Purification and Partial Characterization of β-Glucosidase in Chayote (Sechium edule)
Abstractβ-Glucosidase (EC 220.127.116.11) 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. View Full-Text
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Mateos, S.E.; Cervantes, C.A.M.; Zenteno, E.; Slomianny, M.-C.; Alpuche, J.; Hernández-Cruz, P.; Martínez-Cruz, R.; Canseco, M.S.P.; Pérez-Campos, E.; Rubio, M.S.; Mayoral, L.P.-C.; Martínez-Cruz, M. Purification and Partial Characterization of β-Glucosidase in Chayote (Sechium edule). Molecules 2015, 20, 19372-19392.
Mateos SE, Cervantes CAM, Zenteno E, Slomianny M-C, Alpuche J, Hernández-Cruz P, Martínez-Cruz R, Canseco MSP, Pérez-Campos E, Rubio MS, Mayoral LP-C, Martínez-Cruz M. Purification and Partial Characterization of β-Glucosidase in Chayote (Sechium edule). Molecules. 2015; 20(10):19372-19392.Chicago/Turabian Style
Mateos, Sergio E.; Cervantes, Carlos A.M.; Zenteno, Edgar; Slomianny, Marie-Christine; Alpuche, Juan; Hernández-Cruz, Pedro; Martínez-Cruz, Ruth; Canseco, Maria S.P.; Pérez-Campos, Eduardo; Rubio, Manuel S.; Mayoral, Laura P.-C.; Martínez-Cruz, Margarito. 2015. "Purification and Partial Characterization of β-Glucosidase in Chayote (Sechium edule)." Molecules 20, no. 10: 19372-19392.