title: Controlling the molecular architecture of lactase immobilized in Langmuir-Blodgett films of phospholipids to modulate the enzyme activity.
abstract: In this present work, the adsorption of the enzyme lactase onto Langmuir monolayers of the phospholipid dimyristoylphosphatidic acid (DMPA) was investigated and characterized with surface pressure-area isotherms, surface potential-area isotherms and polarization-modulated infrared reflection-absorption spectroscopy (PM-IRRAS). The adsorption of the enzyme at the air-water interface expanded the lipid monolayer and increased the film compressibility at high surface pressures. Amide bands in the PM-IRRAS spectra were identified, with the CN and CO dipole moments lying parallel to the monolayer plane, revealing that the structuring of the enzyme into β-sheets was kept in the mixed monolayer. The enzyme-lipid films were transferred from the floating monolayer to solid supports as Langmuir-Blodgett (LB) films and characterized with fluorescence spectroscopy and atomic force microscopy. The catalytic activity of the films was measured and compared to the homogenous medium. The enzyme accommodated in the LB films preserved more than 80% of the enzyme activity after 20days, in contrast for the homogeneous medium, which preserved less than 60% of the enzyme activity. The method presented in this present work not only allows for an enhanced catalytic activity toward lactose, but also can help explain why certain film architectures exhibit better performance.
authors.affiliation: Institute of Environmental, Chemical and Pharmaceutical Sciences, Federal University of Sao Paulo, Brazil.
authors.forename: Fábio de Paula
authors.initials: FP
authors.lastname: Ayoub
authors.affiliation: Institute of Environmental, Chemical and Pharmaceutical Sciences, Federal University of Sao Paulo, Brazil. Electronic address: lcaseli@gmail.com.
authors.forename: Luciano
authors.initials: L
authors.lastname: Caseli
mesh_terms: D000327:Adsorption; D000814:Aniline Compounds / Q000737:chemistry; D002384:Catalysis; D004548:Elasticity; D004800:Enzymes, Immobilized / Q000737:chemistry*; D043322:Lactase / Q000737:chemistry*; D008055:Lipids / Q000737:chemistry; D018625:Microscopy, Atomic Force; D010743:Phospholipids / Q000737:chemistry*; D011312:Pressure; D013050:Spectrometry, Fluorescence; D013055:Spectrophotometry, Infrared; D013499:Surface Properties; D013696:Temperature; D014867:Water / Q000737:chemistry
chemical_list: D000814:Aniline Compounds; D004800:Enzymes, Immobilized; D008055:Lipids; D010743:Phospholipids; D014867:Water; C050795:N,N-dimethyl-4-anisidine; D043322:Lactase
keywords: Biosensors; Enzyme activity; Lactase; Langmuir monolayers; Langmuir-Blodgett films