Oxalate Decarboxylase and Oxalate Oxidase Activities Can Be Interchanged with a Specificity Switch of up to 282 000 by Mutating an Active Site Lid†,‡

Biochemistry ◽  
2007 ◽  
Vol 46 (43) ◽  
pp. 12327-12336 ◽  
Author(s):  
Matthew R. Burrell ◽  
Victoria J. Just ◽  
Laura Bowater ◽  
Shirley A. Fairhurst ◽  
Laura Requena ◽  
...  
Keyword(s):  
Active Site ◽  
Oxalate Oxidase ◽  
Oxalate Decarboxylase

scholarly journals The identity of the active site of oxalate decarboxylase and the importance of the stability of active-site lid conformations1

2007 ◽  
Vol 407 (3) ◽  
pp. 397-406 ◽  
Author(s):  
Victoria J. Just ◽  
Matthew R. Burrell ◽  
Laura Bowater ◽  
Iain McRobbie ◽  
Clare E. M. Stevenson ◽  
...  
Keyword(s):  
Crystal Structures ◽  
Active Site ◽  
Structural Information ◽  
Side Reaction ◽  
Oxalate Oxidase ◽  
Decarboxylase Activity ◽  
Oxalate Decarboxylase ◽  
Catalytically Active ◽  
The Stability ◽  
Kinetics Of

Oxalate decarboxylase (EC 4.1.1.2) catalyses the conversion of oxalate into carbon dioxide and formate. It requires manganese and, uniquely, dioxygen for catalysis. It forms a homohexamer and each subunit contains two similar, but distinct, manganese sites termed sites 1 and 2. There is kinetic evidence that only site 1 is catalytically active and that site 2 is purely structural. However, the kinetics of enzymes with mutations in site 2 are often ambiguous and all mutant kinetics have been interpreted without structural information. Nine new site-directed mutants have been generated and four mutant crystal structures have now been solved. Most mutants targeted (i) the flexibility (T165P), (ii) favoured conformation (S161A, S164A, D297A or H299A) or (iii) presence (Δ162–163 or Δ162–164) of a lid associated with site 1. The kinetics of these mutants were consistent with only site 1 being catalytically active. This was particularly striking with D297A and H299A because they disrupted hydrogen bonds between the lid and a neighbouring subunit only when in the open conformation and were distant from site 2. These observations also provided the first evidence that the flexibility and stability of lid conformations are important in catalysis. The deletion of the lid to mimic the plant oxalate oxidase led to a loss of decarboxylase activity, but only a slight elevation in the oxalate oxidase side reaction, implying other changes are required to afford a reaction specificity switch. The four mutant crystal structures (R92A, E162A, Δ162–163 and S161A) strongly support the hypothesis that site 2 is purely structural.

Evaluation of Oxalate Decarboxylase and Oxalate Oxidase for Industrial Applications

2009 ◽  
Vol 161 (1-8) ◽  
pp. 255-263 ◽  
Author(s):  
Pierre Cassland ◽  
Anders Sjöde ◽  
Sandra Winestrand ◽  
Leif J. Jönsson ◽  
Nils-Olof Nilvebrant
Keyword(s):  
Oxalate Oxidase ◽  
Industrial Applications ◽  
Oxalate Decarboxylase

scholarly journals Barley (Hordeum vulgare) oxalate oxidase is a manganese-containing enzyme

1999 ◽  
Vol 343 (1) ◽  
pp. 185-190 ◽  
Author(s):  
Laura REQUENA ◽  
Stephen BORNEMANN
Keyword(s):  
Hordeum Vulgare ◽  
Epr Spectroscopy ◽  
Active Site ◽  
Homology Model ◽  
Direct Conversion ◽  
Oxalate Oxidase ◽  
Metal Analysis ◽  
Seedling Root

Oxalate oxidase (EC 1.2.3.4) catalyses the conversion of oxalate and dioxygen into CO2 and H2O2. The barley (Hordeum vulgare) seedling root enzyme was purified to homogeneity and shown by metal analysis and EPR spectroscopy to contain Mn(II) at up to 0.80 atom per subunit. The involvement of Mn and neither flavin, Cu nor Fe in the direct conversion of dioxygen to H2O2 makes oxalate oxidase unique. A model of the active site of the holoenzyme based on a homology model of the apoenzyme is proposed.

scholarly journals Kinetic and Spectroscopic Studies of Bicupin Oxalate Oxidase and Putative Active Site Mutants

2013 ◽  
Vol 27 (S1) ◽  
Author(s):  
Ellen W Moomaw ◽  
Eric Hoffer ◽  
Patricia Moussatche ◽  
John Salerno ◽  
Morgan Grant ◽  
...  
Keyword(s):  
Active Site ◽  
Spectroscopic Studies ◽  
Oxalate Oxidase ◽  
Putative Active Site ◽  
Active Site Mutants

scholarly journals The Structure of Oxalate Decarboxylase at its Active pH

2018 ◽  
Author(s):  
M. J. Burg ◽  
J. L. Goodsell ◽  
U. T. Twahir ◽  
S. D. Bruner ◽  
A. Angerhofer
Keyword(s):  
Active Site ◽  
Density Functional ◽  
Quaternary Structure ◽  
Density Functional Theory Calculations ◽  
Strong Hydrogen Bond ◽  
Binding Modes ◽  
Oxalate Decarboxylase ◽  
Direct Role ◽  
Closed Conformation ◽  
Loop Region

AbstractOxalate decarboxylase catalyzes the redox-neutral unimolecular disproportionation reaction of oxalic acid. The pH maximum for catalysis is ~4.0 and activity is negligible above pH7. Here we report on the first crystal structure of the enzyme in its active pH range at pH4.6, and at a resolution of 1.45 Å, the highest to date. The fundamental tertiary and quaternary structure of the enzyme does not change with pH. However, the low pH crystals are heterogeneous containing both a closed and open conformation of a flexible loop region which gates access to the N-terminal active site cavity. Residue E162 in the closed conformation points away from the active-site Mn ion owing to the coordination of a buffer molecule, acetate. Since the quaternary structure of the enzyme appears unaffected by pH many conclusions drawn from the structures taken at high pH remain valid. Density functional theory calculations of the possible binding modes of oxalate to the N-terminal Mn ion demonstrate that both mono- and bi-dentate coordination modes are possible in the closed conformation with an energetic preference for the bidentate binding mode. The simulations suggest that R92 plays an important role as a guide for positioning the substrate in its catalytically competent orientation. A strong hydrogen bond is seen between the bi-dentate bound substrate and E101, one of the coordinating ligands for the N-terminal Mn ion. This suggests a more direct role of E101 as a transient base during the first step of catalysis.

scholarly journals Investigating the roles of putative active site residues in the oxalate decarboxylase from Bacillus subtilis

2007 ◽  
Vol 464 (1) ◽  
pp. 36-47 ◽  
Author(s):  
Draženka Svedružić ◽  
Yong Liu ◽  
Laurie A. Reinhardt ◽  
Ewa Wroclawska ◽  
W. Wallace Cleland ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Active Site ◽  
Active Site Residues ◽  
Oxalate Decarboxylase ◽  
Putative Active Site

scholarly journals Active site geometry of oxalate decarboxylase from Flammulina velutipes: Role of histidine-coordinated manganese in substrate recognition

2009 ◽  
Vol 11 (9) ◽  
pp. 2138-2147 ◽  
Author(s):  
Subhra Chakraborty ◽  
Niranjan Chakraborty ◽  
Deepti Jain ◽  
Dinakar M. Salunke ◽  
Asis Datta
Keyword(s):  
Active Site ◽  
Substrate Recognition ◽  
Flammulina Velutipes ◽  
Oxalate Decarboxylase

scholarly journals Kinetic and Spectroscopic Studies of Bicupin Oxalate Oxidase and Putative Active Site Mutants

PLoS ONE ◽  
2013 ◽  
Vol 8 (3) ◽  
pp. e57933 ◽  
Author(s):  
Ellen W. Moomaw ◽  
Eric Hoffer ◽  
Patricia Moussatche ◽  
John C. Salerno ◽  
Morgan Grant ◽  
...  
Keyword(s):  
Active Site ◽  
Spectroscopic Studies ◽  
Oxalate Oxidase ◽  
Putative Active Site ◽  
Active Site Mutants

CO2 Migration Pathways in Oxalate Decarboxylase and Clues about Its Active Site

2013 ◽  
Vol 117 (41) ◽  
pp. 12451-12460 ◽  
Author(s):  
Tarak Karmakar ◽  
Ganga Periyasamy ◽  
Sundaram Balasubramanian
Keyword(s):  
Active Site ◽  
Oxalate Decarboxylase ◽  
Migration Pathways

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

2007 ◽  
Vol 282 (10) ◽  
pp. 7011-7023 ◽  
Author(s):  
Mei M. Whittaker ◽  
Heng-Yen Pan ◽  
Erik T. Yukl ◽  
James W. Whittaker
Keyword(s):  
Active Site ◽  
Oxalate Oxidase ◽  
Redox Transformations ◽  
Manganese Ion ◽  
Burst Kinetics
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