Your Step-By-Step Guide To Measure Enzyme Activity

For Oxidoreductases Using Spectrophotometric Assays

--- published by Creative Enzymes

• Oxidoreductases
Oxidoreductases are a class of enzymes that catalyze the transfer
of electrons from one molecule to another. The molecule that gives
the electrons is called the electron donor or reductant, and the one
receiving the electrons are called the electron acceptor or oxidant.
Oxidoreductase enzymes play an important role in metabolic
process. They can be found in both aerobic and anaerobic
pathways, including glycolysis, TCA cycle, oxidative
phosphorylation, and amino acid metabolism.
To complete their functions, the oxidoreductase enzymes utilize
redox cofactors. Common redox cofactors include FAD (flavin
adenine dinucleotide), Fd (ferredoxin) FMN (flavin mononucleotide),
NAD (nicotinamide adenine dinucleotide), NADP (nicotinamide
adenine dinucleotide phosphate), Coenzyme B, Coenzyme Q, etc.
For example, the enzyme glyceraldehydes-3-phosphate
dehydrogenase reduces NAD+ to NADH in glycolysis. While NADH
is re-oxidized to NAD+ in the oxidative phosphorylation pathway to
maintain the proper re-dox state of the cell.
Pyruvate is produced during glycolysis, which is then reduced to
lactate through anaerobic glycolysis using NADH as the reductant.
The redox cofactors bear one or more aromatic or unsaturated
cyclic chemical groups in their structures. As a result, the cofactors
strongly absorb UV or visible light in the reduced or oxidized form,
or both. This makes the redox cofactors perfect reporters to the
spectrophotometric enzyme assays.
Oxidoreductases can be categorized into different subtypes,
including oxidases and dehydrogenases. Oxidase enzymes use the
molecular oxygen as the acceptor of hydrogen or electrons. To the
contrary, dehydrogenase enzymes transfer hydrogen to NAD,
NADP, or a flavin in order to oxidize a substrate. Other
oxidoreductases include reductases, peroxidases (e.g.
Lactoperoxidase), hydroxylases, and oxygenases.
Reductases could also be oxidases since most redox reactions are
reversible. The direction of the redox reaction depends on both
thermodynamics and kinetics. Peroxidases catalyze the
decomposition of hydrogen peroxide, which is a natural defensive
process against environmental invaders. Hydroxylases incorporate
hydroxyl groups into organic compounds, while oxygenases add
oxygen from molecular oxygen to the substrates.
Creative Enzymes proudly offers spectrophotometric enzyme
assays in quantification of activity levels for all types of
oxidoreductase enzymes:
• Alcohol oxidoreductases (EC 1.1, oxidoreductases that act on
the CH-OH group of donors)
• Oxidoreductases acting on the aldehyde or oxo group of donors
(EC 1.2)
• CH–CH oxidoreductases (EC 1.3, oxidoreductases that act on
the CH-CH group of donors)
• Oxidoreductases that act on the CH-NH2 group of donors (EC
1.4, amino acid oxidoreductases, monoamine oxidases)
• Oxidoreductases that act on CH-NH group of donors (EC 1.5)
• Oxidoreductases that act on NADH or NADPH (EC 1.6)
• Oxidoreductases that act on other nitrogenous compounds as
donors (EC 1.7)
• Oxidoreductases that act on a sulfur group of donors (EC 1.8)
• Oxidoreductases that act on a heme group of donors (EC 1.9)
• Oxidoreductases that act on diphenols and related substances
as donors (EC 1.10)
• Peroxidases (EC 1.11, oxidoreductases that act on peroxide as
an acceptor such as Manganese Peroxidase)
• Oxidoreductases that act on hydrogen as donors (EC 1.12)
• Oxygenases (EC 1.13), oxidoreductases that act on single
donors with incorporation of molecular oxygen
• Oxidoreductases that act on paired donors with incorporation of
molecular oxygen (EC 1.14)
• Oxidoreductases that act on superoxide radicals as acceptors
(EC 1.15)
• Oxidoreductases that oxidize metal ions (EC 1.16)
• Oxidoreductases that act on CH or CH2 groups (EC 1.17)
• Oxidoreductases that act on iron-sulfur proteins as donors (EC
1.18)
• Oxidoreductases that act on reduced flavodoxin as a donor (EC
1.19)
• Oxidoreductases that act on phosphorus or arsenic in donors
(EC 1.20)
• Oxidoreductases that act on X-H and Y-H to form an X-Y bond
(EC 1.21)
• Other oxidoreductases (EC 1.97)
As a large and important family of enzymes, oxidoreductases are
used widely in the pharmaceutical industry for syntheses of amino
acids, steroids, and other molecules; in the chemical industry for
production of specialty chemicals; in clinical diagnosis and other
analytical applications; and in other applications such as material
modification and pollution control.
For more details, please kindly click:
https://www.creative-enzymes.com/
Thanks!