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Related Concept Videos

Enzyme Kinetics01:19

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Enzymes speed up reactions by lowering the activation energy of the reactants. The speed at which the enzyme turns reactants into products is called the rate of reaction. Several factors impact the rate of reaction, including the number of available reactants. Enzyme kinetics is the study of how an enzyme changes the rate of a reaction.
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Inhibitors are molecules that reduce enzyme activity by binding to the enzyme. In a normally functioning cell, enzymes are regulated by a variety of inhibitors. Drugs and other toxins can also inhibit enzymes. Some inhibitors bind to the enzyme’s active site, while others inhibit enzymatic activity by binding to other sites on the protein structure.
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Enzyme-linked receptors are proteins that act as both receptor and enzyme, activating multiple intracellular signals. This is a large group of receptors that include the receptor tyrosine kinase (RTK) family. Many growth factors and hormones bind to and activate the RTKs.
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Enzyme kinetics studies the rates of biochemical reactions. Scientists monitor the reaction rates for a particular enzymatic reaction at various substrate concentrations. Additional trials with inhibitors or other molecules that affect the reaction rate may also be performed.
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Fast Enzymatic Processing of Proteins for MS Detection with a Flow-through Microreactor
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Tyrosinase immobilized enzyme reactor: development and evaluation.

Karina Bora de Oliveira1, Keylla Lençone Mischiatti1, José Domingos Fontana2

  • 1Universidade Federal do Paraná, Departamento de Farmácia, Av. Pref. Lothário Meissner, 632, CEP 80210-170 Curitiba, PR, Brazil.

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
|December 10, 2013
PubMed
Summary

Researchers developed immobilized tyrosinase enzyme reactors (tyr-IMERs) for HPLC systems. Microwave-assisted in situ immobilization on epoxy-silica yielded the most effective reactors for screening tyrosinase inhibitors.

Keywords:
Agaricus bisporusEnzyme immobilizationImmobilized enzyme reactorTyrosinase

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Area of Science:

  • Biochemistry
  • Enzyme Technology
  • Analytical Chemistry

Background:

  • Tyrosinase is a key enzyme in melanin biosynthesis and other processes.
  • Developing efficient immobilized enzyme reactors (IMERs) is crucial for enzymatic assays.
  • High-performance liquid chromatography (HPLC) systems benefit from on-line enzymatic reactors.

Purpose of the Study:

  • To prepare and compare different immobilized tyrosinase enzyme reactors (tyr-IMERs) for on-line HPLC applications.
  • To investigate the impact of various immobilization procedures on enzyme activity, kinetics, and stability.
  • To evaluate the efficacy of the developed tyr-IMERs in screening tyrosinase inhibitors.

Main Methods:

  • Tyrosinase from Agaricus bisporus was immobilized on epoxy-silica supports prepared under different conditions.
  • Both in situ and batch immobilization techniques were employed.
  • Immobilization procedures were compared based on protein and activity retention, reactor activity, kinetics, and stability.
  • Microwave-assisted synthesis was used for epoxy-silica preparation.
  • The performance of tyr-IMERs was tested against known and potential tyrosinase inhibitors.

Main Results:

  • In situ immobilization on microwave-assisted synthesized epoxy-silica resulted in the most effective tyrosinase IMERs.
  • The study compared different immobilization methods, assessing protein retention, activity, kinetics, and stability.
  • The developed tyr-IMERs demonstrated successful application in screening known and potential tyrosinase inhibitors.

Conclusions:

  • Microwave-assisted in situ immobilization is the optimal method for preparing high-performance tyrosinase IMERs.
  • The developed tyr-IMERs are suitable for on-line HPLC analysis.
  • These tyr-IMERs can be effectively utilized for the screening of tyrosinase inhibitors.