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Cofactors and Coenzymes01:24

Cofactors and Coenzymes

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Enzymes are proteins made of amino acids. The functional group of each constituent amino acid catalyzes a wide variety of chemical reactions via ionic interactions or acid-base reactions. However, amino acids cannot catalyze oxidation-reduction and group transfer reactions and need to be aided by non-protein components called cofactors. Cofactors are also referred to as the chemical teeth of an enzyme.
Cofactors can be metallic ions or organic molecules called coenzymes. These types of helper...
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Cofactors and Coenzymes01:27

Cofactors and Coenzymes

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Cofactors and Coenzymes01:27

Cofactors and Coenzymes

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Enzymes require additional components for proper function. There are two such classes of molecules: cofactors and coenzymes. Cofactors are metallic ions and coenzymes are non-protein organic molecules. Both of these types of helper molecule can be tightly bound to the enzyme or bound only when the substrate binds.
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Enzyme Kinetics01:19

Enzyme Kinetics

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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.
Scientists typically study enzyme kinetics with a fixed amount of enzyme in the controlled environment of a test tube. When more reactant, or substrate, is...
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Catalytically Perfect Enzymes01:07

Catalytically Perfect Enzymes

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The theory of catalytically perfect enzymes was first proposed by W.J. Albery and J. R. Knowles in 1976. These enzymes catalyze biochemical reactions at high-speed. Their catalytic efficiency values range from 108-109 M-1s-1. These enzymes are also called 'diffusion-controlled' as the only rate-limiting step in the catalysis is that of the substrate diffusion into the active site. Examples include triose phosphate isomerase, fumarase, and superoxide dismutase.
 
Most enzymes...
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Enzymes02:34

Enzymes

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Inside living organisms, enzymes act as catalysts for many biochemical reactions involved in cellular metabolism. The role of enzymes is to reduce the activation energies of biochemical reactions by forming complexes with its substrates. The lowering of activation energies favor an increase in the rates of biochemical reactions.
Enzyme deficiencies can often translate into life-threatening diseases. For example, a genetic abnormality resulting in the deficiency of the enzyme G6PD...
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Updated: Mar 15, 2026

Expression and Purification of Nuclease-Free Oxygen Scavenger Protocatechuate 3,4-Dioxygenase
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Expression and Purification of Nuclease-Free Oxygen Scavenger Protocatechuate 3,4-Dioxygenase

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Un par de enzimas y cofactores ortogonales evolucionados

Evan W Reynolds1, Matthew W McHenry1, Fabien Cannac1

  • 1Department of Chemistry, University of North Carolina-Chapel Hill , 125 South Road, CB 3290, Chapel Hill, North Carolina 27599, United States.

Journal of the American Chemical Society
|August 31, 2016
PubMed
Resumen
Este resumen es generado por máquina.

Los investigadores desarrollaron una estrategia ortogonal de pares de enzimas y hemo para expandir la funcionalidad de las hemoproteínas. Un citocromo P450 modificado utiliza selectivamente un cofactor no natural, lo que permite nuevas reacciones químicas.

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Área de la Ciencia:

  • La bioquímica
  • Biotecnología
  • Ingeniería de enzimas

Sus antecedentes:

  • Las hemoproteínas son enzimas cruciales con cofactores hemo.
  • La expansión de la funcionalidad de las hemoproteínas requiere nuevas estrategias de incorporación de cofactores.
  • Los citocromo P450 son enzimas versátiles a menudo diseñadas para nuevas funciones.

Objetivo del estudio:

  • Desarrollar una estrategia ortogonal de pares enzima/hemo para expandir la funcionalidad de las hemoproteínas.
  • Diseñar una enzima del citocromo P450 (P450BM3) para la incorporación selectiva de un derivado del hemo no nativo.
  • Demostrar la utilidad del par enzima/cofactor diseñado en la catálisis.

Principales métodos:

  • Aprovechó la proteína de transporte de hemo ChuA para importar derivados de hemo.
  • P450BM3 evolucionado para la unión selectiva de deuteroporfirina IX de hierro (Fe-DPIX) con el hemo endógeno.
  • Se utilizó la cristalografía de rayos X para dilucidar la base estructural de la selectividad del cofactor.
  • Se ha probado la actividad catalítica del par ortogonal en ciclopropanación mediada por carbenoides.

Principales resultados:

  • El P450BM3 modificado incorporó selectivamente Fe-DPIX en presencia de hemo nativo.
  • El análisis estructural reveló mutaciones que confieren selectividad a través de interacciones estéricas y de unión.
  • El par enzima/cofactor evolucionado demostró actividad en la ciclopropanación no natural de las olefinas.
  • Se ha generado con éxito un par ortogonal enzima/cofactor.

Conclusiones:

  • La estrategia desarrollada permite la creación de pares ortogonales de enzimas y cofactores.
  • Este enfoque amplía significativamente la diversidad de cofactores para las metalenzimas artificiales.
  • El sistema diseñado ofrece una plataforma para nuevas aplicaciones biocatalíticas.
  • La metodología promete aplicaciones más amplias en la ingeniería enzimática y la biología sintética.