scholarly journals Pseudomonas aeruginosaC5-Mannuronan Epimerase:  Steady-State Kinetics and Characterization of the Product†

Biochemistry ◽  
2006 ◽  
Vol 45 (2) ◽  
pp. 552-560 ◽  
Author(s):  
Agoston Jerga ◽  
Aniruddha Raychaudhuri ◽  
Peter A. Tipton
Keyword(s):  
Steady State ◽  
Steady State Kinetics

Steady-State Kinetics and Spectroscopic Characterization of Enzyme–tRNA Interactions for the Non-Heme Diiron tRNA-Monooxygenase, MiaE

Biochemistry ◽  
2014 ◽  
Vol 54 (2) ◽  
pp. 363-376 ◽  
Author(s):  
Bishnu P. Subedi ◽  
Andra L. Corder ◽  
Siai Zhang ◽  
Frank W. Foss ◽  
Brad S. Pierce
Keyword(s):  
Steady State ◽  
Spectroscopic Characterization ◽  
Steady State Kinetics

Characterization of recombinant human acetyl-CoA carboxylase-2 steady-state kinetics

2009 ◽  
Vol 1794 (6) ◽  
pp. 961-967 ◽  
Author(s):  
Virendar K. Kaushik ◽  
Michael Kavana ◽  
Jessica M. Volz ◽  
Stephen C. Weldon ◽  
Susan Hanrahan ◽  
...  
Keyword(s):  
Steady State ◽  
Acetyl Coa Carboxylase ◽  
Acetyl Coa ◽  
Steady State Kinetics

scholarly journals Purification and characterization of two glutathione S-aryltransferase activities from rat liver.

1975 ◽  
Vol 147 (3) ◽  
pp. 513-522 ◽  
Author(s):  
P Askelöf ◽  
C Guthenberg ◽  
I Jakobson ◽  
B Mannervik
Keyword(s):  
Steady State ◽  
Rat Liver ◽  
Substrate Concentration ◽  
Thiol Group ◽  
Purification And Characterization ◽  
Molecular Weights ◽  
Enzymic Reaction ◽  
Steady State Kinetics

Two forms of glutathione S-aryltransferase were purified from rat liver. The only differences noted between the two forms were in the chromatographic and electrophoretic properties, which permitted the separation of the two species. The molecular weights of the enzyme and its subunits were estimated as about 50000 and 23000 respectively. The steady-state kinetics did no follow Michaelis-Menten kinetics when one substrate concentration was kept constant while the second substrate concentration was varied. Several S-substituted GSH derivatives were tested as inhibitors of the enzymic reaction. The enzyme was inactivated by thiol-group reagents.

scholarly journals Characterization of Neurospora crassa catabolic dehydroquinase purified from N. crassa and Escherichia coli

1982 ◽  
Vol 203 (3) ◽  
pp. 769-773 ◽  
Author(s):  
A R Hawkins ◽  
W R Reinert ◽  
N H Giles
Keyword(s):  
Escherichia Coli ◽  
Steady State ◽  
Neurospora Crassa ◽  
Protein Sequence ◽  
Sequence Data ◽  
E Coli ◽  
Steady State Kinetics ◽  
Electrophoretic Data ◽  
Protein Sequence Data

1. Neurospora crassa catabolic dehydroquinase has been purified from N. crassa and Escherichia coli. 2. Protein-sequence and gel-electrophoretic data show that apparently pure, homogeneous native dehydroquinase is a mixture of intact and proteinase-cleaved enzyme monomers. 3. Protein-sequence data and steady-state kinetics show that the catabolic dehydroquinase gene of N. crassa is expressed with fidelity in E. coli.

Characterization of the PLP-dependent transaminase initiating azasugar biosynthesis

2018 ◽  
Vol 475 (13) ◽  
pp. 2241-2256
Author(s):  
Jeffrey M. Arciola ◽  
Nicole A. Horenstein
Keyword(s):  
Steady State ◽  
Specific Activity ◽  
Paenibacillus Polymyxa ◽  
Binding Pocket ◽  
Kinetic Mechanism ◽  
Steady State Kinetics ◽  
Ping Pong ◽  
Amino Donor ◽  
Event Based

Biosynthesis of the azasugar 1-deoxynojirimycin (DNJ) critically involves a transamination in the first committed step. Here, we identify the azasugar biosynthetic cluster signature in Paenibacillus polymyxa SC2 (Ppo), homologous to that reported in Bacillus amyloliquefaciens FZB42 (Bam), and report the characterization of the aminotransferase GabT1 (named from Bam). GabT1 from Ppo exhibits a specific activity of 4.9 nmol/min/mg at 30°C (pH 7.5), a somewhat promiscuous amino donor selectivity, and curvilinear steady-state kinetics that do not reflect the predicted ping-pong behavior typical of aminotransferases. Analysis of the first half reaction with l-glutamate in the absence of the acceptor fructose 6-phosphate revealed that it was capable of catalyzing multiple turnovers of glutamate. Kinetic modeling of steady-state initial velocity data was consistent with a novel hybrid branching kinetic mechanism which included dissociation of PMP after the first half reaction to generate the apoenzyme which could bind PLP for another catalytic deamination event. Based on comparative sequence analyses, we identified an uncommon His-Val dyad in the PLP-binding pocket which we hypothesized was responsible for the unusual kinetics. Restoration of the conserved PLP-binding site motif via the mutant H119F restored classic ping-pong kinetic behavior.

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