title: Validation of a general method for activity estimation of cyanide evolving oxidoreductases.
abstract: 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.
authors.affiliation: Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos 62210, Mexico.
authors.forename: Francisco
authors.initials: F
authors.lastname: Gasteazoro
authors.affiliation: Institut des Sciences Moléculaires de Marseille, Aix Marseille Université, Centrale Marseille, CNRS, iSm2, UMR 7313, 13397 Marseille, France.
authors.forename: Ariane Jalila
authors.initials: AJ
authors.lastname: Simaan
authors.affiliation: Unidad de Escalamiento y Planta Piloto, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos 62210, Mexico.
authors.forename: Raunel
authors.initials: R
authors.lastname: Tinoco-Valencia
authors.affiliation: Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos 62210, Mexico. Electronic address: brenda@ibt.unam.mx.
authors.forename: Brenda
authors.initials: B
authors.lastname: Valderrama
mesh_terms: D000594:Amino Acid Oxidoreductases / Q000378:metabolism*; D057075:Enzyme Assays / Q000379:methods*; D007211:Indoles / Q000737:chemistry; D018551:Solanum lycopersicum / Q000201:enzymology; D009570:Nitriles / Q000737:chemistry / Q000378:metabolism*
chemical_list: D007211:Indoles; D009570:Nitriles; D000594:Amino Acid Oxidoreductases; C059460:1-aminocyclopropane-1-carboxylic acid oxidase
keywords: Biocatalysis; Enzymes; Ethylene; Gas chromatography; Liquid chromatography