Development and Characterization of a Novel Antibacterial Material Based on GOx Immobilized in a PVA Film
The development and characterization of a novel antibacterial material based on glucose oxidase (GOx) immobilized in a polyvinyl alcohol (PVA) film was proposed. This system acts in the presence of glucose to generate by-products as oxygen species (H2O2,
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1
Introduction
Bacterial infection is one of the most severe concerns for several research fields such as biotechnology, pharmaceutical, textiles, food packaging and storage, shoe industry, water purification, medical devices, and dental surgery equipment [1, 2]. Recently, antimicrobial agents have gained considerable interest from both an industrial and research point of view because of their potential to provide safety benefits to a diverse range of materials. Nowadays scientists need to carry out materials with a surface that has a very broad spectrum of biocide activity, that can be used repeatedly and that kills via a mechanism which will not result in the emergence of resistant strains, and that can be used for development of devices for biomedical applications [3]. This system acts in presence of glucose to generate by-products as oxygen species (H2O2, ·O2−, OH) that are well-known endogenous and exogenous toxic products for microbes in vivo [4].
2 2.1
Experimental Section Reagents and Apparatus
Glucose oxidase (type VII from Aspergillus niger; 174,000 units/g), β-d-glucose, and hydrogen peroxide 30 % were obtained from Sigma. Na2HPO4, NaH2PO4, and poly(vinyl alcohol) film (PVA, product number Z300381) were purchased from M.R. Guascito (*) • D. Chirizzi • L. Giotta Università del Salento, via Monteroni, 73100 Lecce, Italy e-mail: [email protected] L. Stabili Istituto per l’Ambiente Marino Costiero CNR, via Roma 3, 74100 Taranto, Italy C. Di Natale et al. (eds.), Sensors and Microsystems: Proceedings of the 17th National Conference, Brescia, Italy, 5-7 February 2013, Lecture Notes in Electrical Engineering 268, DOI 10.1007/978-3-319-00684-0_36, © Springer International Publishing Switzerland 2014
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Aldrich. Micrococcus lysodeikticus cells were obtained from Sigma. All reagents were analytical grade. Phosphate buffer solution (PB) pH = 7.0, I = 0.2. 1 M stock glucose solutions were allowed to mutarotate at room temperature overnight before use. XPS analysis was carried out using a Leybold LHS10 spectrometer equipped with an unmonochromatized AlKα source, and a SPECS multichannel detector and UV–vis measurements were carried out with a Cary 50 spectrophotometer (Varian). Mid-infrared spectra were acquired with a PerkinElmer Spectrum One FTIR spectrometer equipped with a deuterated triglycine sulfate (DTGS) detector.
2.2
Preparation of Antibacterial Composite Material
The PVA/GOx composite material was prepared according to the procedure reported in literature [5]. GOx 500 units/ml was dissolved in 1 ml aqueous solution containing 10 % PVA in ultrasonic bath for 5 min. The mixture was kept at room temperature for 6 h and then stored at −18 °C for 48 h.
3 3.1
Results End Discussion Spectroscopic Characterization
X-ray photoelectron spectroscopy was used to analyze the chemistry of PVA/GOx composite film. Figure 1 shows details of high-resolution spectra of C1s and N1s of composite material that provide binding energy information. For comparison, the sa
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