Enzymatic degradation of aromatic hydrocarbon intermediates using a recombinant dioxygenase immobilized onto surfactant-activated carbon nanotube

• Published in: Bioresource technology 2016-06, Vol.210, p.117-122
• Main Authors: Suma, Yanasinee, Lim, Heejun, Kwean, Oh Sung, Cho, Suyeon, Yang, Junwon, Kim, Yohan, Kang, Christina S., Kim, Han S.
• Format: Article
• Language: English
• Subjects: Arthrobacter; Arthrobacter - enzymology; Charcoal - pharmacology; Dioxygenase; Dioxygenases - metabolism; Enzyme immobilization; Enzyme Stability - drug effects; Enzymes, Immobilized - metabolism; Hydrocarbons, Aromatic - metabolism; Hydrogen-Ion Concentration; Kinetics; Nanotubes, Carbon - chemistry; Osmolar Concentration; Recombinant Proteins - metabolism; Single-walled carbon nanotube; Solutions; Stability; Surface-Active Agents - pharmacology; Surfactant; Temperature
• ISSN: 0960-8524; 1873-2976
• DOI: 10.1016/j.biortech.2016.01.018

[Display omitted] •Various aromatic hydrocarbon intermediates can be decomposed by enzymes effectively.•SWCNTs activated with nonionic surfactants can serve as an effective enzyme support.•Loss of enzyme activity was minimal during enzyme immobilization.•Enzyme immobilization improved the enzyme stability significantly. This study examined the enzymatic decomposition of aromatic hydrocarbon intermediates (catechol, 4-chlorocatechol, and 3-methylcatechol) using a dioxygenase immobilized onto single-walled carbon nanotube (SWCNT). The surfaces of SWCNTs were activated with surfactants. The dioxygenase was obtained by recombinant technique: the corresponding gene was cloned from Arthrobacter chlorophenolicus A6, and the enzyme was overexpressed and purified subsequently. The enzyme immobilization yield was 62%, and the high level of enzyme activity was preserved (60–79%) after enzyme immobilization. Kinetic analyses showed that the substrate utilization rates and the catalytic efficiencies of the immobilized enzyme for all substrates (target aromatic hydrocarbon intermediates) tested were similar to those of the free enzyme, indicating that the loss of enzyme activity was minimal during enzyme immobilization. The immobilized enzyme was more stable than the free enzyme against abrupt changes in pH, temperature, and ionic strength. Moreover, it retained high enzyme activity even after repetitive use.