Evaluation of the activity of β-glucosidase immobilized on polydimethylsiloxane (PDMS) with a microfluidic flow injection analyzer with embedded optical fibers

被引:13
作者
Hernandez-Maya, Francisco M. [1 ]
Canizares-Macias, Maria P. [1 ]
机构
[1] Univ Nacl Autonoma Mexico, Dept Quim Analit, Fac Quim, Ciudad De Mexico 04510, Mexico
关键词
mu FIA; Polydimethylsiloxane enzymatic immobilization; beta-glucosidase; Enzymatic activity; Embedded optical fibers; VANILLA-PLANIFOLIA ANDREWS; ON-A-CHIP; MICROCHIP ELECTROPHORESIS; ENZYME IMMOBILIZATION; DEVICES; AGAROSE; LAB; GLUTARALDEHYDE; PURIFICATION; GLASS;
D O I
10.1016/j.talanta.2018.03.038
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
beta-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-beta-D-glucopyranoside dissolved in a 0.01 M pH 5.0 phosphate solution at 45 degrees C measuring the reaction product (p-nitrophenol) at 410 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 7 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 degrees C. The enzymatic surface of the PDMS microdevice was 1.15 cm(2) and the immobilized beta-glucosidase amount on the microdevice was of 1.17 g/cm(2). The calculated kinetics parameters were: Km 2.5 mM; Vmax 2.2 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.
引用
收藏
页码:53 / 60
页数:8
相关论文
共 49 条
[31]   INTEGRATION OF OPTOELECTRONIC COMPONENTS WITH LTCC (LOW TEMPERATURE CO-FIRED CERAMIC) MICROFLUIDIC STRUCTURE [J].
Malecha, Karol .
METROLOGY AND MEASUREMENT SYSTEMS, 2011, 18 (04) :713-721
[32]   Microfluidic lab-on-a-chip platforms: requirements, characteristics and applications [J].
Mark, Daniel ;
Haeberle, Stefan ;
Roth, Guenter ;
von Stetten, Felix ;
Zengerle, Roland .
CHEMICAL SOCIETY REVIEWS, 2010, 39 (03) :1153-1182
[33]   The effect of thermal treatment on β-glucosidase inactivation in vanilla bean (Vanilla planifolia Andrews) [J].
Marquez, Ofelia ;
Waliszewski, Krzysztof N. .
INTERNATIONAL JOURNAL OF FOOD SCIENCE AND TECHNOLOGY, 2008, 43 (11) :1993-1999
[34]   Food analysis on microchip electrophoresis: An updated review [J].
Martin, Aida ;
Vilela, Diana ;
Escarpa, Alberto .
ELECTROPHORESIS, 2012, 33 (15) :2212-2227
[35]  
Nisar A. B., 2017, INT J SYST EVOL MICR, V67, P3860
[36]   Purification and characterization of vanilla bean (Vanilla planifolia Andrews) β-D-glucosidase [J].
Odoux, E ;
Chauwin, A ;
Brillouet, JM .
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 2003, 51 (10) :3168-3173
[37]   Dual Photonic-Electrochemical Lab on a Chip for Online Simultaneous Absorbance and Amperometric Measurements [J].
Ordeig, Olga ;
Ortiz, Pedro ;
Munoz-Berbel, Xavier ;
Demming, Stefanie ;
Buettgenbach, Stephanus ;
Fernandez-Sanchez, Cesar ;
Llobera, Andreu .
ANALYTICAL CHEMISTRY, 2012, 84 (08) :3546-3553
[38]   The development and evaluation of β-glucosidase immobilized magnetic nanoparticles as recoverable biocatalysts [J].
Park, Hee Joon ;
Driscoll, Ashley J. ;
Johnson, Patrick A. .
BIOCHEMICAL ENGINEERING JOURNAL, 2018, 133 :66-73
[39]  
Patrick T., 2005, INTRO MICROFLUIDICS
[40]   A new application of aptamer: One-step purification and immobilization of enzyme from cell lysates for biocatalysis [J].
Qiao, Lifeng ;
Lv, Bo ;
Feng, Xudong ;
Li, Chun .
JOURNAL OF BIOTECHNOLOGY, 2015, 203 :68-76