Robust nanobioconjugates of Candida antarctica lipase B – Multiwalled carbon nanotubes: Characterization and application for multiple usages in non-aqueous biocatalysis

• Published in: Bioresource technology Vol. 140; pp. 103 - 110
• Main Authors: Raghavendra, Tripti, Basak, Arpana, Manocha, Lalit M., Shah, Amita R., Madamwar, Datta
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier Ltd 01.07.2013; Elsevier
• Subjects: agitation; Biocatalysis; Biological and medical sciences; Biotechnology; Candida antarctica; Candida antarctica lipase B; Carbodiimides; carbon nanotubes; chemistry; Enzymes; Enzymes, Immobilized; Enzymes, Immobilized - metabolism; Esterification; Esters; flavor; Fourier transform infrared spectroscopy; Fundamental and applied biological sciences. Psychology; Fungal Proteins; Fungal Proteins - metabolism; Immobilization; immobilized enzymes; Kinetics; Lipase; Lipase - metabolism; metabolism; methods; Multiwalled carbon nanotubes; Nanoconjugates; Nanoconjugates - chemistry; Nanoconjugates - ultrastructure; Nanostructure; Nanotechnology; Nanotechnology - methods; Nanotubes, Carbon; Nanotubes, Carbon - chemistry; Nanotubes, Carbon - ultrastructure; Non-aqueous biocatalysis; Pseudozyma antarctica; Raman spectroscopy; Recycling; Reusability; Solvents; Spectroscopy, Fourier Transform Infrared; Spectrum Analysis, Raman; succinic acid; Synthesis; Temperature; thermal analysis; Thermogravimetry; transmission electron microscopy; triacylglycerol lipase; ultrastructure; Water; Multiwalled carbon nanotubes; Candida antarctica lipase B; Non-aqueous biocatalysis; Reusability; Esterification; candida antarctica; Enzyme; Nanotube; Reuse; Carbon; Biocatalyst; Fungi; Aqueous medium; Fungi Imperfecti; Lipolytic enzyme
• ISSN: 0960-8524; 1873-2976; 1873-2976
• DOI: 10.1016/j.biortech.2013.04.071

•Covalent immobilization of CALB on MWCNTs using two chemical procedures.•Characterization of the nanoconjugates by TEM, FTIR, TGA and Raman spectroscopy.•Synthesis of pentyl valerate in organic solvents using MWCNTs–CALB conjugates.•Reusability of the bioconjugates for 50 cycles for ester synthesis. Present work elucidates two approaches for covalent attachment of the enzyme Candida antarctica lipase B (CALB) on multiwalled carbon nanotubes (MWCNTs). One method of enzyme immobilization involved carbodiimide chemistry while in the other approach, the cross linker (3-Aminopropyl) triethoxysilane (APTES) followed by succinic acid anhydride (SAA) were employed prior to carbodiimide activation. Modified MWCNTs were characterized by transmission electron microscopy (TEM), Fourier transformation infrared spectroscopic (FTIR), Raman spectroscopy and thermal gravitometric analysis (TGA). The lipase–MWCNTs conjugates were applied for synthesis of the flavor ester ‘pentyl valerate’ in cyclohexane and effects of solvent, temperature and agitation on ester synthesis were studied. Upon subject to reusability studies for 50 cycles, the bionanoconjugates were found to be highly sturdy and exhibited ∼79% activity (immobilization using carbodiimide) whereas the nanoconjugate prepared using APTES and SAA retained only up to ∼30% activity.