Orotidine-5′-monophosphate decarboxylase fromPseudomonas aeruginosa PAO1: Cloning, overexpression, and enzyme characterization

1994 ◽  
Vol 29 (6) ◽  
pp. 353-359 ◽  
Author(s):  
Ulrich Strych ◽  
Susanne Wohlfarth ◽  
Ulrich K. Winkler
Keyword(s):  
Enzyme Characterization ◽  
Monophosphate Decarboxylase

scholarly journals Mechanism of the Orotidine 5′-Monophosphate Decarboxylase-Catalyzed Reaction: Evidence for Substrate Destabilization,

Biochemistry ◽  
2009 ◽  
Vol 48 (24) ◽  
pp. 5518-5531 ◽  
Author(s):  
Kui K. Chan ◽  
B. McKay Wood ◽  
Alexander A. Fedorov ◽  
Elena V. Fedorov ◽  
Heidi J. Imker ◽  
...  
Keyword(s):  
Monophosphate Decarboxylase

scholarly journals FlashPlate Scintillation Proximity Assays for Characterization and Screening of DNA Polymerase, Primase, and Helicase Activities

2001 ◽  
Vol 6 (1) ◽  
pp. 39-46 ◽  
Author(s):  
David L. Earnshaw ◽  
Andrew J. Pope
Keyword(s):  
Dna Polymerase ◽  
Drug Targets ◽  
Direct Synthesis ◽  
Enzyme Characterization ◽  
Dna Helicase ◽  
Oligonucleotide Primer ◽  
Replication Proteins ◽  
Dna Primase ◽  
Wide Range ◽  
Complementary Oligonucleotide

DNA replication proteins represent a class of extremely well-established anti-infective drug targets for which improvements in assay technology are required in order to support enzyme characterization, HTS, and structure-activity relationship studies. Replication proteins are conventionally assayed using precipitation/filtration or gelbased techniques, and are not yet all suitable for conversion into homogeneous fluorescence-based formats. We have therefore developed radiometric assays for these enzymes based upon FlashPlate technology that can be applied to a wide range of targets using a common set of reagents. This approach has allowed the rapid characterization of DNA polymerase, DNA primase, and DNA helicase activities. The resultant 96-/384-well microplate assays are suitable for primary HTS, hit selectivity determination, and/or elucidating the mechanism of action of inhibitors. In all cases, biotinylated DNA oligonucleotide substrates were tethered to streptavidin-coated scintillant-embedded FlashPlate wells. Various adaptations were employed for each enzyme activity. For DNA polymerase, a short complementary oligonucleotide primer was annealed to the longer tethered oligonucleotide, and polymerization was measured by incorporation of [3H]-dNTPs onto the growing primer 3′ end. For DNA primase, direct synthesis of short oligoribonucleotides complementary to the tethered DNA strand was measured by incorporation of [3H]-rNTPs or by subsequent polymerase extension with [3H]-dNTPs from unlabeled primers. For DNA helicase, unwinding of a [33P]-labeled oligonucleotide complementary to the tethered oligonucleotide was measured. This robust and flexible system has a number of substantial advantages over conventional assay techniques for this difficult class of enzymes.

scholarly journals Catalysis by Orotidine 5′-Monophosphate Decarboxylase: Effect of 5-Fluoro and 4′-Substituents on the Decarboxylation of Two-Part Substrates

Biochemistry ◽  
2013 ◽  
Vol 52 (3) ◽  
pp. 537-546 ◽  
Author(s):  
Bogdana Goryanova ◽  
Krisztina Spong ◽  
Tina L. Amyes ◽  
John P. Richard
Keyword(s):  
Monophosphate Decarboxylase

scholarly journals Molecular Elucidation of a Urate Oxidase from Deinococcus radiodurans for Hyperuricemia and Gout Therapy

2021 ◽  
Vol 22 (11) ◽  
pp. 5611
Author(s):  
Yi-Chih Chiu ◽  
Ting-Syuan Hsu ◽  
Chen-Yu Huang ◽  
Chun-Hua Hsu
Keyword(s):  
Uric Acid ◽  
High Stability ◽  
Deinococcus Radiodurans ◽  
Catalytic Efficiency ◽  
Enzyme Characterization ◽  
Urate Oxidase ◽  
Medical Applications ◽  
Physiological Conditions ◽  
Extremophile Microorganism ◽  
Mesophilic Conditions

Urate oxidase initiates the uric acid degradation pathways and is extensively used for protein drug development for gout therapy and serum uric acid diagnosis. We first present the biochemical and structural elucidation of a urate oxidase from the extremophile microorganism Deinococcus radiodurans (DrUox). From enzyme characterization, DrUox showed optimal catalytic ability at 30 °C and pH 9.0 with high stability under physiological conditions. Only the Mg2+ ion moderately elevated its activity, which indicates the characteristic of the cofactor-free urate oxidase family. Of note, DrUox is thermostable in mesophilic conditions. It retains almost 100% activity when incubated at 25 °C and 37 °C for 24 h. In this study, we characterized a thermostable urate oxidase, DrUox with high catalytic efficiency and thermal stability, which strengthens its potential for medical applications.

Cloning, Expression, and Enzyme Characterization of an Acid Heat-Stable Phytase from Aspergillus fumigatus WY-2

2007 ◽  
Vol 55 (1) ◽  
pp. 65-70 ◽  
Author(s):  
Yan Wang ◽  
Xiaorong Gao ◽  
Qiao Su ◽  
Wei Wu ◽  
Lijia An
Keyword(s):  
Aspergillus Fumigatus ◽  
Enzyme Characterization ◽  
Heat Stable
Sign in / Sign up

Export Citation Format

Share Document