Biochemical and physical characterization of the active FAD-containing form of nitroalkane oxidase from Fusarium oxysporum

Biochemistry
G Gadda, Paul F Fitzpatrick

Abstract

Nitroalkane oxidase from Fusarium oxysporum catalyzes the oxidation of nitroalkanes to aldehydes with production of nitrite and hydrogen peroxide. The enzyme has a molecular weight of 47 955 +/- 39, as determined by MALDI-TOF mass spectrometry; under nondenaturing conditions, the aggregation state of the enzyme is best described by a tetramer-dimer self-associating model, with an association constant of (8.5 +/- 4.4) x 10(6) M-1 (pH 7.0 and 4 degreesC). The amino acid composition and the N-terminal amino acid sequence do not match any known protein or open reading frame. The inactive 5-nitrobutyl-1,5-dihydroflavin found in the enzyme as purified was converted to FAD, allowing characterization of the active FAD-containing enzyme. With nitroethane as substrate, the Vmax and Km values are 655 +/- 45 min-1 and 2.9 +/- 0.5 mM at pH 8.0 and 30 degreesC, respectively. One mole of FAD per mole of monomer enzyme is required for catalysis. No activity can be detected with amino acids or alpha-hydroxy acids as substrates. Reversible removal of the FAD cofactor yields inactive enzyme. The properties of the FAD cofactor in nitroalkane oxidase are within the range described for other oxidases. The UV-visible absorbance spectrum of the active...Continue Reading

Citations

May 21, 2010·Biochemistry·José R TormosPaul F Fitzpatrick
Mar 7, 2009·Biochemistry·Annie HérouxAllen M Orville
Feb 17, 2005·Journal of the American Chemical Society·Michael P ValleyPaul F Fitzpatrick
May 20, 2004·Journal of the American Chemical Society·Michael P Valley, Paul F Fitzpatrick
Jul 24, 2004·Acta Crystallographica. Section D, Biological Crystallography·Akanksha NagpalAllen M Orville
Sep 4, 2008·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Vicki A BamfordKimberly A Watson
Dec 4, 2012·International Journal of Molecular Sciences·Pablo Sobrado
Feb 28, 2002·Proceedings of the National Academy of Sciences of the United States of America·S Colette DaubnerPaul F Fitzpatrick
Jan 9, 2010·Bioorganic Chemistry·Michael P ValleyPaul F Fitzpatrick
Jul 7, 2009·Archives of Biochemistry and Biophysics·Giovanni Gadda, Kevin Francis
May 15, 2009·The FEBS Journal·Dominic P H M HeutsMarco W Fraaije
Dec 8, 2004·Archives of Biochemistry and Biophysics·Paul F FitzpatrickMichael P Valley
Mar 9, 1999·Archives of Biochemistry and Biophysics·G Gadda, P F Fitzpatrick
Dec 10, 1999·The Journal of Biological Chemistry·M W FraaijeA Mattevi
May 9, 2001·Journal of Enzyme Inhibition·G GaddaP F Fitzpatrick
Mar 19, 2008·Journal of the American Chemical Society·Steven C RothmanC Dale Poulter
Apr 20, 2001·Accounts of Chemical Research·P F Fitzpatrick

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.