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1.
Methods Enzymol ; 609: 1-22, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-30244787

RESUMEN

A simple method for interlocking glucose oxidase and horseradish peroxidase in a network of cellulose fibers coated with bovine serum albumin (BSA)-exfoliated graphene (biographene) is reported here. The resulting paper reactor is inexpensive and stable. Biographene is expected to function as an electron shuttle, making the reaction between the enzyme and the substrate more efficient, and this hypothesis is examined here. The BSA used to separate the sheets of graphene provides extra carboxylic acid groups and primary amines to help interlock the enzymes and the graphene in between the fibers. The decrease in entropy associated with interlocking the enzymes on a solid support is likely responsible for the increase in enzymatic stability/activity observed. Each cellulose disk contained 5.2mg of enzyme per gram of paper and 93% of the enzyme is retained after washing for 0.5-2h. This simple methodology provides a low cost, effective approach for achieving high enzymatic activity and good loadings on a benign, versatile support.


Asunto(s)
Enzimas Inmovilizadas/química , Glucosa Oxidasa/química , Grafito/química , Ingeniería de Proteínas/métodos , Celulosa/química , Peroxidasa de Rábano Silvestre , Albúmina Sérica Bovina/química
2.
Bioconjug Chem ; 29(8): 2576-2585, 2018 08 15.
Artículo en Inglés | MEDLINE | ID: mdl-29932667

RESUMEN

Synthesis, characterization, and applications of strongly fluorescent, multicolored protein nanoparticles (GlowDots) are reported here. Bovine serum albumin was cross-linked under controlled conditions to form nanoparticles, where particle size was controlled from 20 to 100 ± 10 nm by choosing appropriate reaction conditions. The absorption as well as the emission wavelengths were controlled without changing the particle size, unlike quantum dots. Each GlowDot was loaded with up to 214 ± 50 chromophores, and hence, the particles have high molar absorptivities (106 M-1 cm-1) as well as high brightness (105 to 106 M-1 cm-1). A large number of functional groups cover the particle surface and these are further functionalized to enhance cellular uptake. GlowDots that were labeled with fluorescein and functionalized with taurine, for example, were quickly taken up by HeLa, MDA-MB-231, PC3, and L6 myoblast cells, as interrogated by fluorescence imaging studies. GlowDots were biocompatible, size tunable, biodegradable, strongly fluorescent, and stable for months at room temperature, and they may serve as substitutes for quantum dots in a variety of practical applications.


Asunto(s)
Color , Nanopartículas , Albúmina Sérica Bovina/química , Línea Celular , Línea Celular Tumoral , Dicroismo Circular , Electroforesis en Gel de Agar , Electroforesis en Gel de Poliacrilamida , Colorantes Fluorescentes/química , Humanos , Microscopía Electrónica de Transmisión , Puntos Cuánticos , Albúmina Sérica Bovina/síntesis química , Albúmina Sérica Bovina/metabolismo , Espectrometría de Fluorescencia , Propiedades de Superficie
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