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Validation of a general method for activity estimation of cyanide evolving oxidoreductases.
Gasteazoro, Francisco; Simaan, Ariane Jalila; Tinoco-Valencia, Raunel; Valderrama, Brenda.
Afiliación
  • Gasteazoro F; Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos 62210, Mexico.
  • Simaan AJ; Institut des Sciences Moléculaires de Marseille, Aix Marseille Université, Centrale Marseille, CNRS, iSm2, UMR 7313, 13397 Marseille, France.
  • Tinoco-Valencia R; Unidad de Escalamiento y Planta Piloto, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos 62210, Mexico.
  • Valderrama B; Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos 62210, Mexico. Electronic address: brenda@ibt.unam.mx.
Anal Biochem ; 471: 44-50, 2015 Feb 15.
Article en En | MEDLINE | ID: mdl-25447496
Ethylene is a key molecule in organic synthesis currently produced by steam cracking of fossil hydrocarbons. In nature, ethylene is produced in higher plants by 1-aminocyclopropane-1-carboxylic acid oxidase (ACCO). Biocatalytic alternatives for ethylene production are still far from being competitive with traditional production plants. Furthermore, data dispersion shown in the literature adds uncertainty to the introduction of ACCO as a biocatalyst, especially when larger numbers of isoforms or mutants are to be compared. Here we propose a new method for measuring ACCO activity based on cyanide detection. Data provided here indicate that cyanide detection is more precise, more responsive, and much more stable than any other method tested for ACCO activity estimation so far. Briefly, enzymatically produced cyanide can be detected by its derivatization with naphthalene-2,3-dicarboxyaldehide (NDA) to generate 1-cyanobenz[f]isoindole (CBI), which is further detected by high-performance liquid chromatography (HPLC) coupled with a fluorescence detector. Cyanide can be detected in the range between 0.99 and 60.17pmol, which is three orders of magnitude more sensitive than the currently used ethylene estimation method.
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Texto completo: 1 Base de datos: MEDLINE Asunto principal: Pruebas de Enzimas / Aminoácido Oxidorreductasas / Nitrilos Idioma: En Revista: Anal Biochem Año: 2015 Tipo del documento: Article País de afiliación: México

Texto completo: 1 Base de datos: MEDLINE Asunto principal: Pruebas de Enzimas / Aminoácido Oxidorreductasas / Nitrilos Idioma: En Revista: Anal Biochem Año: 2015 Tipo del documento: Article País de afiliación: México