Comparative investigation of two methods for Acetylcholinesterase enzyme immobilization on modified porous silicon

Khaldi, Khadidja; Sam, S.; Lounas, Amel; Yaddaden, C.; Gabouze, N.

doi:10.1016/j.apsusc.2016.12.169

Cited by 31 publications

(14 citation statements)

References 34 publications

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“…The contact angle measurements of the resulting PSi-AuNP-NH2 surface were found to be about 10°. This value is less than that obtained for PSi-NH2 surface (40°) [33], indicating that the presence of AuNP on the PSi surface decreased the surface hydrophobicity. The surface now bearing NH2 end groups was hence used for the covalent linking of AChE.…”

Section: Acetylcholinesterase Immobilizationcontrasting

confidence: 70%

“…The value of the oxidation peak intensity is 2.4 µA and according to the calibration curve previously established (Figure 4.b), the surface concentration of active enzymes was estimated to be Γ ≈ 1.8 x 10 12 AChE.cm -2 . This value is higher than the value obtained in our previous work [33], where the enzyme immobilization was done on SiP-NH2 surface i.e. without Au deposition.…”

Section: Enzyme Activity Assessment Of Immobilized Achecontrasting

confidence: 69%

“…Various biomonitoring methods have been developed and proposed for this purpose. Most of them rely on the colorimetric assay commonly called as the Ellman's method [31][32][33]. A few other methods, which use Fouriertransform infrared spectrometry [34], surface plasmon resonance [35] and fluorescence assay [36].…”

Section: Introductionmentioning

confidence: 99%

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Acetylcholinesterase Modified Porous Silicon for Electrochemical Measurement of Total Active Immobilized Enzyme Amount and Effective Malathion Detection

Khaldi¹,

Sam²,

Gabouze³

2020

I2M

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An amperometric method to measure the total amount of active and inhibited immobilized Acetylcholinesterase enzyme (AChE) and the quantification of AChE inhibitors was realized. In this approach porous silicon (PSi) surface was used as a matrix for AChE immobilization and Boron doped polycrystalline diamant (BDPD) was used as working electrode for the electrochemical measurements. The covalent immobilization of AChE from electric eel was achieved on amine functionalized PSi surface previously decorated with Au particles. This surface is suitable for a stable attachment of AChE enzyme. The amperometric detection of AChE activity at Boron doped polycrystalline diamond electrode is based on the oxidation of thiocholine, the enzymatic reaction product of immobilized AChE in the presence of the substrate acetylthiocholine chlorid (ATCl) and this without the need to further modify the BDPD surface or the use of other reagents. The concentration of immobilized active AChE enzymes was estimated to Γ≈ 1.8 10 12 AChE cm -2 by means of a calibration curve. Michaelis constant was assessed with a Km of 4.3 10 -4 M. Finally, the electrochemical quantification and detection of Malathion shows a good linear calibration curve where the concentration of Malathion range is from 2 to 6 nM within 6 minutes after inhibitor addition.

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Section: Acetylcholinesterase Immobilizationcontrasting

confidence: 70%

Section: Enzyme Activity Assessment Of Immobilized Achecontrasting

confidence: 69%

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Acetylcholinesterase Modified Porous Silicon for Electrochemical Measurement of Total Active Immobilized Enzyme Amount and Effective Malathion Detection

Khaldi¹,

Sam²,

Gabouze³

2020

I2M

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“…It is well known that high enzyme loading can greatly promote the catalytic efficiency of EMBRs and thus, many relevant methods have been developed in the past decades [ 9 , 13 ]. Some researchers focused on the immobilization behavior of the enzyme, including chemical bonding, physical adsorption, and entrapment [ 13 , 14 , 15 ]. Others studied ways to immobilize as many enzyme molecules as possible through surface modification of the membrane [ 9 , 16 ].…”

Section: Introductionmentioning

confidence: 99%

Immobilization of Enzymes on a Phospholipid Bionically Modified Polysulfone Gradient-Pore Membrane for the Enhanced Performance of Enzymatic Membrane Bioreactors

et al. 2018

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Enzymatic membrane bioreactors (EMBRs), with synergistic catalysis-separation performance, have increasingly been used for practical applications. Generally, the membrane properties, particularly the pore structures and interface interactions, have a significant impact on the catalytic efficiency of the EMBR. Therefore, a biomimetic interface based on a phospholipid assembled onto a polysulfone hollow-fiber membrane with perfect radial gradient pores (RGM-PSF) has been prepared in this work to construct a highly efficient and stable EMBR. On account of the special pore structure of the RGM-PSF with the apertures decreasing gradually from the inner side to the outer side, the enzyme molecules could be evenly distributed on the three-dimensional skeleton of the membrane. In addition, the supported phospholipid layer in the membrane, prepared by physical adsorption, was used for the immobilization of the enzymes, which provides sufficient linkage to prevent the enzymes from leaching but also accommodates as many enzyme molecules as possible to retain high bioactivity. The properties of the EMBR were studied by using lipase from Candida rugosa for the hydrolysis of glycerol triacetate as a model. Energy-dispersive X-ray and circular dichroism spectroscopy were employed to observe the effect of lecithin on the membrane and structure changes in the enzyme, respectively. The operational conditions were investigated to optimize the performance of the EMBR by testing substrate concentrations from 0.05 to 0.25 M, membrane fluxes from 25.5 to 350.0 L·m−2·h−1, and temperatures from 15 to 55 °C. As a result, the obtained EMBR showed a desirable performance with 42% improved enzymatic activity and 78% improved catalytic efficiency relative to the unmodified membrane.

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“…Outros trabalhos envolvendo a AChE imobilizada utilizando CLAE-EM foram descritos, como; 1) ligação covalente amino-glutaraldeído em capilar de sílica fundida [89], 2) ligação covalente amino-glutaraldeído em partículas magnéticas [94], e 3) encapsulação via sol-gel em disco monolítico [90]. Outras metodologias de imobilização para a AChE em suportes de silicone poroso com monitoramento por meio do método modificado de Ellman et al em espectrofotômetro foram publicados, sendo: 1) ligação covalente amida da enzima no suporte (silicone poroso) via processo NHS/EDC (N-hidroxisuccinimida/carbodiimida) [95]; 2) ligação covalente via reação de aminólise com grupos de ácido carboxílico da AChE e grupos amino N-terminal no suporte (silicone poroso) [95]; 3) adsorção física da enzima em silicone poroso [96]. Nestes casos, somente foram reportados o sucesso da imobilização frente a solúvel e suas possíveis aplicações.…”

Section: Métodos Com Ache Imobilizadaunclassified

Co-imobilização das enzimas acetilcolinesterase e B-secretase1: estudo de condições para triagem de ligantes

Vilela¹

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Em especial a Professora Dra. Carmen Lúcia Cardoso, pela orientação, dedicação, paciência, e direcionamento neste trabalho. Por ter me dado à oportunidade de estar em seu grupo de pesquisa, e muito mais pela confiança depositada em mim. Sou grata a você por todos estes anos de convivência e ensinamentos, que muito contribuiu para o meu crescimento científico e intelectual. Por mais que eu descreva não conseguirei te agradecer por tudo que você fez por mim, serei eternamente grata. A Olímpia Paschoal (Olimpiazinha), obrigada pelo seu auxílio, incentivo e amparo em todos os momentos, mas principalmente pela sua amizade.

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Comparative investigation of two methods for Acetylcholinesterase enzyme immobilization on modified porous silicon

Cited by 31 publications

References 34 publications

Acetylcholinesterase Modified Porous Silicon for Electrochemical Measurement of Total Active Immobilized Enzyme Amount and Effective Malathion Detection

Acetylcholinesterase Modified Porous Silicon for Electrochemical Measurement of Total Active Immobilized Enzyme Amount and Effective Malathion Detection

Immobilization of Enzymes on a Phospholipid Bionically Modified Polysulfone Gradient-Pore Membrane for the Enhanced Performance of Enzymatic Membrane Bioreactors

Co-imobilização das enzimas acetilcolinesterase e B-secretase1: estudo de condições para triagem de ligantes

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