title: Purification and Partial Characterization of β-Glucosidase in Chayote (Sechium edule).
abstract: β-Glucosidase (EC 3.2.1.21) is a prominent member of the GH1 family of glycoside hydrolases. The properties of this β-glucosidase appear to include resistance to temperature, urea, and iodoacetamide, and it is activated by 2-ME, similar to other members. β-Glucosidase from chayote (Sechium edule) was purified by ionic-interchange chromatography and molecular exclusion chromatography. Peptides detected by LC-ESI-MS/MS were compared with other β-glucosidases using the BLAST program. This enzyme is a 116 kDa protein composed of two sub-units of 58 kDa and shows homology with Cucumis sativus β-glucosidase (NCBI reference sequence XP_004154617.1), in which seven peptides were found with relative masses ranging from 874.3643 to 1587.8297. The stability of β-glucosidase depends on an initial concentration of 0.2 mg/mL of protein at pH 5.0 which decreases by 33% in a period of 30 h, and then stabilizes and is active for the next 5 days (pH 4.0 gives similar results). One hundred μg/mL β-D-glucose inhibited β-glucosidase activity by more than 50%. The enzyme had a Km of 4.88 mM with p-NPG and a Kcat of 10,000 min(-1). The optimal conditions for the enzyme require a pH of 4.0 and a temperature of 50 °C.
authors.affiliation: Unidad de Bioquímica e Inmunología, Instituto Tecnológico de Oaxaca, Oaxaca 68030, Mexico. sespindolam@hotmail.com.
authors.forename: Sergio Espíndola
authors.initials: SE
authors.lastname: Mateos
authors.affiliation: Unidad de Bioquímica e Inmunología, Instituto Tecnológico de Oaxaca, Oaxaca 68030, Mexico. carloscervantes.ox@outlook.com.
authors.forename: Carlos Alberto Matías
authors.initials: CA
authors.lastname: Cervantes
authors.affiliation: Facultad de Medicina de la, Universidad Nacional Autónoma de México, Distrito Federal 04510, Mexico. ezenteno@servidor.unam.mx.
authors.forename: Edgar
authors.initials: E
authors.lastname: Zenteno
authors.affiliation: Unité Mixte de Recherche CNRS/USTL 8576, Glycobiologie Structurale et Fonctionnelle, Université des Sciences et Technologies de Lille 1, Villeneuve d'Ascq 59655, France. Marie-Christine.Slomianny@univ-lille1.fr.
authors.forename: Marie-Christine
authors.initials: MC
authors.lastname: Slomianny
authors.affiliation: Centro de Investigación Medicina-UNAM-UABJO, Facultad de Medicina y Cirugía, Universidad Autónoma "Benito Juárez" de Oaxaca, Oaxaca 68050, Mexico. juan_alpuche@hotmail.com.
authors.forename: Juan
authors.initials: J
authors.lastname: Alpuche
authors.affiliation: Centro de Investigación Medicina-UNAM-UABJO, Facultad de Medicina y Cirugía, Universidad Autónoma "Benito Juárez" de Oaxaca, Oaxaca 68050, Mexico. fuegoblanco68@yahoo.com.mx.
authors.forename: Pedro
authors.initials: P
authors.lastname: Hernández-Cruz
authors.affiliation: Centro de Investigación Medicina-UNAM-UABJO, Facultad de Medicina y Cirugía, Universidad Autónoma "Benito Juárez" de Oaxaca, Oaxaca 68050, Mexico. rmc.azul@gmail.com.
authors.forename: Ruth
authors.initials: R
authors.lastname: Martínez-Cruz
authors.affiliation: Centro de Investigación Medicina-UNAM-UABJO, Facultad de Medicina y Cirugía, Universidad Autónoma "Benito Juárez" de Oaxaca, Oaxaca 68050, Mexico. socopina12@hotmail.com.
authors.forename: Maria
authors.initials: M
authors.lastname: del Socorro Pina Canseco
authors.affiliation: Unidad de Bioquímica e Inmunología, Instituto Tecnológico de Oaxaca, Oaxaca 68030, Mexico. perezcampos123@yahoo.es.|Centro de Investigación Medicina-UNAM-UABJO, Facultad de Medicina y Cirugía, Universidad Autónoma "Benito Juárez" de Oaxaca, Oaxaca 68050, Mexico. perezcampos123@yahoo.es.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Pérez-Campos
authors.affiliation: Unidad de Bioquímica e Inmunología, Instituto Tecnológico de Oaxaca, Oaxaca 68030, Mexico. jmsanchezrubio@gmail.com.
authors.forename: Manuel Sánchez
authors.initials: MS
authors.lastname: Rubio
authors.affiliation: Centro de Investigación Medicina-UNAM-UABJO, Facultad de Medicina y Cirugía, Universidad Autónoma "Benito Juárez" de Oaxaca, Oaxaca 68050, Mexico. laurapcm@prodigy.net.mx.
authors.forename: Laura Pérez-Campos
authors.initials: LP
authors.lastname: Mayoral
authors.affiliation: Unidad de Bioquímica e Inmunología, Instituto Tecnológico de Oaxaca, Oaxaca 68030, Mexico. martinezcu9@hotmail.com.
authors.forename: Margarito
authors.initials: M
authors.lastname: Martínez-Cruz
mesh_terms: D000595:Amino Acid Sequence; D011232:Chemical Precipitation; D002852:Chromatography, Ion Exchange; D019662:Cucurbitaceae / Q000201:enzymology*; D004795:Enzyme Stability; D005947:Glucose / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D010940:Plant Proteins; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D001617:beta-Glucosidase / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D010940:Plant Proteins; D001617:beta-Glucosidase; D005947:Glucose
keywords: Cucurbitaceae; Sechium edule; glycosyl hydrolases; β-glucosidase