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Spectroscopic Investigations of Lactose Oxidase and its CDs Conjugates

Grant number: 19/07692-1
Support type:Scholarships abroad - Research
Effective date (Start): January 06, 2020
Effective date (End): January 05, 2021
Field of knowledge:Engineering - Materials and Metallurgical Engineering - Nonmetallic Materials
Principal researcher:Marystela Ferreira
Grantee:Marystela Ferreira
Host: Roger Leblanc
Home Institution: Centro de Ciências e Tecnologias para a Sustentabilidade (CCTS). Universidade Federal de São Carlos (UFSCAR). Sorocaba , SP, Brazil
Research place: University of Miami, United States  

Abstract

Objective:To study Langmuir monolayer properties of lactose oxidase and CDs-Conjugate and to furtherinvestigate the interactions between the ligand on the subphase and the monolayersBackgroundSurface chemistry is an approach in biophysical chemistry that can be used to understandbiological functions such as the nature of the cell membrane interfacial, properties of variouspeptides and proteins. The best way to understand the interactions of surface proteins with theirmembrane and subphase environment is to study their air-subphase behavior by using theLangmuir film technique. Basic information like protein packing and conformation studies canbe obtained from measurement of the surface pressure, average molecular area and surfacepotential, however, detailed information can be obtained by employing additional spectroscopicand microscopic studies at different surface pressures. In Langmuir monolayer, amphiphilicmolecules are spread at the air-water interface. In doing so, the molecular packing is controlled bycompression using movable barriers. The study of Langmuir film is basically used to measurethe surface pressure as a function of surface area available to surfactant molecules at constanttemperature which is so called as surface pressure-area (À-A) isotherm. This measurement givesinformation about the existence of different phase transitions and the stability of the film atspecific temperature and pH .The isotherm behavior of the Langmuir film is usually estimated bythe physical and chemical properties of amphiphilic molecules, temperature and composition ofthe subphase.Lactose oxidase (LO) oxidizes lactose to lactobionic acid, producing H2O2 that could actas an antimicrobial and activate the lactoperoxidase system (LPS), an antimicrobial system presentin milk. As per our knowledge, the Langmuir monolayer property of this important enzyme isunknown till this date. In this project we aim to study the interfacial behavior of the LO. Further,we aim to conjugate the LO with carbon dots (CDs) and study the surface property of the conjugate.As we were interested to study the LO Langmuir monolayer at air/subphase interface, wewill utilize the Langmuir monolayer film technique to learn basic surface chemistry properties andto investigate the behavior of LO at the air-subphase interface at various surface pressures thatcorrespond to different states, ranging from the gaseous phase to the liquid compressed phase.Further, we will investigate the LO Langmuir monolayer in absence and presence of CDs ofvarying concentration on the water subphase. Moreover, the conjugation of enzyme with carbondots provides various nanohybrids with new possibilities in biosensor applications. Also, theactivity and stability of enzyme can be enhanced due to the larger surface area of nanomaterials.Methodology:To achieve our goal, we will follow four basic aims as shown below:Aim 1: Langmuir monolayer and Langmuir Blodgett (LB) studies of Lactose oxidase at differentpH and temperatures.Aim 2: Langmuir monolayer and LB studies of the CDs-Lactose oxidase conjugatesAim 3: Interactions with the ligand (lactose) in the subphase, both with Lactose oxidase andconjugates.Aim 4: Use of the conjugate to develop a biosensing platform.

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