Peroxidase Chemically Attached on Polymeric Micelle and Its Reaction with Phenolic Compounds

Publication date: Available online 29 September 2017 Source:Enzyme and Microbial Technology Author(s): Moo-Yeal Lee, Parnian Bigdelou, Kyung-Jin Hong, Kyoung-Ae Kim, Yoshitsune Shinya, Toshio Kajiuchi Horseradish peroxidase was chemically modified with comb-shaped polymaleic anhydride-alt-1-tetradecene (PMA-TD) in microemulsion systems to produce surface-active peroxidase that has capability to form micellar structures in aqueous solutions and can be concentrated at liquid/liquid interfaces without unfolding of the enzyme. For chemical modification oil-in-water (O/W) and water-in-oil (W/O) microemulsion systems composed of n-butyl acetate and a buffer solution were prepared because n-butyl acetate turned out to be less detrimental to the activity of peroxidase at high degree of modification compared to other organic solvents. The modification degree of amine groups on the surface of peroxidase by maleic anhydride groups on PMA-TD was reached at equilibrium after 1h reaction at 0°C, and 42% of amine groups were modified with 7-fold amount of PMA-TD to peroxidase (wt/wt). The activity of the PMA-TD-modified peroxidase measured with 2,4-dichlorophenol at pH 7.0 was increased by approximately 2-fold compared to native peroxidase. There was no significant shift in optimum pH after modification, and optimum pH measured with 2,4-dichlorophenol was observed at pH 7.0. For all six phenolic compounds tested, there was a significant increase in the reaction efficiency with the ...
Source: Enzyme and Microbial Technology - Category: Biotechnology Source Type: research