Oxalate Oxidase from Barley Roots: Purification to Homogeneity and Study of Some Molecular, Catalytic, and Binding Properties

Kotsira, Vicky Ph.; Clonis, Yannis D.

doi:10.1006/abbi.1997.9896

Cited by 76 publications

(88 citation statements)

References 39 publications

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“…2D. The catalytic efficiency (V max /K m ) increases continuously to low pH, rather than reaching a maximum at pH 4, as previously described for turnover at 37°C using a peroxidase-coupled assay (25). The K m evaluated from initial velocity data exhibits a strong pH dependence ( Fig.…”

Section: Steady State Kineticssupporting

confidence: 62%

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Burst Kinetics and Redox Transformations of the Active Site Manganese Ion in Oxalate Oxidase

Whittaker

Pan

Yukl

et al. 2007

Journal of Biological Chemistry

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Oxalate oxidase (EC 1.2.3.4) catalyzes the oxidative cleavage of oxalate to carbon dioxide and hydrogen peroxide. In this study, unusual nonstoichiometric burst kinetics of the steady state reaction were observed and analyzed in detail, revealing that a reversible inactivation process occurs during turnover, associated with a slow isomerization of the substrate complex. We have investigated the underlying molecular mechanism of this kinetic behavior by preparing recombinant barley oxalate oxidase in three distinct oxidation states (Mn(II), Mn(III), and Mn(IV)) and producing a nonglycosylated variant for detailed biochemical and spectroscopic characterization. Surprisingly, the fully reduced Mn(II) form, which represents the majority of the as-isolated native enzyme, lacks oxalate oxidase activity, but the activity is restored by oxidation of the metal center to either Mn(III) or Mn(IV) forms. All three oxidation states appear to interconvert under turnover conditions, and the steady state activity of the enzyme is determined by a balance between activation and inactivation processes. In O 2 -saturated buffer, a turnover-based redox modification of the enzyme forms a novel superoxidized mononuclear Mn(IV) biological complex. An oxalate activation role for the catalytic metal ion is proposed based on these results.

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Section: Steady State Kineticssupporting

confidence: 62%

“…2B) rather than the initial velocity, which is the typical substrate inhibition pattern. An earlier study anecdotally described substrate inhibition at oxalate concentrations above 4 mM (25). However, in subsequent work, no substrate inhibition was detected on the initial velocity (V i ) (8).…”

Section: Discussionmentioning

confidence: 94%

Burst Kinetics and Redox Transformations of the Active Site Manganese Ion in Oxalate Oxidase

Whittaker

Pan

Yukl

et al. 2007

Journal of Biological Chemistry

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“…The molecular weight of the enzyme was 58,500 as determined by SDS-PAGE (Fig. 5), whereas others reported an oxalate oxidase of rootlets of barley that is a pentamer with subunit molecular weight of 25,000 13 . Further work is needed to explain this discrepancy.…”

Section: Partial Purification and Properties Of Oxalate Oxidasementioning

confidence: 92%

Oxalate and Oxalate Oxidase in Malt

Kanauchi

Milet

Bamforth

2009

Journal of the Institute of Brewing

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Barley kernels contain a single oxalate oxidase located in the embryo and aleurone. It is already present in substantial quantities in unmalted grain and increases in activity during germination. It displays a very broad pH optimum: the optimum was at pH 4.0, but the enzyme still displayed more than 50% of its activity at pH 7. Oxalate oxidase is highly resistant to heat. However, its low affinity for oxygen suggests that it probably does not play a major role in the consumption of oxygen in mashing. The decrease in oxalic acid levels late in germination may be a result of oxalate oxidase action. Oxalic acid was not detected in raw barley.

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“…Some of these kits (e.g., the Sigma kit) utilize an enzyme isolated from barley roots, and although they are quick and easy to use, there is a continuous effort to improve the accuracy and efficiency of the assay (175,191,228). Such efforts will benefit from recently obtained data regarding fundamental biochemical and structural analysis of the barley enzyme itself (161,162,238,310) and from the finding (173) that the extremely well characterized wheat germin is also an OXO. This discovery was the culmination of the second important research track, one which started in the early 1980s, during which the GF-2.8 germin (gi121129) was found to be an apoplastic, multimeric (310), glycosylated (135) enzyme with extreme resistance to heat and to chemical degradation by protease or hydrogen peroxide.…”

Section: Germin and Germin-like Proteins From Higher Plantsmentioning

confidence: 99%

Microbial Relatives of the Seed Storage Proteins of Higher Plants: Conservation of Structure and Diversification of Function during Evolution of the Cupin Superfamily

Dunwell

Khuri

Gane

2000

Microbiol Mol Biol Rev

310

297

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SUMMARY This review summarizes the recent discovery of the cupin superfamily (from the Latin term “cupa,” a small barrel) of functionally diverse proteins that initially were limited to several higher plant proteins such as seed storage proteins, germin (an oxalate oxidase), germin-like proteins, and auxin-binding protein. Knowledge of the three-dimensional structure of two vicilins, seed proteins with a characteristic β-barrel core, led to the identification of a small number of conserved residues and thence to the discovery of several microbial proteins which share these key amino acids. In particular, there is a highly conserved pattern of two histidine-containing motifs with a varied intermotif spacing. This cupin signature is found as a central component of many microbial proteins including certain types of phosphomannose isomerase, polyketide synthase, epimerase, and dioxygenase. In addition, the signature has been identified within the N-terminal effector domain in a subgroup of bacterial AraC transcription factors. As well as these single-domain cupins, this survey has identified other classes of two-domain bicupins including bacterial gentisate 1,2-dioxygenases and 1-hydroxy-2-naphthoate dioxygenases, fungal oxalate decarboxylases, and legume sucrose-binding proteins. Cupin evolution is discussed from the perspective of the structure-function relationships, using data from the genomes of several prokaryotes, especially Bacillus subtilis. Many of these functions involve aspects of sugar metabolism and cell wall synthesis and are concerned with responses to abiotic stress such as heat, desiccation, or starvation. Particular emphasis is also given to the oxalate-degrading enzymes from microbes, their biological significance, and their value in a range of medical and other applications.

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Oxalate Oxidase from Barley Roots: Purification to Homogeneity and Study of Some Molecular, Catalytic, and Binding Properties

Cited by 76 publications

References 39 publications

Burst Kinetics and Redox Transformations of the Active Site Manganese Ion in Oxalate Oxidase

Burst Kinetics and Redox Transformations of the Active Site Manganese Ion in Oxalate Oxidase

Oxalate and Oxalate Oxidase in Malt

Microbial Relatives of the Seed Storage Proteins of Higher Plants: Conservation of Structure and Diversification of Function during Evolution of the Cupin Superfamily

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