Immobilization of a beta-glucosidase and an endoglucanase in ferromagnetic nanoparticles: A study of synergistic effects

Author(s)

Carli, Sibeli ^[1] ; Buffoni de Campos Carneiro, Lara Aparecida ^[2] ; Ward, Richard John ^[1] ; Meleiro, Luana Parras ^[1]

Affiliation

^[1] Univ Sao Paulo, Fac Filosofia Ciencias & Letras Ribeirao Preto, Dept Quim, BR-14040901 Ribeirao Preto, SP - Brazil

^[2] Univ Sao Paulo, Fac Med Ribeirao Preto, Dept Bioquim & Imunol, BR-14049900 Ribeirao Preto, SP - Brazil

• Document type: Journal article
• Source: Protein Expression and Purification; v. 160, p. 28-35, AUG 2019.
• Web of Science Citations: 2

Abstract

Nanoparticles can act as support materials for enzymatic immobilization, introducing a balance of characteristics that modulate the efficiency of biocatalysts, such as specific surface area, resistance to mass transfer and effective enzymatic loading. Magnetic nanoparticles can be easily separated using an external magnetic field, and in this work two recombinant enzymes, the beta-glucosidase from Humicola insolens (Bglhi) and the endoglucanase from Scytalidium thermophilwn (Egst) were immobilized on synthesized Fe3O4 nanoparticles derivatized with chitosan/glutaraldehyde/N-(5-amino-1-carboxy-pentyl) iminodiacetic acid and functionalized with NiCl2. The immobilization yields were about 20% for Bglhi and Egst with efficiencies of 132% and 115%, respectively. The two enzymes were also co-immobilized with yield was about 49%. The optimal temperatures of the immobilized enzymes were 70 degrees C and 55 degrees C for Egst and Bglhi, respectively. Egst hydrolyzed CMC in the presence of 4 mM MnCl2 with V-max = 625.0 +/- 6.7 U mg(-1) and K-M = 6.4 +/- 0.5 mg mL(-1) resulting in a catalytic efficiency (k(cat)/K-M) of 107.4 +/- 5.4 mg(-1) s(-1) mL. Bglhi hydrolyzed pNP-Glc with V-max = 52.7 +/- 2.7 U mg(-1) and K-M = 0.23 0.01 mM resulting in a catalytic efficiency (k(cat)/K-M) of 214.3 +/- 10.25(-1) mM(-1). The individually immobilized enzymes when combined showed a synergistic effect on the substrates tested and a very similar action when compared to the co-immobilized enzymes, suggesting excellent potential for application in biotechnological processes. (AU)

• FAPESP's process: 17/13734-3 - Engineering of an endopolygalacturonase by insertion of different Carbohydrate Binding Modules (CBMs): chimerogenesis and proximity effects
• Grantee: Sibeli de Carli
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 16/17582-0 - Thermostability modulation of a ß-glucosidase stimulated by glucose and xylose using site directed glycosylation
• Grantee: Luana Parras Meleiro Garcia
• Support Opportunities: Scholarships in Brazil - Post-Doctoral