Evaluation of the activity of ß-glucosidase immobilized on polydimethylsiloxane (PDMS) with a microfluidic flow injection analyzer with embedded optical fibers.
Talanta
; 185: 53-60, 2018 Aug 01.
Article
em En
| MEDLINE
| ID: mdl-29759237
ß-glucosidase from almonds was immobilized on a polydimethylsiloxane (PDMS) microdevice by covalent chain using 3-aminopropyltrietoxysilane and glutaraldehyde. Enzymatic activity was evaluated using p-nitro-phenyl-ß-D-glucopyranoside dissolved in a 0.01â¯M pH 5.0 phosphate solution at 45⯰C measuring the reaction product (p-nitrophenol) at 410â¯nm. The microdevice consisted of two parts: the one part where the enzymatic reaction was carried out and a second part where pH was adjusted at 10, with NaOH. The reaction product was measured at the microchip exit using two optical fibers which were aligned facing each other with a gap of 7â¯mm, between both tips using guides located perpendicular to the flow outlet. A water bath was used to carry out the enzymatic reaction on the microdevice at 45⯰C. The enzymatic surface of the PDMS microdevice was 1.15â¯cm2 and the immobilized ß-glucosidase amount on the microdevice was of 1.17⯵g/cm2. The calculated kinetics parameters were: Km 2.5â¯mM; Vmax 2.2â¯mM/min; Kcat 908.3/min and Kcat/Km 363.3/mM min. The immobilized enzyme is very stable decreasing only 5% the first 15 days; on the 30th day, the activity was 69%, regarding the initial activity.
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Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Análise de Injeção de Fluxo
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Beta-Glucosidase
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Técnicas Analíticas Microfluídicas
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Dimetilpolisiloxanos
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Enzimas Imobilizadas
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Fibras Ópticas
Idioma:
En
Revista:
Talanta
Ano de publicação:
2018
Tipo de documento:
Article
País de afiliação:
México
País de publicação:
Holanda