title: Noncovalent and covalent immobilization of oxygenase on single-walled carbon nanotube for enzymatic decomposition of aromatic hydrocarbon intermediates.
abstract: The decomposition of various aromatic hydrocarbon intermediates was examined using a recombinant oxidative enzyme immobilized on single-walled carbon nanotubes (SWCNTs). Hydroxyquinol 1,2-dioxygenase (CphA-I), which catalyzes ring cleavage of catechol and its analogues, was obtained from Arthrobacter chlorophenolicus A6 via cloning, overexpression, and subsequent purification. This recombinant enzyme was immobilized on SWCNTs by physical adsorption and covalent coupling in the absence and presence of N-hydroxysuccinimide. The immobilization yield was as high as 52.1%, and a high level of enzyme activity of up to 64.7% was preserved after immobilization. Kinetic analysis showed that the substrate utilization rates (vmax) and catalytic efficiencies (kcat/KM) of the immobilized enzyme for all substrates evaluated were similar to those of the free enzyme, indicating minimal loss of enzyme activity during immobilization. The immobilized enzyme was more stable toward extreme pH, temperature, and ionic strength conditions than the free enzyme. Thus, the oxidative enzyme immobilized on SWCNTs can be used as an effective and stable biocatalyst for the biochemical remediation process if further investigations would be carried out under field conditions.
authors: Department of Advanced Technology Fusion, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul, 143-701, Korea.|Department of Environmental Engineering, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul, 143-701, Korea.
authors: Yanasinee
authors: Y
authors: Suma
authors: Department of Environmental Engineering, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul, 143-701, Korea.
authors: Christina S
authors: CS
authors: Kang
authors: Department of Environmental Engineering, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul, 143-701, Korea. hankim@konkuk.ac.kr.
authors: Han S
authors: HS
authors: Kim
mesh_terms: D000327:Adsorption; D001173:Arthrobacter / Q000378:metabolism; D001673:Biodegradation, Environmental; D049308:Dioxygenases / Q000378:metabolism; D004800:Enzymes, Immobilized / Q000378:metabolism*; D006841:Hydrocarbons, Aromatic / Q000378:metabolism*; D037742:Nanotubes, Carbon / Q000737:chemistry; D010105:Oxygenases / Q000378:metabolism*; D013388:Succinimides; D013696:Temperature
chemical_list: D004800:Enzymes, Immobilized; D006841:Hydrocarbons, Aromatic; D037742:Nanotubes, Carbon; D013388:Succinimides; D010105:Oxygenases; D049308:Dioxygenases; C041894:hydroxyquinol 1,2-dioxygenase; C001426:N-hydroxysuccinimide
keywords: Covalent coupling; Enzyme immobilization; Physical adsorption; Recombinant enzyme; Single-walled carbon nanotubes
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