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    Tyrosinase enzyme Langmuir monolayer: Surface chemistry and spectroscopic study

    Year: 2020

    Journal: J. Colloid Interface Sci., Volume 564, MAR 22, page 254–263

    Authors: Paudyal, S; Sharma, SK; da Silva, RLCG; Mintz, KJ; Liyanage, PY; Al-Youbi, AO; Bashammakh, AS; El-Shahawi, MS; Leblanc, RM

    Organizations: National Science Foundation (NSF), United StatesNational Science Foundation (NSF) [GR-011298]

    Keywords: Tyrosinase; Surface chemistry; Langmuir monolayer; Infrared reflection-absorption spectroscopy; Langmuir-Blodgett film

    This study investigates the surface chemistry properties of the tyrosinase enzyme Langmuir monolayer at air-aqueous interface using sodium chloride in the subphase to induce the surface activity of the enzyme. Investigation of surface packing and stability of the tyrosinase Langmuir monolayer were performed using surface chemistry experiments while spectroscopic analysis was done to study enzyme conformation. It was found that the tyrosinase enzyme forms a fluid film at air-aqueous interface with good stability as shown by compression-decompression cycles experiments and stability measurements at various surface pressures. UV-vis absorption and fluorescence measurements at different surface pres-sures revealed that the Langmuir monolayer has good homogeneity with no evidence of aggregates during compression. To gain insight on the conformation of tyrosinase Langmuir monolayer p-polarized infrared-reflection absorption spectroscopy was used. It was found that at high surface pressures the predominant secondary structures were beta-sheets while at lower surface pressure both alpha -helices and beta-sheets were present. The circular dichroism spectra were obtained by transferring the Langmuir mono-layer at 10 mN.m(-1) to a solid quartz support (Langmuir-Blodgett film, LB film), which showed that the major conformation present were alpha-helices. Images from the immobilized LB films were obtained using atomic force microscopy which showed homogenous and regular deposition with a mean thickness ranging from 3 to 4 nm. (C) 2019 Elsevier Inc. All rights reserved.
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