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

Hernández-Maya, Francisco M.; Cañizares-Macías, María P.

doi:10.1016/j.talanta.2018.03.038

Cited by 14 publications

(9 citation statements)

References 39 publications

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“…Enzyme immobilisation has proven to be an efficient tool that could retain part of the hydrolytic capability of an enzyme even for long storage periods ( Hu et al, 2018 ), so it is logical that hydrolytic activity loss is more accentuated in the free β-glucosidase than in the immobilised one. Furthermore, kinetic-constant results reported by Romo-Sánchez et al (2014) were similar to those documented in the present study whereas Hernández-Maya and Cañizares-Macías (2018) and Dal Magro et al (2019) showed that free β-glucosidase was more efficient than immobilised β-glucosidase. Catalytic efficiency may be affected after immobilisation, which could hinder the accessibility of the substrate to active sites, so the affinity of the immobilised enzyme could decrease compared with that of the free enzyme depending on the matrix and particle size used in the immobilisation procedure ( Hernández-Maya and Cañizares-Macías, 2018 ; Dal Magro et al, 2019 ).…”

Section: Discussionsupporting

confidence: 89%

Free and Immobilised β-Glucosidases in Oenology: Biotechnological Characterisation and Its Effect on Enhancement of Wine Aroma

et al. 2021

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In grapes, monoterpenes and norisoprenoids are in the form of non-volatile compounds, flavourless glycosides which could enhance the aroma of wines after its hydrolysis using β- glucosidases enzymes. It is known that the use of immobilised enzymes offers advantages such as reusability and easy recuperation. In this study, a commercial β-glucosidase was immobilised by absorption in sodium alginate. Biotechnological characteristics and terpen hydrolysis (hydrolysis aroma precursors) in muscat wines were studied after treatment with both free and immobilised commercial β- glucosidase with two different concentrations. It was revealed that both forms shared an optimal pH (4.5) and a maximum temperature (64°C), even an increment on the activity between 40and 60°C. A similar Km value has been determined while Vmax from the immobilised enzyme was higher than the free (3.35 and 2.52 μmol min–1 mg–1, respectively). Additionally, the immobilised enzyme showed a better hydrolytic activity during 24 h, and its reusability has been proven. Regarding enzymatic hydrolysis in grape must, the best results were observed for the highest concentration of free β-glucosidase although glucose release was also determined for the immobilised enzyme along the days. In contrast, maximum activity was reached by the immobilised β-glucosidase in less time but in no case equalled the free ones. Finally, volatile compound liberation in wines treated with free or immobilised enzymes was analysed using HRGC-MS. Liberation for both enzymes and the greatest concentrations of some volatiles were detected when a double dose of the free β-glucosidase was used. Nevertheless, the wines treated with the immobilised β-glucosidase showed a high concentration of some volatile compounds such as nerol or geraniol.

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Section: Discussionsupporting

confidence: 89%

Free and Immobilised β-Glucosidases in Oenology: Biotechnological Characterisation and Its Effect on Enhancement of Wine Aroma

et al. 2021

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“…It also do not hydrolyzed βGal, βXyl and αGlc glycosidic linkages. The specific enzyme activity of BglG was 5- and 303-folds higher than those of β-glucosidases from almonds and Caulobacter crescentus (Hernandez-Maya and Canizares-Macias 2018; Justo et al 2015). This result demonstrated that BglG had higher affinity to combine the substrate ρ NPG than a β-glucosidase from B. cellulosilyticus because K m of β-glucosidase from B. cellulosilyticus was 2.3-fold higher than that of BglG.…”

Section: Discussionmentioning

confidence: 95%

Identification and molecular characterization of a psychrophilic GH1 β-glucosidase from the subtropical soil microorganism Exiguobacterium sp. GXG2

Yin

et al. 2019

AMB Expr

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The products of bacterial β-glucosidases with favorable cold-adapted properties have industrial applications. A psychrophilic β-glucosidase gene named bglG from subtropical soil microorganism Exiguobacterium sp. GXG2 was isolated and characterized by function-based screening strategy. Results of multiple alignments showed that the derived protein BglG shared 45.7% identities with reviewed β-glucosidases in the UniProtKB/Swiss-Prot database. Functional characterization of the β-glucosidase BglG indicated that BglG was a 468 aa protein with a molecular weight of 53.2 kDa. The BglG showed the highest activity in pH 7.0 at 35 °C and exhibited consistently high levels of activity within low temperatures ranging from 5 to 35 °C. The BglG appeared to be a psychrophilic enzyme. The values of Km, Vmax, kcat, and kcat/Km of recombinant BglG toward ρNPG were 1.1 mM, 1.4 µg/mL/min, 12.7 s−1, and 11.5 mM/s, respectively. The specific enzyme activity of BglG was 12.14 U/mg. The metal ion of Ca2+ and Fe3+ could stimulate the activity of BglG, whereas Mn2+ inhibited the activity. The cold-adapted β-glucosidase BglG displayed remarkable biochemical properties, making it a potential candidate for future industrial applications.

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“…Micro-ow-injection analysis (mFIA) 1,2 incorporates the ow injection analysis (FIA) method on a microuidic device and is capable of high-throughput analysis because of the on-chip integration of analytical operations such as mixing, 3,4 reaction, [5][6][7] separation, 8,9 and detection. [10][11][12][13] On-chip integration is superior to conventional FIA methods as it reduces analysis time, as well as sample and reagent consumption. These advantages raise the possibility of analytical applications in diverse elds such as environmental testing, [13][14][15][16] medical diagnosis, 8,16 food analysis, 17 and drug testing.…”

Section: Introductionmentioning

confidence: 99%

Development of an on-chip sample injection system with a 6-port valve incorporated in a microchip

et al. 2020

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

Cited by 14 publications

References 39 publications

Free and Immobilised β-Glucosidases in Oenology: Biotechnological Characterisation and Its Effect on Enhancement of Wine Aroma

Free and Immobilised β-Glucosidases in Oenology: Biotechnological Characterisation and Its Effect on Enhancement of Wine Aroma

Identification and molecular characterization of a psychrophilic GH1 β-glucosidase from the subtropical soil microorganism Exiguobacterium sp. GXG2

Development of an on-chip sample injection system with a 6-port valve incorporated in a microchip

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