested that an enzymic carboxylate group functions as a base in catalysis. The pH dependence of the primary deuterium kinetic isotope effect shows that DV/Kglycerol increases from a pH-independent value of 1.15 at high pH values to a pH-independent value of 2.44 at low pH values. DV exhibits a similar pH dependence, increasing from a pH-independent value of 2.57 at high pH values to a pH independent value of 4.88 at low pH values. A chemical mechanism for enzymatic glycerol oxidation is proposed based on the data.

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A Glycerol Dehydrogenase From Escherichia Coli

Cited by 67 publications

References 8 publications

Purification and properties of a nicotinamide adenine dinucleotide-linked dehydrogenase that serves an Escherichia coli mutant for glycerol catabolism

Purification and properties of a nicotinamide adenine dinucleotide-linked dehydrogenase that serves an Escherichia coli mutant for glycerol catabolism

Production and characterization of Escherichia coli glycerol dehydrogenase as a tool for glycerol recycling

Isotopic Analysis of the Reaction Catalyzed by Glycerol Dehydrogenase

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